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THE OHIO STATE UNIVERSITY

COLLEGE OF PHARMACY

RESEARCH DAY 2012THURSDAY, MAY 17, 2012

4-‐H CENTER

Table of Contents

WELCOME FROM THE DEAN 3

AGENDA 4

ABOUT THE SPEAKERS 5

POSTERS 6

FACULTY 6GRADUATE (MS, PHD) 7PHARMD 12POST-DOCTORAL RESEARCHERS 13RESEARCH STAFF 14RESIDENTS & FELLOWS 15UNDERGRADUATE 17

Full abstracts are available online at go.osu.edu/rd2012abstracts

The Ohio State University College of Pharmacy Research Day · May 17, 2012

Welcome from the Dean

May 17, 2012

The College of Pharmacy Research Day is an annual event held to showcase our researchactivities within our college. Research is a major enterprise of the College of Pharmacy, and covers an extensive array of areas in the pharmaceutical sciences. Faculty, graduatestudents, professional students, residents, undergraduates, and research staff areextensively involved in the exciting process of discovery and dissemination of knowledge. This year’s Research Day is one of the many activities in our College thatpromotes interdisciplinary discussions and interactions for knowledge exchange.

The College of Pharmacy Research Day 2012 includes a symposium and poster presentations of current research projects by current students and researchers. This year we are pleased to host two distinguished speakers for the symposium. Andrew Dahlem, PhD, Vice President and Chief Operating Officer, Lilly Research Laboratories, will discuss “The Future of Drug Innovation.” Horace Loh, PhD, Regents Professor, Frederick and Alice Stark Professor, and Head, Department of Pharmacology, University of Minnesota, will provide a lecture entitled “Our Search for the Ideal Analgesic in Pain Treatment.”

The depth and breadth of the research being performed in our college is outstanding. It isindeed exciting to see the results of strong disciplinary and interdisciplinary projects.

Sincerely,

Robert W. Brueggemeier, PhDProfessor and DeanThe Ohio State University College of Pharmacy


Agenda

Thursday, May 17, 201111:00 AM -‐ NOON Participant Set Up

12:30 – 2:00 Poster Presentations and Judging

2:00 – 3:00 The Future of Drug InnovationAndrew Dahlem, PhDVice President & Chief Operating OfficerLilly Research Laboratories

3:00 – 3:30 Reception

3:30 – 4:30 Our Search for the “Ideal Analgesic” in Pain TreatmentHorace Loh, PhDRegents Professor,Frederick and Alice Stark Professor, and HeadDepartment of Pharmacology, University of Minnesota

4:30 – 4:45 Poster Competition Award Presentations

4:45 – 5:00 Participants Retrieve Posters


About the Speakers

Andrew M. Dahlem, PhD, was named vice president andchief operating officer for Lilly Research Laboratories inFebruary 2007. He had been vice president of toxicology,drug disposition, pharmacokinetics, and Lilly ResearchLaboratories in Europe since January 2003 and a member ofLilly’s Senior Management Council since 2005.

Dahlem received a Bachelor of Science from The Ohio StateUniversity in 1982 and a doctorate in toxicology from theVeterinary College of the University of Illinois at Urbana-‐Champaign in 1989. He currently serves as adjunct professorof toxicology in the College of Veterinary Medicine atPurdue University, the University of Illinois at Urbana-‐Champaign, and at The Ohio State University. He is also amember of The Ohio State University College of PharmacyCorporate Council.

Horace Loh earned his PhD in biochemistry in 1965 from theUniversity of Iowa and completed a postdoctoral fellowshipin pharmacology (with Eddy Leong Way) at the University ofCalifornia-‐San Francisco. After twenty years on the faculty atUCSF Medical Center, Loh accepted the headship in theDepartment of Pharmacology at the University of Minnesotain 1989 and was named the Frederick and Alice StarkProfessor in Neuroscience. Throughout his career, he haspublished over 400 original research papers and hasmentored approximately 100 postdoctoral fellows and 30PhD students.

Loh’s laboratory continues its long-‐term investigations intothe molecular neurobiology of opioid actions and addiction.Current projects include studies on 1) the molecularmechanisms of opiate and endorphin actions; 2) themolecular mechanisms of opiate tolerance; 3) Tissue-‐specific regulation of opioid receptor gene expression; 4) Regulationof opioid receptor signal transduction; and 5) Functions ofendorphins in the CNS.


Posters

T view full abstracts, visit go.osu.edu/rd2012abstracts

Faculty

TRANSGENIC EXPRESSION OF NRF2/MAFK PROTECTS FOREBRAIN NEURONS FROM EXCITOTOXICNEURONAL DEATH IN VIVOHee-‐Yeon Cho1, Karl Obrietan2, Kari R. Hoyt11College of Pharmacy, The Ohio State University; 2College of Medicine, The Ohio State UniversityPresenting Author: Kari HoytProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 1

NOREPINEPHRINE POTENTIATING EFFECTS OF (-‐)-‐COCAINE AND RELATED TROPANE ANALOGUES ONTHE ISOLATED RAT VAS DEFERENSPopat Patil1, Richard C. Effland2, Jules B. LaPidus1,1College of Pharmacy, The Ohio State University; 2Hoechst-‐Roussel Pharmaceuticals, Inc., Somerville, NJPresenting Author: Popat PatilProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 2

RATES OF VASCULAR RELAXATION BY ANTAGONISTIC DRUGSTatiana F. González-‐Cestari1, Robert Stearns2, Popat N. Patil

1

1College of Pharmacy, The Ohio State University; 2The Ohio State University AlumnusPresenting Author: Popat PatilProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 3

MYELOPEROXIDASE (MPO)-‐DEPENDENCY FOR DNA TOPOISOMERASE II ALPHA AND BETA INHIBITION INDUCED BY THE PHENOLIC ANTICANCER AGENT ETOPOSIDE: IMPLICATIONS FOR ETOPOSIDE-‐INDUCEDLEUKEMOGENESISRagu Kanagasabai1, Sureshkumar2, Jack C. Yalowich11College of Pharmacy, The Ohio State University; 2The Ohio State University AlumnusPresenting Author: Jack YalowichProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 4


Graduate (MS, PhD)

ROLE OF TUMOR SUPPRESSIVE MICRORNAS IN PANCREATIC DUCTAL ADENOCARCINOMA.Ana Clara Azevedo-‐Pouly1, Jinmai Jiang1, Eun Joo Lee1, Vincenzo Coppola2, Gerard J. Nuovo3,David Allard4, David A. Tuveson4, Thomas D. Schmittgen11College of Pharmacy, The Ohio State University; 2Comprehensive Cancer Center, The Ohio StateUniversity; 3Phylogeny; 4Cambridge UniversityPresenting Author: Ana Clara Azevedo-‐PoulyProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical SciencesPoster Number and Abstract Page: 5

IBANDRONATE AND VENTRICULAR ARRHYTHMIA RISKIngrid M. Bonilla1, Pedro Vargas-‐Pinto1, Yoshinori Nishijima1, Adriana Pedraza-‐Toscano1, Hsiang-‐Ting Ho1,Victor Long1, Robert L Hamlin2, Sandor Gyorke1, Cynthia Carnes11College of Pharmacy, The Ohio State University; 2 Qtest Labs, Columbus, OhioPresenting Author: Ingrid BonillaProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 6

CYTOTOXICITY-‐GUIDED ISOLATION OF FLAVONOIDS AND ROTENOIDS FROM INDIGOFERA SPICATALynette Bueno Pérez1, Li Pan1, Tran Ngoc Ninh2, Hee-‐Byung Chai1, Djaja Djendoel Soejarto3, David M.Lucas4,1, A. Douglas Kinghorn11College of Pharmacy, The Ohio State University; 2Vietnamese Academy of Science and Technology,Vietnam; 3College of Pharmacy, University of Illinois at Chicago; 4College of Medicine, The Ohio StateUniversityPresenting Author: Lynette Bueno PerezProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 7

DESIGN, SYNTHESIS, AND BIOLOGICAL STUDIES OF NOVEL SURVIVIN INHIBITORSSomsundaram Chettiar1, James Cooley2, Deepak Bhasin1, In-‐Hee park1, May Mok1, Chenglong Li1, JacobNaduparambil2, Arnab Chakravarti2, Pui-‐Kai Li11 College of Pharmacy, The Ohio State University, 2 College of Medicine, The Ohio State University, Presenting Author: Somsundaram ChettiarProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 8

T315, A NOVEL INTEGRIN-‐LINKED KINASE INHIBITOR, SUPPRESSES HYPOXIA-‐INDUCEDEPITHELIAL-‐TO-‐MESENCHYMAL TRANSITION IN PROSTATE CANCERChih-‐Chien Chou, Su-‐Lin Lee, Samuel K. Kulp, Ching-‐Shih ChenCollege of Pharmacy, The Ohio State UniversityPresenting Author: Chih-‐Chien ChouProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 9


DIFFERENTIAL ANTI-‐PROLIFERATIVE ACTIVITIES OF POLY (ADP-‐RIBOSE) POLYMERASE (PARP) INHIBITORS IN TRIPLE-‐NEGATIVE BREAST CANCER CELLSHsiao-‐Ching Chuang, Naval Kapuriya, Samuel K. Kulp, Charles L. Shapiro, Ching-‐Shih Chen,College of Pharmacy, The Ohio State UniversityPresenting Author: Hsiao-‐Ching ChuangProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 10

BINDING OF HUMAN IMMUNODEFICIENCY VIRUS COFACTOR TO CHROMATINJocelyn O. Eidahl1, Brandon Crowe2, Matt Plumb1, Justin North3, Christopher McKee1, Micheal Poirier3,Mark Foster2, Mamuka Kvaratskhelia1,1College of Pharmacy, The Ohio State University; 2Department of Biochemistry, The Ohio State University; 3

Department of Biophysics, The Ohio State UniversityPresenting Author: Jocelyn EidahlProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 11

THE ROLE OF MICRORNA-‐205 IN BREAST CANCEROla Elgamal, Thomas D. Schmittgen, Jong-‐Kook Park, College of Pharmacy, The Ohio State UniversityPresenting Author: Ola ElgamalProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 12

ANTICANCER ACTIVITY AND SAR STUDIES ON VARIOUS SUBSTITUTED NAPHTHOQUINONESJonathan Etter, Deepak Bhasin, Somsundaram Chettiar, May Mok, Pui-‐Kai LiCollege of Pharmacy, The Ohio State UniversityPresenting Author: Jonathan EtterProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 13

ELECTROSPRAY ENCAPSULATION OF SUBUNIT VACCINES IN POLYMER MICROPARTICLESTony D. Duong1, Sadhana Sharma2, Kevin Peine3, Gaurav Gupta4, Abhay Satoskar4, Matthew D. Gallovic2,Eric M. Bachelder2, Barbara E. Wyslouzil1, Kristy M. Ainslie2,1College of Engineering, The Ohio State University;2College of Pharmacy, The Ohio State University;3Molecular, Cellular, and Developmental Biology, The Ohio State University; 4College of Medicine, The Ohio State UniversityPresenting Author:Matthew GallovicProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 14


THE NOVEL PROTEASOME INHIBITOR CARFILZOMIB FUNCTIONS INDEPENDENTLY OFP53 AND NF-‐ΚB TO INDUCE POTENT CYTOTOXICITY IN PRIMARY CHRONICLYMPHOCYTIC LEUKEMIA CELLSSneha Gupta1, Ellen Sass2, Erin Hertlein2, Jason Dubovsky2, Rosa Lapalombella2, Jennifer Woyach2, AmyLehman3, David Jarjoura3, John C Byrd1,2, David M Lucas1,2 ,1College of Pharmacy, The Ohio State University;2 College of Medicine, The Ohio State University;3Center for Biostatistics, The Ohio State UniversityPresenting Author: Sneha GuptaProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 15

PHARMACOKINETICS AND BLOOD-‐BRAIN BARRIER PENETRATION OF BENDAMUSTINE IN PATIENTS WITH BRAIN METASTASES FROM SOLID TUMORSLei He1, John Grecula2, Yonghua Ling1, Xiaoxia Yang1, Michael Enzerra1, Jeffrey Cotrill1, Mario Ammirati2,Kari Kendra2, Robert Cavaliere2, Nina Mayr2, John McGregor2, Thomas Olencki2, Ennio Chiocca2, JamesElder2, Mani Matharbootham2, Tim Lautenschlaeger2, Chimo Oluigbo2, Barbara McCracken-‐Bussa2, LauraMayer2, Lai Wei2, Mitch Phelphs11College of Pharmacy, The Ohio State University; 2James Cancer Hospital/OSU Comprehensive Cancer Center, The Ohio State UniversityPresenting Author: Lei HeProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 16

THE NOVEL ROLE OF INTEGRIN-‐LINKED KINASE IN IL-‐6 INDUCED TUMORIGENESISEn-‐Chi Hsu1, Yu-‐Chou Tseng1, Yo-‐Ting Tsai1, Su-‐Lin Lee1, Nicholas J. Sullivan2, Tatiana M. Oberyszyn2,Samuel K. Kulp1, Ching-‐Shih Chen11College of Pharmacy, The Ohio State University; 2Department of Pathology, The Ohio State UniversityPresenting Author: En-‐Chi HsuProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 17

OSCILLATING AQUEOUS GEL SYSTEMSTien-‐Lu Huang, Sylvan G. FrankCollege of Pharmacy, The Ohio State UniversityPresenting Author: Tien-‐Lu HuangProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 18

SYNTHESIS AND NMR ANALYSIS OF O-‐CLOSO-‐CARBORANE DERIVATIVESKeisuke Ishita, Ahmed Khalil, Rohit Tiwari, Werner TjarksCollege of Pharmacy, The Ohio State UniversityPresenting Author: Keisuke IshitaProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 19


ACTIVATION OF SILENCED TUMOR SUPPRESSOR GENES IN PROSTATE CANCER CELLSBY A NOVEL ENERGY RESTRICTION-‐MIMETIC AGENTYi-‐Chiu Kuo1, Hsiang-‐Yu Lin2,3,1, Yu-‐I Weng4, I-‐Lu Lai1, Tim H.-‐M. Huang4, Shuan-‐Pei Lin3, Dau-‐Ming Niu2,Ching-‐Shih Chen1,1College of Pharmacy, The Ohio State University; 2 Taipei Veterans General Hospital 3Mackay MemorialHospital; 4 Human Cancer Genetics Program, The Ohio State UniversityPresenting Author: Yi-‐Chiu KuoProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 20

IDENTIFICATION OF CHARAPTERIZATION OF NOVEL INTEGRIN-‐LINKED KINASEINHIBITORSu-‐Lin Lee, En-‐Chi Hsu, Chih-‐Chien Chou, Hsiao-‐Ching Chuang, Samuel K. Kulp, Ching-‐Shih ChenCollege of Pharmacy, The Ohio State UniversityPresenting Author: Su-‐Lin LeeProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 21

BIOACTIVITY-‐GUIDED ISOLATION OF ANTIOXIDANT AND QUINONE REDUCTASE-‐INDUCING CONSTITUENTS FROM ARISTOTELIA CHILENSIS (MAQUI BERRY)Jie Li1, Hee-‐byung, Chai1, Ye, Deng1, William. J., Keller2, A. Douglas, Kinghorn11College of Pharmacy, The Ohio State University; 2Nature’s Sunshine Products, Inc., Spanish Fork, UTPresenting Author: Jie LiProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 22

ANTILEISHMANIAL ACTIVITY OF A BISBENZYLISOQUINOLINE ALKALOID FROM THEROOTS OF THALICTRUM ALPINUMC. Benjamin Naman1, Gaurav Gupta2, Claudio M. Lezama-‐Davila2, Raymond W. Doskotch1, Abhay R.Satoskar2, A. Douglas Kinghorn11College of Pharmacy, The Ohio State University; 2 College of Medicine, The Ohio State UniversityPresenting Author: C. Benjamin NamanProgram of Presenting Author: Division of Medicinal Chemistry & PharmacognosyPoster Number and Abstract Page: 23

NF-‐KB-‐MEDIATED ZIP8 EXPRESSION CONTRIBUTES TO CD-‐INDUCED CELL TOXICITY ANDEMPHYSEMA IN MICEJessica Napolitano1, Mingjie Liu2, Shengying Bao2, Michael Borchers3, Daniel Nebert3,Estelle Cormet-‐Boyaka4, Patrick Nana-‐Sinkam2, Melissa Crawford2, Daren Knoell11 College of Pharmacy, The Ohio State University; 2 Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University; 3 University of Cincinnati; 4 The Ohio State UniversityPresenting Author: Jessica NapolitanoProgram of Presenting Author: Translational SciencePoster Number and Abstract Page: 24


ENDOGLIN REGULATES P38 MAPK-‐INDUCED STRESS FIBER FORMATION ANDENDOTHELIAL MIGRATIONChris Pan1, Jeff Bloodworth2, Mythreye Karthikeyan3, Arun Sharma3, Nam Lee11College of Pharmacy, The Ohio State University;2The Ohio State University; 3Duke UniversityPresenting Author: Chris PanProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 25

UNDERSTANDING THE MECHANISM OF ACTION OF ANTI-‐LEISHMANIAL ARYLIMIDAMIDE DB766Trupti Pandharkar1, Karl Werbovetz1, Frederick Buckner2, David Boykin3,1College of Pharmacy, The Ohio State University; 2University of Washington, Seattle, WA;3Georgia State University, Atlanta, GAPresenting Author: Trupti PandharkarProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 26

EFFICIENT DELIVERY OF TLR STIMULATORS TO ANTIGEN PRESENTING CELLS USINGACETYLATED DEXTRAN MICROPARTICLESKevin Peine1, Eric Bachelder1, Zachary Vangundy1, Tracy Papenfuss1, Kevin Schully2, Andrea Keane-‐Myers2,Kristy Ainslie11College of Pharmacy, The Ohio State University; 2Naval Medical Research CenterPresenting Author: Kevin PeineProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical SciencesPoster Number and Abstract Page: 27

REGULATION OF THE MACROPHAGE ZINC TRANSPORTER ZIP8 IN RESPONSE TOMYCOBACTERIUM TUBERCULOSIS INFECTIONCharlie Pyle1, Larry S. Schlesinger2, Daren L. Knoell21College of Pharmacy, The Ohio State University; 2The Ohio State UniversityPresenting Author: Charlie PyleProgram of Presenting Author: Translational SciencePoster Number and Abstract Page: 28

WATER-‐IN WATER (W/W) EMULSION SYSTEMSAnita Sharma, Sylvan G. FrankCollege of Pharmacy, The Ohio State UniversityPresenting Author: Anita SharmaProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical SciencesPoster Number and Abstract Page: 29


ALKALOIDS FROM GREWIA PANICULATA WITH CYTOTOXIC AND NON-‐COMPETITIVE NICOTINIC RECEPTOR ANTAGONISTIC ACTIVITIESPatrick Still1, Tatiana F. González-‐Cestari1, Li Pan1, Hee-‐Byung Chai1, James R. Fuchs1, Tran Ngoc Ninh2,Djaja Djendoel Soejarto3, Bitna Yi1, Brandon J. Henderson1, Popat N. Patil1, Dennis B. McKay11College of Pharmacy, The Ohio State University; 2Vietnamese Academy of Science and Technology;3University of Illinois at ChicagoPresenting Author: Patrick StillProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 30

CAVEOLIN-‐3 REGULATES SUBCELLULAR TARGETING AND FUNCTION OF KIR2.1CHANNELSZhaogang Yang, Chen Kang, Keli HuCollege of Pharmacy, The Ohio State UniversityPresenting Author: Zhaogang YangProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 31

DISCOVERY OF A NOVEL CLASS OF NEGATIVE ALLOSTERIC MODULATORS OF NACHRSAS A POTENTIAL TREATMENT FOR TOBACCO ADDICTION: STRUCTURE ACTIVITYRELATIONSHIP AND MECHANISMS OF ACTIONBitna Yi1, Tatiana F. González-‐Cestari1, Brandon J. Henderson1, Martin L. Dalefield1, Sihui Long2,Julian Richard2, Ryan Pavlovicz3, Karl Werbovetz1, Chenglong Li1, R. Thomas Boyd4, Dennis B. McKay11College of Pharmacy, The Ohio State University; 2Department of Chemistry, The Ohio State University;3Biophysics Graduate Program, The Ohio State University; 4College of Medicine, The Ohio State UniversityPresenting Author: Bitna YiProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 32

APPLICATION OF TRANSLATIONAL PK/PD MODELING TO PREDICT OPTIMAL DOSE OFSIRNA NANOPARTICLESChenguang Zhou1, Mitch A. Phelps1, L. James Lee2, Robert J. Lee11College of Pharmacy, The Ohio State University; 2College of Engineering, The Ohio State UniversityPresenting Author: Chenguang ZhouProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 33

PharmD

IMPACT OF A STUDENT PHARMACIST-‐RUN HEALTH SCREENING PROGRAM INMEDICALLY UNDERSERVED AREAS AS AN ENTRY POINT INTO THE HEALTH CARESYSTEMKyle A. Munch1, Jennifer L. Rodis1, Kristin A. Casper1, Catherine H. Kuhn2, Douglas C. Cornelius2, MichelleE. Ross11College of Pharmacy, The Ohio State University; 2The Kroger Co., Columbus DivisionPresenting Author: Kyle MunchProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 34


Post-‐Doctoral Researchers

IT TAKES MORE THAN ONE MIRNA TO TREAT HCCJon Henry, Jong-‐Kook Park, Thomas D. SchmittgenCollege of Pharmacy, The Ohio State UniversityPresenting Author: Jon HenryProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 35

SYNTHESIS OF CARBORANYL CONJUGATES OF N3-‐SUBSTITUTED THYMIDINE ANDTHEIR EVALUATION AS SUBSTRATES OF RECOMBINANT HUMAN THYMIDINE KINASE 1Ahmed Khlil1, Hitesh K. Agarwal1, Rohit Tiwari1, Kevin G. Ricks1, Elena Sjuvarsson2, Staffan Eriksson2,Michael V. Darby1, Werner Tjarks11College of Pharmacy, The Ohio State University; 2Swedish University of Agricultural SciencesPresenting Author: Ahmed KhalilProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 36

EFFECTS OF (5Z)-‐7-‐OXOZEAENOL ON PROSTATE AND BREAST CANCER CELLSUlyana Muñoz Acuña 1, Jennifer Wittwer1, Sloan Ayers2, Cedric J. Pearce3, Nicholas H. Oberlies2, Esperanza J. Carcache de Blanco11College of Pharmacy, The Ohio State University; 2The University of North Carolina at Greensboro;3Mycosynthetix, IncPresenting Author: Ulyana Muñoz AcuñaProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 37

ISOLATION, STRUCTURE ELUCIDATION, AND BIOLOGICAL EVALUATION OF 16,23-‐EPOXYCUCURBITACIN CONSTITUENTS FROM ELEAOCARPUS CHINENSISLi Pan1, Yeonjoong Yong1, Ye Deng1, Daniel D. Lantvit2, Tran Ngoc Ninh3, Heebyung Chai1, Esperanza J.Carache de Blanco1, Djaja D. Doejarto2, Steven M. Swanson2, A. Douglas Kinghorn11College of Pharmacy, The Ohio State University; 2College of Pharmacy, University of Illinois at Chicago;3Vietnamese Academy of Science and Technology, VietnamPresenting Author: Li PanProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 38

A PROSPECTIVE, DOUBLE-‐BLINDED, OBSERVATIONAL CLINICAL COHORT STUDY OF THEASSOCIATION BETWEEN TACROLIMUS EXPOSURE AND CYP3A4, CYP3A5 GENOTYPES IN ADULT HEMATOPOIETIC STEM CELL TRANSPLANT RECIPIENTSMing Poi1, Ali Mcbride2, Jigar Trivedi2, Julianna Roddy2, Jiang Wang3, Danxin Wang4, Hong Zhu5,Elizabeth Marek1, Ee Poi1, Wolfgang Sadee4, Steve Devine41College of Pharmacy, The Ohio State University; 2The Arthur G. James Cancer Hospital and Richard J.Solove Research Institute; 3The Pharmacoanalytic Shared Resource, The James Comprehensive CancerCenter; 4College of Medicine, The Ohio State University; 5College of Public Health,The Ohio StateUniversityPresenting Author:Ming J. PoiProgram of Presenting Author: Translational SciencePoster Number and Abstract Page: 39


PHARMACOKINETICS OF FLAVOPIRIDOL IN LYMPHOMASMing Poi1, Jeffrey Jones2, Amy S. Ruppert3, Jeffrey Cotrill3, Jia Ji1, Diana Chung1, Niesha Griffith4,Mitch Phelps11College of Pharmacy, The Ohio State University; 2College of Medicine, The Ohio State University; 3TheJames Comprehensive Cancer Center; 4The Arthur G. James Cancer Hospital and Richard J. SoloveResearch InstitutePresenting Author:Ming PoiProgram of Presenting Author: Translational SciencePoster Number and Abstract Page: 40

SYNTHESIS OF ELLAGIC ACID PERACETATE AND ANTITUMOR EFFICACY WITH ENHANCEMENT OF IMMUNITYYulin Ren1, Min Wei2, Patrick C. Still1, Xiaozhuo Chen3, Klaus Himmeldirk3, Michael A. Caligiuri1, A. DouglasKinghorn1, Jianhua Yu41College of Pharmacy, The Ohio State University; 2Dept. of Molecular Virology, Immunology, & MedicalGenetics, The Ohio State University; 3Ohio University; 4College of Medicine, The Ohio State UniversityPresenting Author: Yulin RenProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 41

CYTOTOXICITY, NF-‐KB P65 INHIBITION, AND IN VIVO ANTITUMOR EFFICACY OFSESQUITERPENE LACTONES FROM PIPTOCOMA RUFESCENSYulin Ren1, Ulyana Muñoz Acuña1, Daniel D. Lantvit2, Francisco Jiménez3, Ricardo García3, Heebyung Chai1,Judith C. Gallucci4, Djaja D. Soejarto2, Esperanza J. Carcache de Blanco1, Steven M. Swanson2, A. DouglasKinghorn11College of Pharmacy, The Ohio State University; 2University of Illinois at Chicago; 3Jardín BotánicoNacional “Dr. Rafael Ma. Moscoso”; 4The Ohio State UniversityPresenting Author: Yulin RenProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 42

Research Staff

IN VIVO QUANTIFICATION OF ACTIVE DECITABINE-‐TRIPHOSPHATE METABOLITE: ANOVEL PHARMACOANALYTICAL ENDPOINT FOR OPTIMIZATION OFHYPOMETHYLATING THERAPY IN ACUTE MYELOID LEUKEMIAHongyan Wang1, Ping Chen1, Jiang Wang1, Ramasamy Santhanam2, Joesephine Aimiuwu1, Vijaya Saradhi1,Zhongfa Liu1, Sebastian Schwind2, John C. Byrd2, Michael R. Grever2, Miguel A. Villalona-‐Calero2, RebeccaKlisovic2, Allison Walker2, Ramiro Garzon2, William Blum2, Kenneth K. Chan1, Guido Marucci11College of Pharmacy, The Ohio State University; 2College of Medicine, The Ohio State UniversityPresenting Author: Hongyan WangProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 43


PHARMACOKINETIC AND TISSUE DISTRIBUTION OF SYNTHETIC MIR181A AND ITS INVITRO BIOTARGETS-‐MODULATING ACTIVITYZhiliang Xie1, Hongyan Wang1, Ming Chiu1, Sebastian Schwind2, Christopher Hickey2, NatarajanMuthusamy2, Guido Marcucci1, Kenneth K. Chan11College of Pharmacy, The Ohio State University; 2Comprehensive Cancer Center, The Ohio State UniversityPresenting Author: Zhiliang XieProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 44

SIMULTANEOUS QUANTIFICATION OF 5-‐HYDROXYMETHYL-‐2’-‐DEOXYCYTIDINE ANDGLOBAL DNA METHYLATION IN DNA USING LIQUID CHROMATOGRAM TANDEM MASSSPECTROMETRYZhiliang Xie, Jiang Wang, Kenneth K. Chan, Zhongfa LiuCollege of Pharmacy, The Ohio State UniversityPresenting Author: Zhiliang XieProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical SciencesPoster Number and Abstract Page: 45

8,8-‐DIALKYLDIHYDROBERBERINES WITH POTENT ANTIPROTOZOAL ACTIVITYXiaohua Zhu1,Molla Endeshaw1, Shanshan He1, Trupti Pandharkar1, Emily Cason2, Kiran V. Mahasenan1,Chenglong Li1, Mark Bahar1, A. Douglas Kinghorn1, Mark E. Drew1, Karl Werbovetz11College of Pharmacy, The Ohio State University; 2College of Medicine, The Ohio State UniversityPresenting Author: Xiaohua ZhuProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 46

Residents & Fellows

USE OF AN ELECTRONIC MEDICAL RECORD TO IMPROVE CARE AND MONITORING OFCHRONIC KIDNEY DISEASE IN A PATIENT-‐CENTERED MEDICAL HOMEKelli Barnes1, Stuart Beatty1, Neeraj Tayal2, Amy Lehman31The Ohio State University College of Pharmacy; 2The Ohio State University College of Medicine;3BiostatisticsPresenting Author: Kelli BarnesProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 47

IMPACT OF BARCODE MEDICATION ADMINISTRATION ON EMERGENCY DEPARTMENT ERRORSJoseph Bonkowski1, Joseph Melucci1, Beth Prier1, Cynthia Carnes2, Jay Mirtallo2, Robert Weber11Wexner Medical Center at The Ohio State University; 2College of Pharmacy ,The Ohio State UniversityPresenting Author: Joseph BonkowskiProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 48


PGY-‐1 COMMUNITY CARE OSU/KROGER RESIDENTJennifer R. Frerick1, Tara R. Green21College of Pharmacy ,The Ohio State University; 2Kroger PharmacyPresenting Author: Jen FrerickProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 49

EVALUATION OF PHARMACY FACULTY KNOWLEDGE AND PERCEPTIONS OF THEPATIENT-‐CENTERED MEDICAL HOME (PCMH) CONCEPT WITHIN PHARMACYEDUCATIONAnisha Grover1, Bella H. Mehta1, Jennifer L. Rodis1, Kristin A. Casper1, Randy K. Wexler21College of Pharmacy, The Ohio State University; 2The Ohio State UniversityPresenting Author: Anisha GroverProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 50

DRUG SHORTAGES: MANAGEMENT AND RESPONSE IN HEALTH-‐SYSTEM PHARMACYDeron T. Lundy, Jennifer L. Rodis, Rodney G. Wirsching, Milap C. NahataCollege of Pharmacy, The Ohio State UniversityPresenting Author: Deron LundyProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 51

MEDICATION EVENT HUDDLES: EFFECT OF AN ELECTRONIC DATABASE ONINTERVENTION FOLLOW-‐UP IN A PEDIATRIC HOSPITALJenna Merandi1, Shelly Morvay1, Dorcas Lewe1, Barb Stewart1, Char Catt1, Jay Mirtallo2,Karl Kappeler1, Sheryl Szeinbach21Nationwide Children's Hospital; 2College of Pharmacy, The Ohio State UniversityPresenting Author: Jenna MerandiProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 52

PATIENT-‐CENTERED CARE AT A GENERAL INTERNAL MEDICINE PATIENT-‐CENTERED MEDICAL HOMEShannon Peter, Kyle Porter, Stuart J. BeattyCollege of Pharmacy, The Ohio State UniversityPresenting Author: Shannon PeterProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 53

PHARMACY RESIDENTS' PURSUIT OF ACADEMIC POSITIONSTiffany Shin, Colleen Clark Dula, Jennifer Rodis, Bella Mehta, Maria PruchnickiCollege of Pharmacy, The Ohio State UniversityPresenting Author: Tiffany ShinProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 54


MEDICATION ERRORS WITH PARENTERAL NUTRITION: IMPACT OF INGREDIENT SHORTAGESMichael Storey1, Robert Weber2, Kelly Besco2, Stuart Beatty3, Kumiko Aizawa2, Jay Mirtallo11College of Pharmacy, The Ohio State University; 2The Ohio State University; 3Ohio HealthPresenting Author:Michael StoreyProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 55

EVALUATION OF THE RATES AND CHARACTERISTICS OF ABANDONED PRESCRIPTIONS PRESCRIBED BY FEDERALLY QUALIFIED HEALTH CENTER PROVIDERS AT 340BCONTRACTED COMMUNITY PHARMACIESShannon Yarosz1, Catherine H. Kuhn21College of Pharmacy, The Ohio State University; 2Kroger Patient Care CenterPresenting Author: Shannon YaroszProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 56

Undergraduate

ZINC DEFICIENCY IN THE CONTEXT OF OBESITYEric Bolin, Shengying Bao, Daren KnoellDavis Heart and Lung Research InstitutePresenting Author: Eric BolinProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 57

TREATMENT OF AUTOIMMUNITY THROUGH TOLERANCEDeanna Brackman, Kristy Ainslie, Eric BachelderCollege of Pharmacy, The Ohio State UniversityPresenting Author: Deanna BrackmanProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 58

PURIFICATION AND IDENTIFICATION OF AN ANTILEISHMANIAL COMPOUND FROM THEROOTS OF THALICTRUM RUGOSUMAnthony Gromovsky1, C. Benjamin Naman1, Gaurav Gupta2, Claudio M. Lezama-‐Davila2, Raymond W.Doskotch1, Abhay R. Satoskar2, A. Douglas Kinghorn11College of Pharmacy, The Ohio State University; 2College of Medicine, The Ohio State UniversityPresenting Author: Anthony GromovskyProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 59


SYNTHESIS AND CHARACTERIZATION OF ETHANOL-‐DEGRADING ACETALATEDDEXTRAN POLYMER AND MICROPARTICLESKevin J. Kauffman1, Clement Do2, Sadhana Sharma2, Eric M. Bachelder2, Kristy M. Ainslie21College of Engineering, The Ohio State University; 2College of Pharmacy, The Ohio State UniversityPresenting Author: Kevin KauffmanProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 60

ANTIPSYCHOTIC DRUG IMPACT ON DOPAMINERGIC NEURONSAlex KlausingCollege of Pharmacy, The Ohio State UniversityPresenting Author: Alex KlausingProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 61

DEPLETION OF PIN1 IN MOUSE AORTIC ENDOTHELIAL CELLS INCREASES INDUCTION OF HYPOXIA-‐INDUCIBLE TRANSCRIPTION FACTORSAbir Mneimneh, Dale Hoyt, College of Pharmacy, The Ohio State UniversityPresenting Author: Abir MneimnehProgram of Presenting Author: Division of PharmacologyPoster Number and Abstract Page: 62

IMMUNOSTIMULATORY POLYSACCHARIDES AS A BASE MATERIAL FOR POLYMERIC MICROPARTICLESDoug Montjoy1, Kevin Peine2, Eric M. Bachelder1, Kristy M. Ainslie11College of Pharmacy, The Ohio State University; 2Molecular Cell and Developmental Biology, The Ohio State UniversityPresenting Author: Doug MontjoyProgram of Presenting Author: Division of Pharmaceutics and Pharmaceutical ChemistryPoster Number and Abstract Page: 63

EVALUATING DRUG THERAPY DECISION MAKING IN PATIENTS WITH EPILEPSYJames McAuley1, Abby M. Stazar1, Yun Lee1, Sheri Cotterman-‐Hart2, Bassel Shneker21College of Pharmacy, The Ohio State University; 2College of Medicine, The Ohio State UniversityPresenting Author: Abbey StrazarProgram of Presenting Author: Pharmacy Practice and AdministrationPoster Number and Abstract Page: 64

SYNTHESIS OF ALLOSTERIC MODULATORS FOR NICOTINIC ACETYLCHOLINE RECEPTORSJason Young, Bitna Yi, Tatiana Gonzalez-‐Muskus, Dennis McKay, Karl Werbovetz, College of Pharmacy, The Ohio State UniversityPresenting Author: Jason YoungProgram of Presenting Author: Division of Medicinal Chemistry and PharmacognosyPoster Number and Abstract Page: 65


TRANSGENIC EXPRESSION OF NRF2/MAFK PROTECTS FOREBRAINNEURONS FROM EXCITOTOXIC NEURONAL DEATH IN VIVO

Hee-‐Yeon Cho, Karl Obrietan, Kari R. HoytOSU College of Pharmacy, Division of Pharmacology and OSU College of Medicine,Department of Neuroscience

Rank of last author: FacultyProgram of last author: Division of Pharmacology

We are examining whether upregulation of neuronal antioxidant response element(ARE) regulated gene expression is neuroprotective in the setting of excitotoxic braininjury. To achieve this goal, we are using a novel transgenic mouse strain that wasengineered to express the ARE transcription factors Nrf2 and MafK (and the cell markereGFP) in neurons in a tetracycline-‐regulatable manner. We first verified that Nrf2 andgst-‐1 (a phase II target gene) mRNA expression levels were increased in Nrf2/MafKtransgenic mouse brain tissue. To test for a potential neuroprotective role of neuronalNrf2/MafK expression, we used pilocarpine-‐induced status epilepticus (SE) to induceexcitotoxic damage, and assessed neuronal death and markers of Nrf2 activation 8 and72 hours post SE. We observed robust SE-‐induced cell death (measured by fluoro-‐jade Bstaining and calpain activation) which was significantly attenuated in Nrf2/MafKtransgenic mice compared to wildtype (WT) mice. In Nrf2/MafK transgenic mice, Nrf2 strongly translocated to the nucleus in hippocampal and striatal neurons afterexcitotoxic stimulation. Induction of the Nrf2-‐dependent antioxidant gene, HO-‐1, wasassessed by immunoblotting and immunofluorescence labelling in parallel with the celldeath assessments. To specifically assess neuronal HO-‐1 expression, we double-‐labelled brain tissue with HO-‐1 and NeuN (a neuronal marker) antibodies. The number of HO-‐1/NeuN positive cells was 2.5 fold higher in Nrf2/MafK transgenic mice than in WTlittermates after SE. In summary, we have established that neuronal Nrf2/MafKupregulation and concomitant detoxifying/antioxidant induction is a viable therapeutictarget for the design of therapies to protect neurons from excitotoxic/oxidative injury.

The Ohio State University College of Pharmacy Research Day Abstract Page and Poster Number 1

NOREPINEPHRINE POTENTIATING EFFECTS OF (-‐)-‐COCAINE AND ITSANALOGUES RELATED TROPANE ANALOGUES ON THE ISOLATED RAT VASDEFERENS*

Popat N. Patil, Richard C. Effland1, Jules B. LaPidusCollege of Pharmacy, The Ohio State University, Emeritus Professor, Pharmacology1Hoechst-‐Roussel Pharmaceuticals, Inc., Somerville, NJ, Director (retired), MedicinalChemistry

Rank of first author: FacultyProgram of first author: Division of Pharmacology

The natural productΨ-‐tropine analogue, (-‐)-‐Cocaine, is well known for itsnorepinephrine potentiating action by inhibiting neuronal uptake at the adrenergicsynapse. Related tropine analogue atropine, a potent antimuscarinic, lacks this effect.Actions of tropine andΨ-‐tropine analogues were examined to understand the stericstructural requirements for the potentiating effect. Rat vas deferens was used as it isvery sensitive to drug potentiation. The organ was mounted in the oxygenated tissuebath containing physiological salt solution at 37.5°C. The Control CumulativeConcentration response of the organ to the norepinephrine or the indirectly actingtyramine was obtained. After thorough wash, the tissue was incubated with the drugfor 3 minutes, and norepinephrine or tyramine responses were observed. Theconcentration of the drug that caused ~3-‐fold shift of the norepinephrine curve to theleft was used to compare relative potentiation by analogues. The relative potentiationby drugs were: (-‐)-‐Cocaine 1, Tropine benzoate, 1/30th, (+)-‐Ψ-‐Cocaine, (±)Allo-‐Ψ-‐Cocaine, andΨ-‐Tropine benzoate, 1/100th, and Benzoylecgonine,Ψ-‐Ecgonine methylester, (±)-‐Allo-‐Ψ-‐Ecogonine methyl ester, Tropine andΨ-‐Tropine, 1/1000th. (-‐)-‐Cocaine at 10-‐6M caused equal shift of the norepinephrine and tyramine dose response curves tothe left and right, respectively. Tropine benzoate at 3x10-‐5M shifted the curve ofnorepinephrine by ~3-‐fold to the left did not change the responses to tyramine. Ψ-‐Tropine benzoate at 10-‐4M also potentiated norepinephrine by ~3-‐fold but markedlyinhibited responses to tyramine. Thus these two analogues dissociated the actions ofnorepinephrine and tyramine (at the transport) in contrast to (-‐)-‐cocaine which impactsboth actions. Relationship of reactive groups of these molecules to that of thefunctional effects will be presented.

*In part from the 1971 Ph.D. thesis of R. Effland, is a tribute to our late Jules LaPidus, Professor ofMedicinal Chemistry, Dean of Graduate School, who over 40 years ago stimulated our interest in the stericaspects of drug action.


RATES OF VASCULAR RELAXATION BY ANTAGONISTIC DRUGSTatiana F. González-‐Cestari, Robert Stearns, Popat N. PatilDivision of Pharmacology, College of Pharmacy, The Ohio State University.

Rank of last author: FacultyProgram of last author: Division of Pharmacology

Majority of mammalian blood vessels are innervated by post ganglionic sympatheticnerves. The activation of the neuron releases (-‐)-‐norepinephrine and α-‐adrenoceptor mediated vasoconstriction is produced. Blockade of this receptor causes vascularrelaxation. Many pharmacologic agents as well as endogenous substances producevascular relaxation by different mechanisms. On the isolated rat aorta, we haveinvestigated the rates of relaxation by the different drugs of phenylephrine-‐ oroxymetazoline-‐mediated vasospasm. The project was initiated with the idea thatcomparative relaxation rates should explain the therapeutic rationale for the utility ofdrugs in vasospastic diseases. In vitro, the α-‐adrenoceptor activators phenylephrine oroxymetazoline, at approximately EC50 concentration, were used to produce a stablecontraction of the aorta and, at the response plateau, a concentration of the musclerelaxant drug was added. Rate of vascular relaxation was observed. A number of 3-‐7 experiments were repeated and data were averaged for calculation of time required forhalf the initial contraction of the aorta (T1/2 values). Blockade of phenylephrine-‐induced vasospasm with a concentration of phentolamine (100 nM), a competitive reversible α-‐adrenoceptor antagonist, occurred with a T1/2 value of 1.8 min. On the other hand, theirreversible α-‐adrenoceptor antagonist, dibenamine, caused slow relaxation in thepresence of phenylephrine, with T1/2 = 11.8 min. The muscarinic cholinergic agonists,carbachol, pilocarpine, and arecoline showed quick relaxation with T1/2 values of 0.3,0.3, and 0.4 min, respectively. The calcium channel antagonist, nifedipine, andphosphodiesterase inhibitors, theophylline and papaverine, exhibited relaxation withT1/2 values of 1.8, 0.6, and 2.2 min, respectively. Our results indicate that depending onthe mechanism of action of the antagonistic drug, T1/2 values for the vascular relaxationvary greatly. Equilibrium studies of vascular relaxation can compromise the importanceof one drug over the other in understanding vasospastic mechanisms.


MYELOPEROXIDASE (MPO)-‐DEPENDENCY FOR DNA TOPOISOMERASE IIALPHA AND BETA INHIBITION INDUCED BY THE PHENOLIC ANTICANCERAGENT ETOPOSIDE: IMPLICATIONS FOR ETOPOSIDE-‐INDUCEDLEUKEMOGENESIS.

Ragu Kanagasabai, Ph.D., Sureshkumar, M.A.,Ph.D., Jack C.Yalowich, Ph.D.Division of Pharmacology, College of Pharmacy

Rank of first author: Research StaffProgram of first author: Division of Pharmacology

The clinical efficacy of the anticancer agent etoposide (VP-‐16) is compromised by itsability to cause acute myeloid leukemias (t-‐AML) associated with MLL gene rearrangements. We proposed previously that myeloperoxidase (MPO) found in myeloidprecursors converts the phenolic drug VP-‐16 to its phenoxyl radical (VP-‐16-‐O·∙), whichredox cycles, leading to the generation of reactive oxygen species, oxidative DNA damage linked to DNA topoisomerase II (topo II)-‐mediated strand cleavage, andresultant recombination events causal for t-‐AML. In the present study we have utilizedan shRNA for MPO in myeloid leukemia HL-‐60 cells to demonstrate MPO dependencyfor a portion of VP-‐16 inhibition/poisoning of topo II isoforms. HL-‐60 cells wereinfected with lentivirus containing shRNA for GFP or MPO. All infected cell lines grewwith similar doubling times (17-‐19 hr) and contained comparable levels of topo II alphaand beta, and DNA topoisomerase I (topo I). In MPO knockdown cells, mature MPOexpression was reduced to 6% of the level found in GFP shRNA controls. MPO activity incell lysates from MPO shRNA infected cells was reduced to 15% of controls. In HL-‐60 control cells (GFP shRNA), VP-‐16 (0-‐100 µM) induced a progressive increase in DNA double strand breaks assessed by levels of phosphorylated gamma-‐H2AX. In contrast,VP-‐16-‐induced DNA breakage was attenuated in MPO-‐knockdown cells (shRNA MPO).Using immunoblots of cell lysates, topoisomerase band depletion studies revealed thatVP-‐16-‐induced topo II alpha-‐ and topo II beta-‐DNA covalent complexes were decreasedin MPO knockdown compared to GFP control cells. VP-‐16 had no effect on topo I-‐DNAcomplexes. In separate studies, direct topo II isoform complexes with genomic DNA were analyzed in GFP shRNA and MPO shRNA infected cells. Using this complementarytechnique, VP-‐16-‐induced topo II-‐DNA complexes were reduced in MPO knockdowncells compared to shRNA GFP controls. Overall, these new studies, along withpreviously published work (Mol. Pharmacol. 79(3): 479-‐497, 2011), implicate MPO-‐mediated oxidative activation of VP-‐16 in production of genotoxic and leukemogeniceffects known to derive secondary myeloid leukemias after patient treatment with thisagent.


ROLE OF TUMOR SUPPRESSIVE microRNAS IN PANCREATIC DUCTAL ADENOCARCINOMA

Ana Clara Azevedo-‐Pouly1, Jinmai Jiang1, Eun Joo Lee1, Vincenzo Coppola2, Gerard J.Nuovo3, David Allard4, David A. Tuveson4, and Thomas D. Schmittgen11College of Pharmacy, Ohio State University, Columbus, OH; 2Comprehensive Cancer Center, Ohio State University, Columbus, OH; 3Phylogeny, Columbus, OH; 4CambridgeUniversity, Cambridge, UK

Rank of first author: Graduate StudentProgram of first author: Division of Pharmaceutics and Pharmaceutical Sciences

microRNAs (miRNAs) are differentially expressed in many cancers including pancreaticductal adenocarcinoma (PDAC). We performed qPCR profiling studies of primary andmature miRNA in PDAC specimens from patients, pancreatic cancer cell lines and in thepancreas of transgenic KRASG12D transgenic mice. Major deregulation of the miRNAs was seen in the PDAC tumors with 115 of 174 expressed miRNAs (66%) increased in thetumors (P < 0.05). Only 5 miRNAs had a pattern of down-‐regulation from normal totumor, including the pancreas specific miR-‐216a, miR-‐216b and miR-‐217. Results werecorroborated in tissue from two different KRASG12D transgenic mouse models. Tissuespecific miRNAs are down-‐regulated in various cancer types and reverse the malignantphenotype when miRNA mimic oligos are re-‐introduced into cancer cell lines. In situhybridization of miR-‐216/-‐217 in adult, human pancreas showed that it is expressedmost abundantly in acini. We hypothesize that this down-‐regulated cluster of miRNAs (miR-‐216a, miR-‐216b and miR-‐217) targets epithelial genes and might serve as agatekeeper for the acinar-‐to-‐ductal metaplasia (ADM) seen in PDAC. A three-‐dimensional ADM cell culture model was employed by culturing primary mouse acini onmatrigel. Ductal differentiation, (marked by increasing levels of epithelial markers,decreased levels of acinar markers and duct formation), showed decreasing levels ofmiR-‐216a, miR-‐216b and miR-‐217. Transfection of a miR-‐217 oligo mimic into pancreaticcell lines led to a down regulation of KRAS and CDH1 proteins. Epigenetic regulation ofthe cluster was studied by treating PDAC and pancreas cell lines with 5-‐aza 2'deoxycytidine and Trichostatin A. Current results show that the mechanism ofregulation of these miRNAs does not depend on miRNA biogenesis but might rely onhistone methylation. The ability of miR-‐216a, miR-‐216b and miR-‐217 to induce ADMand possibly PDAC will be studied in a miRNA knockout mouse that is currently underdevelopment.

Financial Support: NCI grant 2U01CA111294-‐06.


IBANDRONATE AND VENTRICULAR ARRHYTHMIA RISK

Ingrid M Bonilla, Pedro Vargas-‐Pinto, Yoshinori Nishijima,Adriana Pedraza-‐Toscano, Hsiang-‐Ting Ho,Victor Long, Andriy Belevych, Chun Li, Jessica Smith, Mahmoud Houmsse, Troy Rhodes, Raul Weiss, OHIO STATE UNIV, Columbus,OH; Robert L Hamlin, Qtest Labs, Columbus, OH; Sandor Gyorke, Cynthia ACarnes, OHIO STATE UNIV, Columbus, OH

Rank of First Author: Graduate StudentProgram of First author: Division of Pharmacology

Abstract:BACKGROUND: Ibandronate is given monthly or quarterly to post-‐menopausalwomen to treat or reduce osteoporosis. We report a case of cardiac arrestin an otherwise healthy 55 year old female, 2 weeks after her first dose ofibandronate, with associated hypokalemia and QTc prolongation (575 ms). TheQT/QTc remained prolonged after correction of hypokalemia; the QT/QTcreturned toward normal after drug discontinuation.METHODS: A single 3 mg injection was given to four dogs; ECG was monitoredfor six weeks by telemetry. Langendorff perfused guinea pig hearts were usedto evaluate acute ibandronate effects on repolarization. Adult canineventricular myocytes were used to study action potentials (0.5, 1, and 2Hz), potassium currents and spontaneous calcium waves during treatment withibandronate (0.001 -‐ 1000 mcg/L), and washout.RESULTSs: Three of 4 dogs exhibited increased short-‐term variabilityin repolarization only days after the dose. There was noevidence of QT/QTc changes in the Langendorff experiments. In myocytes,ibandronate exposure caused reverse-‐use dependent prolongation of the actionpotential (APD50 and APD90), as well as increased beat-‐to-‐beat variability in APD90(p<0.05). Early and delayed afterdepolarizations were observed in ~45% of myocyteswith ibandronate and in 33% during washout. Notably, ibandronate did not alter Ito,IK1, IKr or IKs. In separate experiments, spontaneous intracellular calcium waves wereincreased only during washout of ibandronate (p<0.05). Buffering calcium with BAPTAprevented ibandronate-‐induced APD90 instability and afterdepolarizations.CONCLUSIONS: Ibandronate alters ventricular myocyte electrophysiology and calciumhandling to induce cellular arrhythmias; in vivo data is consistent with late onset oftorsadogenic repolarization instability. Collectively, these data suggest thatabnormalities in myocyte calcium regulation underlie the observed arrhythmias. Weconclude that current paradigms to detect proarrhythmia risk may not be sufficientlysensitive.


CYTOTOXICITY-‐GUIDED ISOLATION OF FLAVONOIDS AND ROTENOIDSFROM INDIGOFERA SPICATA

Lynette Bueno Pérez 1, Li Pan1, Tran Ngoc Ninh 2, Hee-‐Byung Chai1, Djaja Djendoel Soejarto 3,David M. Lucas1,4, and A. Douglas Kinghorn11 Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio State University,Columbus, OH 43210, USA; 2Institute of Ecology and Biological Resources, Vietnamese Academy ofScience and Technology, Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam;3Program for Collaborative Research in the Pharmaceutical Sciences, Department of Medicinal Chemistryand Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, Chicago, IL 60612, USA; 4Department of Internal Medicine, College of Medicine, The Ohio State University, Columbus, OH 43210,USA

Rank of first author: Graduate Student Program of first author: Division of Medicinal Chemistry and Pharmacognosy

Indigofera spicata Forssk. (Leguminosae) is native to parts of East Africa, Madagascar,the Philippines, and Indonesia. However, I. spicata has become widespread in tropicaland sub-‐tropical countries as a ground cover plant.1 The bioactivity-‐guided fractionationof a cytotoxic chloroform extract of the flowers, fruits, leaves, and twigs of I. spicata,with IC50 values in HT-‐29 cells of 4.3 μg/mL, in 697 cells of 2.2 μg/mL, and in Raji cells of5.0 μg/mL, was performed, leading to the isolation of five compounds (1-‐5). A semi-‐synthetic compound, (+)-‐tephrosone (6) was produced by alkaline hydrolysis of (+)-‐purpurin (1). The structures of these compounds were established on the basis ofspectroscopic methods, including NMR and HR-‐MS, as well as by comparison of theirspectroscopic data with those reported in the literature. The structure of the newcompound 2 was determined by 1D-‐, 2D-‐NMR and HR-‐MS. The relative configuration ofcompound 2 was determined by measuring its optical rotation and CD spectrum, andcomparing these data to those of compound 1. Compound 4 exhibited potentcytotoxicity for HT-‐29 and 697 cells, with IC50 values of less than 1 μM. Compound 5showed cytotoxicity for 697 cells but was inactive against HT-‐29 and Raji cells.Compounds 1-‐3, and 6 were inactive for all the cell lines tested. This represents the firstreport of the isolation of compounds 1-‐5 from I. spicata. Also, a new compound, (+)-‐10-‐deacetyl-‐purpurin (2) was isolated from this plant. The cytotoxicity of compound 4 hasbeen reported previously using other cancer cell lines but this is the first report of suchinhibitory activities for HT-‐29, 697, and Raji cells. No in vivo studies of compound 4 havebeen performed so far. This represents a potential candidate lead compound for furtherstudies as a possible anticancer agent.

1. Morton, J. F. Econ. Bot. 1989, 69, 314-‐327.

Mr. Mark Apsega from the Mass Spectrometry & Proteomics Facility at The Ohio State University (OSU) isacknowledged for training and help with the use of the MS instrument. In addition, Dr. Chunhua Yuan isacknowledged for running compounds 2 and 4 in the 800 MHz NMR spectrometer at the OSU CampusChemical Instrument Center, NMR facility. Support from grant P01 CA125066 from the National Cancer

Institute, NIH, Bethesda, MD, is also acknowledged.


DESIGN, SYNTHESIS, AND BIOLOGICAL STUDIES OF NOVEL SURVIVININHIBITORSSomsundaram Chettiar,1 James Cooley,2 Deepak Bhasin,1 In-‐Hee park,1May Mok,1

Chenglong Li,1 Jacob Naduparambil,2 Arnab Chakravarti2 and Pui-‐Kai Li11Division of Medicinal chemistry & Pharmacognosy, College of Pharmacy, The Ohio StateUniversity, Columbus, Ohio, 432102Department of Radiation Oncology, College of Medicine, The Ohio State University,Columbus, Ohio, 43210

Rank of first author: Graduate StudentProgram of first author: Division of Medicinal Chemistry and Pharmacognosy

Survivin, a member of inhibitors of apoptosis (IAP) protein family is involved in celldivision and has anti-‐apoptotic activity. Survivin is undetectable in non-‐proliferating cells but is over-‐expressed in various cancers. Over-‐expression of survivin in cancerpatients has been associated with poor patient survival rates, metastasis, increased recurrence and resistance to radiation therapy and chemotherapy.1 Thus, survivin is oneof the viable molecular targets for the treatment of cancer.

Recently, through HTS-‐NMR and AS/MS affinity based screenings, Abbott8 wasidentified to bind to the dimerization interface of survivin.2 Structure based designapproach was used with Abbott8 as the lead structure to design several small moleculeinhibitors. The time and dose dependant effects of LLP compounds were compared bytracking green fluorescence signal from GFP-‐tagged Survivin associated withChromosomal Passenger Complex (CPC) in HUVEC and PC3cells. Two of the designedinhibitors, LLP3 and LLP9 caused delay in mitosis and imparted major defects in CPCorganization and mitotic progression at 50 nM and 100 nM, respectively. Structure-‐activity-‐relationship studies will be presented in detail.

References:

1. Pennati, M.; Folini, M.; Zaffaroni, N., Targeting survivin in cancer therapy. Expert Opin Ther Targets2008, 12, 463-‐476.2.Wendt, M. D.; Sun, C.; Kunzer, A.; Sauer, D.; Sarris, K.; Hoff, E.; Yu, L.; Nettesheim, D. G.; Chen, J.; Jin, S.; Comess, K. M.; Fan, Y.; Anderson, S. N.; Isaac, B.; Olejniczak, E. T.; Hajduk, P. J.; Rosenberg, S. H.; Elmore,S. W., Discovery of a novel small molecule binding site of human survivin. Bioorg Med Chem Lett 2007, 17,3122-‐3129.


T315, A NOVEL INTEGRIN-‐LINKED KINASE INHIBITOR, SUPPRESSESHYPOXIA-‐INDUCED EPITHELIAL-‐TO-‐MESENCHYMAL TRANSITION INPROSTATE CANCER

Chih-‐Chien Chou, Su-‐Lin Lee, Samuel K. Kulp, Ching-‐Shih ChenDivision of Medicinal Chemistry, College of Pharmacy, The Ohio State University


Objectives of the study:The epithelial-‐to-‐mesenchymal transition (EMT) is an early event in metastasisthat involves the loss by epithelial cells of many of their distinctive epithelial characteristics and theacquisition of mesenchymal properties. EMT can be induced in cancer cells by several factors, such ashypoxia and TGF-‐β1, leading to an aggressive and malignant phenotype. In prostate cancer therapy, EMTcan be a major clinical challenge as it can contribute to tumor recurrence, therapy resistance, andmetastasis. Recently, the integrin-‐linked kinase (ILK) has been identified as an important protein involvedin the process of EMT by inducing the protein expression and activation of Snail, one of the major EMTmarkers in various cancer cells. The objective of this study was to evaluate the ability of T315, a novel ILKinhibitor developed in our laboratory, to block hypoxia-‐induced EMT in prostate cancer cells and tovalidate the role of ILK in hypoxia-‐induced EMT.Methods Used:Western blot analysis was used to assess the effect of EMT inducer (hypoxia and TGF-‐β1)and T315 on the expression levels epithelial marker, such as E-‐cadherin and mesenchymal markers including, vimentin, Snail, and Zeb1, in PC-‐3 cells. Cells were exposed to hypoxic condition (0.5% O2) in ahypoxia chamber (BioSpherix). To clarify that the effect of T315 on EMT was ILK-‐dependent, stabletranfectants expressing constitutively active (CA)-‐ILK were used. Immunofluorescence was used toexamine the expression and localization of EMT markers. In order to observe the effect of T315 on themotility of PC-‐3 cells after hypoxia-‐induced EMT, the Transwell cell migration assay and wound healingmigration assay were used. To examine the transcriptional regulation of Snail by HIF1α, luciferasereporter assay and RT-‐PCR were used to examine Snail promoter activity and mRNA level of Snail in PC-‐3 cells, respectively. To assess the effect of T315 on Snail phosphorylation and stability, western blottingand immunoprecipitation analysis were used.Results and Conclusion: Based on the results of western blot analysis and immunofluorescence, EMT wasinduced by hypoxia and TGF-‐β1 treatment in PC-‐3 cells. Moreover, T315 was shown to counteract theeffects of hypoxia on EMT markers by efficiently increasing the protein expression level of E-‐cadherin anddecreasing those of HIF1α, vimentin, Snail, and Zeb1 in PC-‐3 cells. Besides, F-‐actin immunofluorescence staining also showed the transition of epithelial phenotype to mesenchymal phenotype under hypoxic condition and this phenomenon was reversed by T315 treatment. More importantly, in the Transwell and wound healing migration assays, T315 showed its ability to inhibit hypoxia-‐induced cell motility in a dose-‐dependent manner. Western blotting results showed that T315 downregulated PKB/Akt and mTORactivities and decreased HIF1α expression. Furthermore, luciferase report assay revealed that T315 coulddownregulate Snail promoter activity induced by HIF1α. According to a previous study, GSK3β-‐mediatedSnail phosphorylation altered the nuclear sequestration of Snail and caused Snail to undergo proteasomal degradation. Our western blotting and immunoprecipitation results showed that T315 could cause GSK3β dephosphorylation, increase the phosphorylation status of Snail and promote Snail degradation.Together, these findings suggest that T315 can inhibit hypoxia-‐induced EMT in PC-‐3 cells and that this ismediated through the AKT/mTOR/HIF1α pathway.Significance: These results indicate that the inhibition of ILK can block hypoxia-‐induced EMT in prostatecancer cells as reflected by changes in molecular markers and cell behavior, and that this inhibition can beachieved by treatment with the novel small molecule agent T315, which may have therapeutic benefitsfor prostate cancer patients with subsequent metastasis.Acknowledgement: We thank Su-‐Lin Lee for T315 synthesis.


DIFFERENTIAL ANTI-‐PROLIFERATIVE ACTIVITIES OF POLY (ADP-‐RIBOSE)POLYMERASE (PARP) INHIBITORS IN TRIPLE-‐NEGATIVE BREAST CANCERCELLS

Hsiao-‐Ching Chuang, Naval Kapuriya, Samuel K. Kulp, Charles L. Shapiro, and Ching-‐Shih ChenDivision of Medicinal Chemistry, College of Pharmacy, The Ohio State University (OSU)


Objectives of the Study: Triple-‐negative (ER-‐negative, PR-‐negative, HER2/neu-‐negative) breast cancer (TNBC) is a clinical challenge because of the lack of a specific therapeutictarget. Poly (ADP-‐ribose) polymerase (PARP) is a nuclear enzyme involved in thedetection and repair of DNA damage. In DNA repair-‐defective tumors inhibition of PARPcan cause genomic instability and cell death. Currently, four commercial PARP inhibitors[AZD-‐2281, AG-‐014699, ABT-‐888 and BSI-‐201] are in clinical trials in TNBC. Recent results of a randomized phase II clinical trial showed that BSI-‐201 when added toplatinum-‐containing combination chemotherapy statistically significantly improved theoutcome of metastatic breast cancer patients relative to chemotherapy alone. Despiterecent advances in the clinical evaluation of various PARP inhibitors in TNBC patients, invitro data to define potential anti-‐tumor mechanisms beyond PARP inhibition for theseagents are lacking.Methods: MDA-‐MB-‐468, MDA-‐MB-‐231 and Cal-‐51 cells were obtained from ATCC andDSMZ and maintained in complete growth medium. The four PARP inhibitors, AZD-‐2281, AG-‐014699, ABT-‐888 and BSI-‐201 were evaluated in cell survival assays [MTT andclonogenic formation]. PARP-‐dependent and-‐independent signaling mechanisms of eachPARP inhibitors were investigated by Western blotting and shRNA-‐mediated knockdown. Potential synergistic interactions between PARP inhibitors and cisplatin insuppressing TNBC cell viability were assessed.Results: The four PARP inhibitors exhibited differential antitumor activities, with therelative potencies of AG-‐014699 > AZD-‐2281 > ABT-‐888 > BSI-‐201. The higher potenciesof AG-‐ 014699 and AZD-‐2281 were associated with their effects on G2/M arrest andDNA damage as manifested by γ-‐H2AX formation. Abilities of individual PARP inhibitorsto sensitize TNBC cells to cisplatin varied to a great extent in a cell context-‐ and cell line-‐specific manner.Significance: These studies will contribute to a deeper understanding of potentialdifferences in the PARP inhibitors which is essential for planning preclinical studies ofthe PARP inhibitors as single agents and in combination with other novel therapeutics.


BINDING OF HUMAN IMMUNODEFICIENCY VIRUS COFACTOR TOCHROMATIN

Jocelyn O. Eidahl1, Brandon L. Crowe2, Justin North3, Matt Plumb1, Christopher M.Mckee1, Micheal G. Poirier3, Mark P. Foster2, Mamuka Kvaratskhelia1

OSU College of Pharmacy1, OSU Department of Biochemistry2, OSU Department ofBiophysics3

Rank of first author Graduat StudentProgram o first author Division of Pharmaceutics and Pharmaceutical Sciences

HIV-‐1 replication requires integration of the viral cDNA made by reverse transcription into the host chromosome for viral replication and maintenance of the infected state in the host. Theintegration step is catalyzed by the retroviral enzyme integrase (IN). In vivo this process isregulated by multiple viral and cellular cofactors. Among these, lens epithelium-‐derived growth factor (LEDGF) has been identified as the principal cellular cofactor essential for effective HIV-‐1 integration. LEDGF tethers the HIV-‐1 IN to active transcription units in the host chromatin. The C-‐terminal region of LEDGF contains the integrase binding domain, IBD, which binds HIV-‐1 IN.While the N-‐terminal domain ensemble of LEDGF is essential for chromatin binding, themolecular mechanism of these interactions is unknown. A component of the N-‐terminal domainensemble, the PWWP domain, is necessary for interactions to chromatin. Deletion of the PWWP domain causes reduced HIV integration into active transcription units. PWWP domains can befound in over 60 eukaryotic proteins. Of the PWWP containing proteins that have been studied,results show these domains to contain a hydrophobic cavity that recognizes specific core histonepost-‐translational modifications. We hypothesize the PWWP domain of LEDGF recognizes aspecific core histone post-‐translational modification while simultaneously binding HIV-‐1 INthrough its IBD allowing HIV-‐1 integration to occur in active transcription units. To furtherunderstand the interactions between the PWWP domain of LEDGF and chromatin, NMR spectroscopy and various biophysical methods were utilized. NMR spectroscopy was performedto solve the structure of the PWWP domain of LEDGF. Comparison of the HSQC-‐15N spectra ofthe domain in the presence and absence of native nucleosomes was done to identify PWWPdomain residues interacting with the native nucleosomes. Multiple biophysical methods wereused to prove the LEDGF PWWP domain is sufficient for nucleosome binding and directlyinteracts with the modified core histone proteins. The PWWP domain contains a hydrophobiccavity similar to other known PWWP domain structures. NMR titration experiments of thePWWP domain and nucleosomes identified a region of the protein potentially interacting withnucleosomes. Residues were mapped to the newly solved LEDGF PWWP domain structure and abinding model was proposed involving both protein and DNA interactions. Currently, proteomic-‐based experiments are being performed to identify the histone core post-‐translationalmodification recognized by the LEDGF PWWP domain. Experimental findings have the potentialto aid in the development of novel antiretroviral therapies to prevent the spread of HIVinfection.


THE ROLE OF MICRORNA-‐205 IN BREAST CANCER

Ola Elgamal, Jong-‐Kook Park and Thomas D. Schmittgen

Division of Pharmaceutics, College of Pharmacy, The Ohio State University, Columbus,Ohio.


Purpose: Breast cancer is one of the leading causes of death in women of all races.Triple negative breast cancer (TNBC) is considered the most challenging type of breastcancer as it has the worse prognosis and can not be targeted with the most currentlyavailable treatments. It is crucial to find the molecular changes that cause TNBC. Theintent of this study is to investigate the role of microRNA-‐205 (miR-‐205) in TNBC.

Methods:miR-‐205 was measured in five breast cancer cell lines of four differentclassifications. Classification is based on the expression of estrogen receptor (ER),progesterone receptor (PR) and her2/neu receptor. miR-‐205 was absent in the TNBC celllines MDA-‐MB-‐231 and BT549. Using the miRNA target program TargetScan, gene X waspredicted to have two binding sites for miR-‐205. miR-‐205 – gene X interaction wasvalidated by luciferase reporter assay. 2860 base pairs of the gene X 3’untranslated region (3’ UTR) was cloned downstream of the luciferase gene. MDA-‐MB-‐231 cells weretransfected with gene X 3’UTR luciferase construct and precursor miR-‐205 oligonucleotide (pre-‐miR-‐205 oligo) or control oligo.

Results: In this pilot study, miR-‐205 reduced the expression of the luciferase gene X by30-‐fold compared to control oligo.

Conclusion: miR-‐205 is one of the microRNAs regulating gene X which is reported tohave a role in DNA replication and metastasis. Over-‐expression of miR-‐205 results inreduced luciferase gene translation. Therefore, miR-‐205 has a potential role indecreasing metastasis in breast cancer.


ANTICANCER ACTIVITY AND SAR STUDIES ON VARIOUS SUBSTITUTEDNAPHTHOQUINONES

Jonathan Etter, Deepak Bhasin, Somsundaram Chettiar, May Mok and Pui-‐Kai LiCollege of Pharmacy, Dept of Medicinal chemistry & Pharmacognosy, The Ohio StateUniversity, 500 west 12th, Columbus, Ohio, 43210.


Large numbers of quinones and anthraquinones have been associated with antitumor,antibacterial, antimalarial and antifungal activity. Our initial studies on STAT3 inhibitorsled to the discovery of LLL-‐12 as a potent compound exhibiting antitumor activity.Among the naphthoquinones, LLL-‐3.1 was found to be the most potent two ringanalogue synthesized in that series. In an effort to further analyze the activity anddevelop the SAR, various compounds based on Juglone, Plumbagin, Menadione havebeen synthesized. The synthesized compounds were tested on various cancer cell linessuch as HT-‐29, DU-‐145 and MDA-‐MB-‐231 using MTS assay. The details will be presentedin detail.


ELECTROSPRAY ENCAPSULATION OF SUBUNIT VACCINES IN POLYMERMICROPARTICLES

Tony D. Duong1, Sadhana Sharma2, Kevin Peine3, Gaurav Gupta4, Abhay Satoskar4,Matthew D. Gallovic1, Eric M. Bachelder2, Barbara E. Wyslouzil1, Kristy M. Ainslie1,2

1William G. Lowrie Department of Chemical and Biomolecular Engineering, College ofEngineering, The Ohio State University, Columbus, OH 43210. 2Division ofPharmaceutics, College of Pharmacy, The Ohio State University, Columbus, OH 43210.3Molecular, Cellular and Developmental Biology, The Ohio State University, Columbus,OH 43210. 4Department of Pathology, College of Medicine, The Ohio State University,Columbus, OH 43210.


Vaccines are the most effective intervention for the prevention of disease, and subunit(i.e. protein based) vaccines are considered to be the safest. To generate an enhancedT-‐cell response from a subunit vaccine, researchers have used delivery systems likepolymer microparticles in conjunction with an adjuvant. Our laboratory synthesizesmicroparticles using acetalated dextran (Ac-‐DEX). Ac-‐DEX is unique from otherbiopolymers because it is acid sensitive, its degradation is tunable, and its by-‐products are pH neutral. The acid sensitivity of Ac-‐DEX facilitates stability at extracellular pH (7.4)and rapid degradation at acidic conditions. Since the endosomes of antigen presentingcells (APCs) contain toll-‐like receptors (TLR), whose stimulation results in an adjuvantresponse, and exist at an acidic pH, Ac-‐DEX particles are ideal for delivering a TLRagonist to these immune cells. Ac-‐DEX microparticles have previously been fabricatedthrough emulsion methods. However, severe concentration limits for drugencapsulation are reached for the TLR agonist resiquimod because it can easily partitioninto the continuous phase of an emulsion. An alternative method for resiquimodencapsulation is electrohydrodynamic spraying (i.e. electrospraying). Electrospray is amethod commercially used for ionizing biological molecules for mass spectroscopy, andit is a research technology that continues to be more widely applied in the fabrication ofdrug delivery vehicles. The electrospray process creates minimal shear stresses, can beoptimized to produce a monodisperse population of particles, displays highencapsulation efficiency, and can be operated at ambient conditions. Additionally, itholds the potential to be scaled-‐up to a continuous commercial process (as opposed to abatch emulsion). Current efforts in our lab have shown the successful encapsulation ofresiquimod in Ac-‐DEX particles using electrospray and promising in vivo results for thetreatment of leishmaniasis using these particles. Continuing work is being conductedfor encapsulating an antigenic protein via electrospray, and future work will co-‐deliveran adjuvant and antigen, either in a co-‐encapsulation or separately.


THE NOVEL PROTEASOME INHIBITOR CARFILZOMIB FUNCTIONSINDEPENDENTLY OF P53 AND NF-‐ΚB TO INDUCE POTENT CYTOTOXICITY IN PRIMARY CHRONIC LYMPHOCYTIC LEUKEMIA CELLS

Sneha V Gupta1, Ellen Sass2, Erin H Hertlein2, Jason Dubovsky2, Rosa Lapalombella2,Jennifer Woyach2, Amy Lehman3, David Jarjoura3, John C Byrd 1,2 , David M Lucas 1 2

1College of Pharmacy; 2College of Medicine; 3Center for Biostatistics, The Ohio StateUniversity, Columbus, OH USA


Carfilzomib (CFZ) belongs to a new class of irreversible proteasome inhibitors thatspecifically target the chymotrypsin-‐like subunit (CT-‐L) of the 26S proteasome. CFZ iscurrently in a phase I clinical trial at The Ohio State University in patients with relapsedor refractory CLL, however, the mechanism of action is unknown. Here we demonstratethat a short (1 hr) exposure of 100 nM CFZ is sufficient to inhibit the CT-‐L subunit in CLLcells. This exposure is also rapidly cytotoxic, inducing apoptosis in approximately 50% ofcells by 24 hr. Unlike the reversible proteasome inhibitor bortezomib, the cytotoxicity ofCFZ is not diminished in media with human serum compared to fetal bovine serum.Additionally, CFZ is more cytotoxic to normal CD19+ B cells than normal CD3+ T cells atclinically relevant concentrations of 33 to 300 nM. CFZ causes cell death ex vivo by acaspase-‐dependent apoptotic pathway, indicated by PARP cleavage and rescue by thebroad caspase inhibitor Boc-‐D-‐fmk. Importantly, our studies indicate that CFZ causescytotoxicity in primary CLL cells irrespective of p53 status. The NF-‐kB signaling pathwayis broadly implicated in CLL cell survival and proteasome inhibitors have been reportedto block this pathway via inhibition of IkB degradation. Paradoxically, our resultsindicate that CFZ leads to activation of NF-‐kB, as evidenced by increased nuclearaccumulation of the p50 and p65 subunits of NF-‐kB, as well as phosphorylated IkBα. Thiscorrelates with enhanced binding of the p50/p65 heterodimer to an NF-‐kB probe in anelectrophoretic mobility shift assay. However, despite this apparent NF-‐kB activation, notranscriptional increases were observed in typical NF-‐kB targets genes. This is the firststudy suggesting that treatment with a proteasome inhibitor induces a defective NF-‐κB response in CLL cells. Collectively, our data indicate that proteasome inhibition is arelevant therapeutic target in CLL and supports the development of CFZ for thetreatment of CLL.


Pharmacokinetics and Blood-‐Brain Barrier Penetration of Bendamustinein Patients with Brain Metastases from Solid Tumors

Lei He1, John Grecula2 Yonghua Ling1, Xiaoxia Yang1, Michael Enzerra1, Jeffrey Cotrill1, MarioAmmirati2, Kari Kendra2, Robert Cavaliere2 , Nina Mayr2, Ewa Mrozek2, John McGregor2, ThomasOlencki2, Ennio Chiocca2, James Elder2, Mani Matharbootham2, Tim Lautenschlaeger2, ChimoOluigbo2, Barbara McCracken-‐Bussa2, Laura Mayer 2, Lai Wei2,Mitch Phelps11College of Pharmacy, 2James Cancer Hospital/ OSU Comprehensive Cancer CenterThe Ohio State University, Columbus OH 43210


Objectives of the study: Bendamustine (BM) is a dual functioning DNA alkylating and cross-‐linkingagent approved by the United States FDA in 2008 for treatment of chronic lymphocytic leukemia andresistant/refractory non-‐Hodgkins lymphoma. It is currently in clinical evaluation for treatment ofmany other malignancies. Our team is evaluating BM as a radiosensitizer combined with fractionated stereotactic radiotherapy for treatment of patients with 1-‐4 brain metastases from solidmalignancies in a phase 1 study. While this and other trials are evaluating BM therapy for brainmalignancies, the ability of BM to penetrate the blood-‐brain barrier (BBB) has not been directlydemonstrated in humans. The purpose of our study was to evaluate BM pharmacokinetics and determine if BM is reaching the target metastatic brain lesions by quantifying BM in resected tumors.Methodology: Patients received 30 min intravenous infusion of BM days 1 to 3 with surgical resection of brain lesions on day 3 following BM dosing. Plasma samples up to 2 hours post infusionwere collected on day 1 or 2. Surgeries were conducted after infusion on day 3 with collection ofplasma immediately before, during and after surgery. Cerebral-‐spinal fluid (CSF) and brain tumortissues were also collected. Two highly sensitive LC-‐MS/MS assays for quantification of BM in humanplasma and mouse brain tissue (as a surrogate for human brain tissue) with lower limits ofquantification (LLOQ) of 5.6 and 13.9 nM separately, were developed and fully validatedResults and Conclusions: Two highly sensitive LC-‐MS/MS assays for quantification of BM in humanplasma and mouse brain were fully validated and applied in quantification of BM levels in patients’plasma and brain tissues Plasma pharmacokinetic data from 6 patients treated with 40 mg/m2 BMdemonstrates peak concentrations between 10 and 15 µM at the end of the 30 min infusion withrapid decline to low nM levels within 2 hours and an estimated terminal half-‐life of 0.8 hour, all ofwhich are consistent to previous clinical studies. For the first 3 patients, CSF and brain tumor lesionswere collected from 4.5 to 9 hours after BM infusion. Although BM was detected in the earliestsamples, drug levels were below LLOQ and therefore could not be quantified. By reducing theinterval between BM dosing and surgery, both CSF and tumor lesions of the 3 later patients werecollected approximately 2 hours after the BM infusion. BM levels were quantified between 25 and129 nM. These interim results of our study indicate BM can be detected in CSF and tumor lesions,suggesting BM may serve as a potential radiosensitizer in brain metastases treatment combined withfractionated stereotactic radiotherapy. Further studies are required to explore theconcentration/accumulation of BM which is sufficient for sensitization of the tissue to radiotherapy.Acknowledgements: Approved and funded by the National Comprehensive Cancer Network (NCCN)from general research support provided by Teva Pharmaceutical Industries; Supported in part by NCIP30 CA 16058 and the Pharmacoanalytical Shared Resource, Comprehensive Cancer Center, The Ohio State University.


THE NOVEL ROLE OF INTEGRIN-‐LINKED KINASE IN IL-‐6 INDUCEDTUMORIGENESIS

En-‐Chi Hsu1, Yu-‐Chou Tseng1, Yo-‐Ting Tsai1, Su-‐Lin Lee1, Nicholas J. Sullivan2, Tatiana M.Oberyszyn2, Samuel K. Kulp1, and Ching-‐Shih Chen11Division of Medical Chemistry and Pharmacognosy, College of Pharmacy; 2Department of Pathology, The Ohio State University, Columbus, Ohio, USA


The interleukin 6 (IL-‐6) is a cytokine that play important role in the inflammation.Recently, it has been demonstrated that IL-‐6 induced cancer stem cells via activation ofSTAT3 and NF-‐κB pathway. Integrin-‐linked kinase (ILK) is a kinase that links the cell-‐adhesion receptors, integrins and growth factors to the actin cytoskeleton and regulatesmultiple signaling pathways. ILK was highly expressed in various cancers. One interestingfinding in our lab is that ILK expression is tightly associated with Notch1 activation andIL-‐6 amounts in breast cancer cell lines. Cancer stem cells are related to aggressivephenotype of cancer such as potent cancer initiation, drug resistance and metastasis. The objective of our work is to identify the mechanism of regulation between IL-‐6, ILK,and Notch1 activation. Finally, we figure out that ILK plays important role in the IL-‐6-‐NF-‐κB positive signaling loop and downstream Notch1 activation, which maintain the self-‐renewal ability of the cancer stem cell. Moreover, It would be a novel strategy to targetIL-‐6 caused carcinogenesis via ILK inhibitor, T315.


OSCILLATING AQUEOUS GEL SYSTEMS

Tien-‐Lu Huang and Sylvan G. FrankDivision of Pharmaceutics, College of Pharmacy, The Ohio State University, Columbus,OH 43210


Purpose: Highly viscoelastic aqueous liquid crystal gels formed by surfactant/oil/watersystems are being studied to evaluate their potential as novel drug delivery systems.They have a unique oscillating property that is being investigated as a means ofcontrolling drug release due to the resulting structural changes in these systems.Methods: Based on the phase behavior of various surfactant/oil/water systems, gel systems with suitable oscillation properties and homogenous physical appearance wereselected as prototype drug delivery systems. Microscopy images were examined todetermine the structure and texture of each aqueous gel. Sonograms of the frequenciesand amplitudes of oscillation have been recorded with a virtual digital oscilloscope. Inaddition to mechanical means, electrical currents were applied to stimulate the gels.Lidocaine and lidocaine/prilocaine eutectic mixture were the initial model drugcomponents. Studies of the diffusion of lidocaine from these gel systems wereperformed using Franz diffusion cells, and drug concentrations were analyzed by HPLC.To enhance diffusion, a rotating spindle, or ultrasound was used to disrupt the structureof the gels. Drug release profiles due to different perturbation methods were evaluated.Results: The prototype gel systems are ternary systems of nonionic surfactant andcosurfactant with oil and water content in a variety of proportions. Optical and polarizedlight microscopy has indicated liquid crystal structures of the systems. Oscillation andresonation of sound by the gels were independent of the presence of the model drugs.Sonograms of the model gels exhibited Bessel-‐type functions with damping acoustic waves, and the characteristic frequencies were about 6 kHz. The amount of drugreleased was influenced by the viscous nature of the systems, the action of the rotatingspindle, the stirring rate in the receiver compartment of the Franz cell, as well as theduration of ultrasonic stimulation.Conclusions: The unique oscillating property of the aqueous gel systems is due to theirthree-‐dimensional liquid crystal structures and resulting rheological properties. Initialdata demonstrates that drug diffusion from the prototype gels is regulated by thedegree of altered gel structure. As such, drug release could be controlled, and theseaqueous gel systems may be potential novel drug delivery systems such as in devicesapplied to the skin.


SYNTHESIS AND NMR ANALYSIS OF O-‐CLOSO-‐CARBORANE DERIVATIVES

Keisuke Ishita, Ahmed Khalil, Rohit Tiwari, Werner TjarksDivision of Medicinal Chemistry & Pharmacognosy, The Ohio State University


Carboranes are caged-‐boron compounds containing carbon and boron atoms. Thesestructures are chemically and biologically very stable. They have been utilized primarilyin BNCT (Boron Neutron Capture Therapy) of cancer. Boron-‐halogen bonds areenzymatically more stable than carbon-‐halogen bonds, and thus, radiohalogenated carboranes can also be used for tumor imaging and radiotherapy. Indeed, muchattention has been paid in recent years to the synthesis of radiohalogenated carboranederivatives1). Since many types of tumor cells overexpress the folate receptors2), it can be safely assumed that radiohalogenated carboranes conjugated to folic acid areselectively delivered to cancer cells and internalized by receptor-‐mediated endocytosisfor tumor imaging and/or therapy.

Thus, the aim of this research is the synthesis and biological evaluation ofradiohalogenated carboranes conjugated to folic acid. In the initial phase of thisresearch project, we have investigated the intriguing NMR-‐spectroscopic characteristicsof halogenated carboranes. For this purpose, 9-‐Iodo-‐o-‐carborane (1) was reacted withsodium bromide and Herrmann’s catalyst in DMF to afford 9-‐bromo-‐o-‐carborane (2). 2was then deprotonated with n-‐BuLi and reacted with tert-‐butyldimethylsilyl (TBDMS)chloride to produce two isomers (3, 4) shown in Scheme 1.3) 1H-‐ and 11B -‐ NMR spectraof compound 1-‐4 as well as some other halogenated carboranes will be discussed.

DMF

NaBr, Herrmann's catalyst 1) n-BuLi 2) TBDMSCl

THF

3: R1 = TBDMS, R2 = H 4: R1 = H, R2 = TBDMS

1 2

I Br Br

R1

R2

Scheme 1. Synthesis of o-‐closo-‐carborane derivatives

: C-H : B-H : B

1) Boron science, Narayan S. Hosmane, CRC Press, 2012.2) Xia W., Low P. S., J. Med. Chem., 53, 6811, 2010.3) Rohit Tiwari (Dissertation, College of Pharmacy, The Ohio State University, 2010).


Activation of Silenced Tumor Suppressor Genes in Prostate Cancer Cells by a Novel Energy Restriction-‐Mimetic Agent

Hsiang-‐Yu Lin,1,3,4 Yi-‐Chiu Kuo, Yu-‐I Weng,2 I-‐Lu Lai,1 Tim H.-‐M. Huang,2 Shuan-‐Pei Lin,4 Dau-‐MingNiu,3,5,* Ching-‐Shih Chen1,*1Division of Medicinal Chemistry, College of Pharmacy, The Ohio State University, Columbus, OH43210, USA 2Human Cancer Genetics Program, The Ohio State University, Columbus, OH 43210,USA 3Institute of Clinical Medicine, National Yang-‐Ming University, Taipei, Taiwan 4Departmentof Pediatrics, Mackay Memorial Hospital and Mackay Medicine, Nursing and ManagementCollege, Taipei, Taiwan 5 Department of Pediatrics, Taipei Veterans General Hospital, Taipei,Taiwan

Rank of first author: Research StaffProgram of first author: Division of Medicinal Chemistry and Pharmacognosy

Backgound. Targeting tumor metabolism by energy restriction-‐mimetic agents (ERMAs) hasemerged as strategy for cancer therapy/prevention. Evidence suggests mechanistic linkbetween ERMA-‐mediated antitumor effects and epigenetic gene regulation.Methods. Microarray analysis showed that novel thiazolidinedione-‐derived ERMA, CG-‐12, andglucose deprivation could suppress DNA methyltransferase (DNMT)1 expression and reactivate DNA methylation-‐silenced tumor suppressor genes in LNCaP prostate cancer cells. Thus, we investigated the effects of a potent CG-‐12 derivative, CG-‐5, vis-‐à-‐vis 2-‐deoxyglucose, glucosedeprivation and/or 5-‐aza-‐deoxycytidine, o DNMT isoform expression, DNMT1 transcriptionalactivation, and expression of genes frequently hypermethylated in prostate cancer. Promotermethylation in these genes was assessed by pyrosequencing analysis. SiRNA-‐mediatedknockdown and ectopic expression of DNMT1 were used to validate DNMT1 as a target of CG-‐5.Results. CG-‐5 and glucose deprivation upregulated the expression of DNA methylation-‐silencedtumor suppressor genes, including GADD45a, GADD45b, IGFBP3, LAMB3, BASP1, GPX3, andGSTP1, but also downregulated methylated tumor/invasion-‐promoting genes, including CD44,S100A4, and TACSTD2. In contrast, 5-‐aza-‐deoxycytidine induced global reactivation of thesegenes. CG-‐5 mediated these epigenetic effects through transcriptional repression of DNMT1,which was associated with reduced expression of Sp1 and E2F1. SiRNA-‐mediated knockdownand ectopic expression of DNMT1 corroborated DNMT1’s role in CG-‐5-‐mediated modulation ofgene expression. Pyrosequencing revealed differential effects of CG-‐5 versus 5-‐aza-‐deoxycytidine o promoter methylation in the genes examined.Conclusions. These findings reveal a previously uncharacterized epigenetic effect of ERMAs onDNA methylation-‐silenced tumor suppressor genes, which may foster novel strategies forprostate cancer therapy.

1. Ibragimova I, Ibanez de Caceres I, Hoffman AM, Potapova A, Dulaimi E, Al-‐Saleem T, Hudes GR, OchsMF, Cairns P. Global reactivation of epigenetically silenced genes in prostate cancer. Cancer PrevRes (Phila) 2010;3(9):1084-‐1092.

2. Wei S, Kulp SK, Chen CS. Energy restriction as an antitumor target of thiazolidinediones. J Biol Chem2010;285(13):9780-‐9791.


IDENTIFICATION OF CHARAPTERIZATION OF AN NOVEL INTEGRIN-‐LINKEDKINASE INHIBITOR

Su-‐Lin Lee, En-‐chi Hsu, Chih-‐Chien Chou, Hsiao-‐Ching Chuang, Samuel K. Kulp, and Ching-‐ShihChenDivision of Medicinal Chemistry, College of Pharmacy and Comprehensive Cancer Center, TheOhio State University, 323 Parks Hall, 500 West 12th Avenue, Columbus, OH 43210, UnitedStates


Objectives of the study: Integrin-‐linked kinase (ILK) is an intracellular adaptor and kinasethat links the cell adhesion receptors, integrins and growth factors to the actin cytoskeletonand is a component of multiple signaling pathways. Recent studies have demonstrated thatILK is highly expressed in various cancers, and that inhibition of ILK expression and activitymay serve as an attractive strategy for cancer therapy. An important substrate of ILK isprotein kinase B (PKB)/Akt, which ILK phosphorylates at the S473 site, thereby contributingto its activation. The objective of our work is to develop a novel potent small moleculeinhibitor of ILK.Methodology: Previously, we developed AR12 (formerly OSU-‐03012), a novel celecoxib-‐derived inhibitor of phosphoinositide-‐dependent protein kinase-‐1 (PDK1) that downregulates PKB/Akt phosphorylation at T308 (known as the PDK1 site) and inducesapoptosis and growth inhibition in cancer cells by blocking the PI3K/Akt pathway. Using AR-‐12 as lead, we synthesized a series of derivatives to generate a small kinase inhibitorlibrary, which was screened for compounds that could specifically downregulate PKB/Aktphosphorylation at S473, which is known as the PDK2 target site. The MTT assay was used to examine cancer cell viability after drug treatment. A radiometric ILK kinase assay wasused to assess the ILK kinase inhibitory activities of the compounds. Cellular protein levelsand phosphorylation status of ILK downstream targets were determined in drug-‐treatedcells by western blot analysis.Results & Conclusions: The screening of our focused AR12-‐derived kinase inhibitor libraryidentified T315 as an efficient inhibitor of PKB/Akt phosphorylation at S473 without alteringphosphorylation at the T308 site. Moreover, T315 demonstrated strong cytotoxicity againstprostate cancer cell lines (PC-‐3 and LNCaP cells; IC50 2.0 and 1.8 µM, respectively), but not primary nonmalignant prostate epithelial cells. To date, the phosphorylation of PKB/Akt atits PDK2 site has been attributed to at least ten candidate kinases, including PKCα/βII,mTORC2, ILK, and ATM. After examination of several signaling pathways by westernblotting, two downstream signaling effectors of ILK, GSK-‐3β and MLC2, were shown to be downregulated by T315 in a dose-‐dependent manner, while downstream targets of mTORC2, another PDK2 candidate, were unchanged by T315 treatment. Importantly, T315also dose-‐dependently inhibited ILK kinase activity (IC5 ~0.6 µM) as determined byradiometric kinase assay. Together, these findings indicate that T315 is a putative ILKinhibitor.


BIOACTIVITY-‐GUIDED ISOLATION OF ANTIOXIDANT AND QUINONEREDUCTASE-‐INDUCING CONSTITUENTS FROM ARISTOTELIA CHILENSIS(MAQUI BERRY)

Jie Li 1, Hee-‐byung Chai1, Ye Deng1, William J. Keller2, and A. Douglas Kinghorn11 Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The OhioState University, Columbus, OH 43210; 2Nature's Sunshine Products, Inc., 1655 N. MainStreet, Spanish Fork, Utah 84660.


Aristotelia chilensis (Mol.) Stuntz (Elaeocarpaceae), Maqui berry, is a native South American evergreen shrub. This species produces small edible purple or black berries that are eaten freshor used for juice, jams, and wine-‐making.1 The antioxidant effect of A. chilensis extracts havebeen evaluated in several investigations using different in vitro assays.2,3 Previousphytochemical studies on A. chilensis have revealed the most abundant constituents as beingphenolic compounds, including proanthocyanidins, anthocyanins, and flavonoids.3,4 In a search for naturally occurring cancer chemopreventive agents and to gain an increasing understandingof the phytochemical composition of common botanical dietary supplements, the ethyl acetate-‐soluble extract of A. chilensis collected from Chile showed significant hydroxyl radical-‐scavenging activity, and thus was fractionated via bioactivity-‐guided isolation. Fifteen knowncompounds (1-‐15), comprising twelve phenolic compounds, an organic acid, and two furanderivatives, were isolated from the active fractions of the ethyl acetate-‐soluble extract. Amongthese isolates, phloroglucinaldehyde 2-‐O-‐ß-‐D-‐glucopyranoside (12) was isolated as a newnatural product, and eleven compounds (1, 3, and 7-‐15) were isolated from A. chilensis for the first time. The structures of these compounds were identified on the basis of spectroscopicmethods, including NMR (1D and 2D) and ESIMS. All pure isolates obtained in the presentinvestigation were evaluated for their hydroxyl radical-‐scavenging and quinone reductaseinduction activities using standard procedures. Twelve compounds (1-‐12) showed potentantioxidant activity in the hydroxyl radical-‐scavenging assay, with cyanidin 3-‐O-‐ß-‐D-‐glucopyranoside (2, ED50 = 0.13 μM) being of the greatest potency. Five compounds (3, 7, 9, 11,and 14) exhibited relatively high quinone reductase-‐inducing activity, with protocatechuic acid(9, CD = 4.1 μM) being of the greatest potency. The present investigation has indicated A.chilensis as being worthy of further investigation for the discovery of cancer chemopreventiveagents and as possible functional food ingredient of natural origin.

1. Schreckinger, M.E.; Lotton, J.; Lila, M.A.; Gonzalez de Mejia, E. J. Med. Food 2010, 13, 233-‐246.2. Cespedes, C.L.; El-‐Hafidi, M.; Pavon, N.; Alarcon, J. Food Chem. 2008, 107, 820-‐829.3. Cespedes, C.L.; Valdez-‐Morales, M.; Avila, J.G.; El-‐Hafidi, M.; Alarcon, J.; Paredes-‐Lopez, O. FoodChem. 2010, 119, 886-‐895.4. He, K.; Valcic, S.; Timmermann, B.N.; Montenegro, G. Int. J. Pharmacogn. 1997, 35, 215-‐217.

Acknowledgments: We would like to thank Mr. John Fowble (College of Pharmacy, the Ohio StateUniversity) for facilitating the acquisition of NMR data. Mr. Mark Apsega from the Mass Spectrometry &Proteomics Facility at the Ohio State University is acknowledged for MS training.


ANTILEISHMANIAL ACTIVITY OF A BISBENZYLISOQUINOLINE ALKALOIDFROM THE ROOTS OF THALICTRUM ALPINUM

C. Benjamin Naman,1 Gaurav Gupta,2 Claudio M. Lezama-‐Davila,2 Raymond W.Doskotch,1 Abhay R. Satoskar,2 and A. Douglas Kinghorn.11Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy and2Department of Pathology, College of Medicine, The Ohio State University, Columbus,Ohio 43210, USA


Leishmaniasis is a disease state caused by infection with intracellular protozoa of thegenus Leishmania. Infections by up to 20 species of Leishmania, including L.amazonensis, L. donovani, L. infantum, L. major, and L. mexicana place an estimated 350million people at risk worldwide.1,2 The disease is classified into three major categoriesof infection; visceral leishmaniasis (VL), cutaneous leishmaniasis (CL), andmucocutaneous leishmaniasis (MCL).2 While untreated CL may be cured by the immunesystem or transformed into MCL, VL is a potentially fatal form of the disease that iscaused in Asia and Africa by L. donovani and in South America and Southern Europe by

3L. infantum. As part of ongoing efforts to discover natural product compounds withantileishmanial activity as potential new drug candidates, we have recently begunscreening a compound library comprising natural products and precursors in their totalsynthesis against L. donovani promastigotes. One such sample, tb-‐00069, displayedantileishmanial activity with IC50 = 0.26 µg/mL. The identity of tb-‐00069 was confirmedby NMR spectroscopy and mass spectrometry to be N-‐desmethylthalrugosidine, abisbenzylisoquinoline alkaloid isolated earlier from the roots of Thalictrum alpinum andshown separately to exhibit antimalarial effects and to be devoid of activity against L.major.4,5 Some other bisbenzylisoquinoline alkaloids have also been shown previouslyto have antileishmanial or antimicrobial activity.6-‐8 This work also represents the firstfull spectroscopic characterization of N-‐desmethylthalrugosidine, since its originalcharacterization was completed by utilization of chemical degradation techniques.4

(1) den Boer, M.; Argaw, D.; Jannin, J.; Alvar, J. Clin. Microbiol. Infect. 2011, 17, 1471-‐1477.(2) WHO. The disease and its Epidemiology.http://www.who.int/leishmaniasis/disease_epidemiology/en/index.html. (Accessed March 22, 2012)(3) Croft, S. L.; Olliaro, P. Clin. Microbiol. Infect. 2011, 17, 1478-‐1483.(4) Wu, W.-‐N.; Beal, J. L.; Doskotch, R. W. J. Nat. Prod. 1980, 270, 372-‐381.(5) Ropivia, J.; Derbré, S.; Rouger, C.; Pagniez, F.; Le Pape, P.; Richomme, P.Molecules 2010, 15, 6476-‐6484.(6) Fournet, A.; Barrios, A. A.; Muñoz, V.; Hocquemiller, R.; Cavé, A. Phytother. Res. 1993, 7, 281-‐284.(7) Chan-‐Bacab, M. J.; Peña-‐Rodríguez, L. M. Nat. Prod. Rep. 2001, 18, 674-‐688.(8) Schiff, P. L. J. Nat. Prod. 1981, 46, 1-‐43.


http://www.who.int/leishmaniasis/disease_epidemiology/en/

We would like to acknowledge Mr. Tony Gromovsky for his assistance on this project.

NF-‐ΚB-‐MEDIATED ZIP8 EXPRESSION CONTRIBUTES TO CD-‐INDUCED CELLTOXICITY AND EMPHYSEMA IN MICE

Jessica R. Napolitano1,2 , Mingjie Liu2, Shengying Bao2, Michael Borchers3, Daniel Nebert3,Patrick Nana-‐Sinkam2, Melissa Crawford2, Daren Knoell21Integrated Biomedical Science Graduate Program, 2Dorothy M. Davis Heart and LungResearch Institute, The Ohio State University, Columbus, OH 432103Department of Environmental Health, University of Cincinnati, Cincinnati, OH 45221

Rank of first author: Graduate StudentProgram of first author: Integrated Biomedical Science Program (IBGP)

The zinc transporter SLC39A8, a.k.a. ZIP8, is a primary transporter of cadmium (Cd) entryinto cells. Cd is a heavy metal and carcinogen that is abundant in cigarette smoke andcontributes to smoking-‐related lung disease. The mechanism(s) by which Cd causes lungrelated pathology remain unclear. Our group discovered that ZIP8 is under thetranscriptional control of the NF-‐κB pathway and we have shown that the vitalmicroutrient zinc competes with Cd for entry into lung epithelia through ZIP8. In thisinvestigation we further determined whether ZIP8 contributes to Cd-‐mediated pathogenesis in vitro and in vivo using a mouse model of chronic cigarette smokeexposure. The human A549 lung epithelial cell line and primary human upper airwayepithelial cells (HUAECs) were stimulated with TNFα overnight and then exposed to Cdfor 24-‐48 hours followed by a variety of analyses. Human lung tissue samples from adultsmokers and corresponding nonsmokers were evaluated by qPCR and IHC to determineZIP8 mRNA and protein levels. In animal studies C57Bl/6 (control) and BTZIP8-‐3 (ZIP8overexpressing) mice were subject to chronic cigarette smoke exposure for 4 monthsand then the left lung was perfused, fixed and scored for emphysema. The right lungwas analyzed for mRNA, protein and metal concentrations. In cell culture we observedthat Cd-‐mediated cell toxicity is dependent on ZIP8 expression and that zinc prohibitscell damage. Interestingly, ZIP8 primarily localized to the apical membrane of fullydifferentiated, polarized HUAECs. Consistent with these observations, there was asignificant increase in ZIP8 mRNA and protein levels observed in the lung of smokerscompared to non-‐smokers. Most striking, BTZIP8-‐3 mice in comparison to controlC57Bl/6 adult mice developed more severe emphysema following chronic cigarettesmoke exposure despite similar lung tissue Cd concentrations. This corresponded withan increase in MMP mRNA expression in BTZIP8-‐3 mice. From these studies weconclude that ZIP8 expression influences lung cell toxicity and disease progression invivo. Taken together, we contend that cigarette smoke, which contains high levels ofcadmium, favors a microenvironment in the airway that may promote inflammation,cadmium uptake, and tissue damage.

This work was supported by RO1 HLO86981-‐01 (DLK).


ENDOGLIN REGULATES P38 MAPK-‐INDUCED STRESS FIBER FORMATIONAND ENDOTHELIAL MIGRATION

Chris Pan, Jeff Bloodworth, Mythreye Karthikeyan, Arun Sharma, and Nam LeeChris Pan (Ph.D. student in Pharmacology)

Rank of first author: Graduate StudentProgram of first author: Division of Pharmacology

Endoglin is an endothelial-‐specific TGF-‐β co-‐receptor essential for angiogenesis andvascular remodeling. Endoglin signals through the canonical Smad1/5/8 and Smad2/3pathways to regulate key aspects of angiogenesis, including endothelial cellproliferation, migration and differentiation. While endoglin dysfunction contributes tonumerous vascular conditions, including preeclampsia and hereditary hemorrhagictelangiectasia (HHT), the underlying mechanisms remain poorly defined. Here we reporta novel mechanism by which endoglin controls migration. We demonstrate thatendoglin inhibits cell motility by suppressing p38 MAPK-‐induced actin stress fiberformation. Biochemical and immunofluorescence studies show that endoglin requiresinteraction with its binding partner, b-‐arrestin2, to down-‐regulate p38 MAPK activity,stress fiber formation, and migration. These findings are consistent with our previousdata that endoglin promotes endothelial differentiation and cell survival but antagonizesmigration. Our study identifies p38 MAPK as a novel endoglin signaling target anddefines crucial new role for endoglin in cytoskeletal remodeling.


UNDERSTANDING THE MECHANISM OF ACTION OF ANTI-‐LEISHMANIAL ARYLIMIDAMIDE DB766

Trupti Pandharkar1, Frederick S Buckner2, David Boykin3, Karl A Werbovetz1

1Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio StateUniversity, Columbus, OH, 2University of Washington, Seattle, WA, 3Georgia StateUniversity, Atlanta, GA,


The design of anti-‐leishmanial arylimidamides (AIAs) was originally inspired by diamidineantimicrobial agents such as pentamidine. We recently reported that the AIA DB766(2,5-‐bis[2-‐(2-‐propoxy)-‐4-‐(2-‐pyridylimino)aminophenyl]furan hydrochloride) displayedoutstanding activity against Leishmania donovani intracellular amastigotes [IC50 = 0.036µM].1 DB766 also displayed oral efficacy in murine and hamster models of visceralleishmaniasis [71% and 89% reduction in liver parasitemia when given po at 100mg/kg/day × 5, respectively]. We have performed a series of experiments to betterunderstand the mechanism of action of AIAs. Ultrastructural studies suggest that thetarget of AIAs is distinct compared to diamidines and the cellular effects of AIAsresemble those of sterol biosynthesis inhibitors in Leishmania. To obtain furthermechanistic information, we have developed L. donovani axenic amastigotes that areover 10-‐fold resistant to DB766. Our in vitro susceptibility assays show that thisresistance is not reversed by verapamil, indicating that the overexpression of effluxpumps is unlikely to be responsible for resistance. Further, these DB766 resistantparasites (766R) are twice as sensitive to miltefosine and more than 1000-‐fold moresensitive to ketoconazole and posaconazole than the wild type parasites.Downregulation of CYP51 (sterol 14α-‐demethylase), a well-‐established target of azolesin fungi and Leishmania, has been shown to hypersensitize these cells to the lethaleffects of azoles and miltefosine, respectively.2,3 We thus sought to investigatemodulation of CYP51 and other sterol biosynthetic enzymes in Leishmania as aconsequence of acquired resistance to DB766. Although we did not see any significantchange in the expression levels of CYP51 protein in wild type vs 766R, we observed adramatic decrease in the expression of CYP5122A1, a cytochrome P450 proteinassociated with ergosterol metabolism in Leishmania3, in our DB766 resistant parasites.Studies to investigate the contribution of other sterol biosynthetic enzymes in thedevelopment of resistance to DB766 and mechanism of action of this potent anti-‐leishmanial compound are ongoing in our laboratory.

1. Wang, M. Z., X. Zhu, et al. (2010), Antimicrob Agents Chemother 54(6): 2507-‐16.

2. Mellado, E., Garcia-‐Effron, G., Buitrago, M. J., Alcazar-‐Fuoli, L., Cuenca-‐Estrella, M., and

Rodriguez-‐Tudela, J. L. (2005), Antimicrob Agents Chemother 49(6): 2536-‐38

3. Verma, S., Mehta, A., Shaha, C. (2011) PLoS ONE 6(9): e25273


EFFICIENT DELIVERY OF TLR STIMULATORS TO ANTIGEN PRESENTINGCELLS USING ACETYLATED DEXTRAN MICROPARTICLES

Kevin J. Peine1, Eric M. Bachelder1, Zachary C. Vangundy2, Tracy L. Papenfuss2, Kevin L.Schully3, John T. Pesce3, Andrea M. Keane-‐Myers3, Kristy M. Ainslie1

The Ohio State University Department of Pharmaceutics1, The Ohio State University College of Veterinary Medicine2, Naval Medical Research Center3

Rank of first author: Graduate StudentProgram of first author: Molecular, Cellular and Developmental Biology

Subunit or protein based vaccinations are considered to be a safer alternative to othervaccinations that use heat-‐inactivated or mutated forms of a microbe. Subunit proteinsgenerally have little or no ability to actually stimulate an immune response and thusrequire the addition of an immunostimulatory molecule to enhance the activity of thevaccine. Toll-‐like receptors (TLRs) 3 and 9 are proteins found in the lysosomalcompartment of vaccine mediating immune cells (e.g. dendritic cells). TLRs 3 and 9naturally elicit an immune response to double-‐stranded RNA and unmethylated C/Grepeats, respectively. Poly I:C and CpG are synthetic molecules that mimic double-‐stranded RNA and unmethylated C/G units, respectively . Here, we report theencapsulation of these immunostimulatory molecules in acetylated dextran (Ac-‐DEX) microparticles. These particles were then characterized for TLR ligand loading andencapsulation. The ability of TLR ligand encapsulating Ac-‐DEX particles to increase nitricoxide and cytokine responses from RAW 264.7 macrophages and murine bone marrowderived dendritic cells (BMDCs) was then evaluated. Our results indicate that theseagents could be successfully encapsulated into Ac-‐DEX particles and nitric oxideproduction was at or above the level observed with the free ligand control. Future workwill continue to evaluate cytokine response to these encapsulated ligands.

Acknowledgments: The Defense Advanced Research Projects Agency (DARPA) for funding this work


REGULATION OF THE MACROPHAGE ZINC TRANSPORTER ZIP8 IN RESPONSE TO MYCOBACTERIUM TUBERCULOSIS INFECTION

Charlie J. Pyle Pharm.D.1,2,6 , Larry S. Schlesinger M.D.1,2,3,4 , Daren L. Knoell Pharm.D.1,4,5

1The Dorothy M. Davis Heart and Lung Research Institute, 2The Center for Microbial

Interface Biology, 3Department of Microbial Infection and Immunity, 4Department of

Internal Medicine, 5Department of Pharmacy Practice and Administration,6Translational Science Graduate Program

Rank of first author: Graduate StudentProgram of first author: Translational Science Graduate Program

Mycobacterium tuberculosis (M.tb), the pathogen responsible for tuberculosis, currentlyinfects 1/3 of the world’s population and claims nearly 1.8 million lives each year.M.tbis a highly virulent intracellular bacteria that infects macrophages. Critical to theestablishment of a successful infection byM.tb are phagocytosis by the macrophage andsurvival within distinct phagosomes. The capacity forM.tb to enter and persevere in thephagosomal microenvironment depends in part on its exposure to micronutrients. Zincis an intracellular micronutrient required by all eukaryotic and prokaryotic cells. Humancellular zinc metabolism is regulated by 10, ZnT cytosolic-‐zinc efflux transport proteinsand 14, ZIP cytosolic-‐zinc influx transport proteins. The zinc transporter ZIP8 traffics zincacross the plasma membrane and into the cytoplasm in addition to transporting zinc outof intracellular vesicles such as endosomes and lysosomes. Our group hasdemonstrated that ZIP8 is up-‐regulated in human macrophages in response toinflammation. We hypothesize thatM.tb infection of human macrophages will induceZIP8 production thereby reducing zinc availability within phagosomes. Here wedetermined whether ZIP8 expression is induced in human monocytes duringdifferentiation into monocyte-‐derived macrophages (MDMs) and also in MDMsfollowing infection withM.tb orMycobacterium bovis BCG (BCG). Monocytes weredifferentiated into MDMs by cultivation for five days and whole cell lysates werecollected daily. MDMs were exposed to virulentM.tb H37Rv or BCG and incubated for24, 48 or 72 hours before lysis. ZIP8 mRNA and protein levels were determined by Real-‐Time RT-‐PCR and Western blotting. During differentiation, ZIP8 mRNA levels peaked onday two while protein expression increased until day four. MDMs infected withM.tbH37Rv or BCG exhibited a significant elevation of ZIP8 compared to resting cultures.These studies demonstrate that human macrophage ZIP8 expression is induced byM.tbinfection and support future studies that will examine the role of ZIP8 in host defense.

This work was supported by R01 HL086981-‐01 (DLK) & T32 HL7946-‐11 (CJP, MDW)


WATER-‐IN WATER (W/W) EMULSION SYSTEMS

Anita Sharma, Sylvan G. FrankDivision of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy, The OhioState University, Columbus OH 43210

Rank of first author: Graduat StudentProgram o first author: Division of Pharmaceutics an Pharmaceutical Sciences

Water-‐in water (W/W) emulsions are formed by dispersing one aqueous coacervate phase intoanother immiscible aqueous coacervate phase. Such emulsions are inherently kinetically unstable despite their very low interfacial tension and small difference in density between the two phases.These unique and atypical emulsions consisting of two coacervate phases in equilibrium formed bydextran, polyethylene glycol and water, were stabilized by enhancing the viscosity of both thedispersed phase and the dispersion medium with xanthan gum and gellan gum, respectively. Given the oil-‐free, biocompatible aqueous nature of these emulsions, they may be ideal for novel drugdelivery opportunities where oil and surfactant would be contraindicated.

In vitro drug release studies were performed on two model emulsion systems, i.e., one preparedwith a syringe pump specifically designed for emulsification (Model Emulsion 1), and the otherFrench-‐pressed after being syringe pumped (Model Emulsion 2). Lidocaine (0.1 % w/w) was themodel drug. Three different types of release models, i.e., a Franz diffusion cell, dialysis sac, and aUSP Dissolution Apparatus 4 dialysis insert, were used. Drug release profiles from the emulsionsystems using two different sizes of diffusion cells (6.0 and 15.0 mL), and two different types ofmembranes, Millipore® and Nuclepore® of different pore sizes were also evaluated. The drugconcentrations were analyzed by high-‐performance liquid chromatography (HPLC).

The average droplet size obtained with the pump was ~2 µm. French-‐pressing the pump-‐prepared emulsion decreased the average droplet size to ~11-‐13 µm. However, the French-‐pressed emulsion contained some multiple emulsion droplets. The percent drug release increased with increasing pore sizes of the membranes. Millipore membranes of pore size 0.45 µm gaveoptimal release. Only the dialysis sac model discriminated between the two model emulsion systems throughout the study. However, no significant difference was observed in drug releasebetween Model Emulsions 1 and 2 in any of the release models.

Initial data demonstrates that w/w emulsions have the potential to be used as vehicles for drugdelivery. Model Emulsion 1, being simpler both in terms of preparation and the nature of thedroplets, was selected for future studies. The dialysis sac model simulates the potential application of these systems in clinic and therefore, will be explored further. For example, atherapeutic system in which a microdialysis probe and/or fiber is inserted into a tissue of interest can be envisioned for the use of these emulsion systems in a clinical setting.


ALKALOIDS FROM GREWIA PANICULATA WITH CYTOTOXIC AND NON-‐COMPETITIVE NICOTINIC RECEPTOR ANTAGONISTIC ACTIVITIESPatrick C. Still†, Tatiana F. González-‐Cestari‡, Li Pan†, Hee-‐Byung Chai†, James R. Fuchs†,Tran Ngoc Ninh§, Djaja Djendoel Soejarto┴ , Bitna Yi‡, Brandon J. Henderson‡, Popat N. Patil‡, Dennis B. McKay‡, and A. Douglas Kinghorn*,†

†Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio State University,Columbus, Ohio 43210, United States‡Division of Pharmacology, College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, United States§Institute of Ecology and Biological Resources, Vietnamese Academy of Science and Technology, HoangQuoc Viet, Cau Giay, Hanoi, Vietnam┴Program for Collaborative Research in the Pharmaceutical Sciences and Deparment of MedicinalChemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois60612, United States


Grewia paniculata is a large shrub or small tree that grows in several countries in Southand Southeast Asia. In the present study, three new piperidine alkaloids,microgrewiapines A-‐C (1-‐3), the known piperidine alkaloid microcosamine A, 4 and twoknown compounds, liriodenine 5 and 7′-‐(3′,4′-‐dihydroxyphenyl)-‐N-‐[4-‐methoxyphenyl)ethyl]propenamide 6, were isolated from cytotoxic fractions of theseparate chloroform-‐soluble extracts of the stem bark, roots and leaves of G.paniculata. Microgrewiapines A-‐C (1-‐3) showed IC50 values of 3-‐14 µM against the HT-‐29 human colon cancer cell line. These compounds were also evaluated for their potentialnAChR-‐related effects, and showed IC50 values in the low micromolar range in HEKtsA201 cells expressing human α4β2 or α3β4 nicotinic acetylcholine receptors. As aresult of these studies, microgrewiapine A (1) was found to be a moderately selectivecytotoxic agent for cancerous colon cells over normal colon cells, with noncompetitive nicotinic receptor antagonistic activity at nAChR receptors. This study represents thefirst report of these types of biological activity for the constituents of a plant in thegenus Grewia.

This study was supported, in part, by grant P01 CA125066 (awarded to A.D.K.) from NCI, NIH. Professor Jon Lindstrom, University of Pennsylvania, Philadelphia, PA provided HEK tsA201 cells stably expressingeither Hα4β2 or Hα3β4 nAChRs.


CAVEOLIN-‐3 REGULATES SUBCELLULAR TARGETING AND FUNCTION OFKIR2.1 CHANNELS

Zhaogang Yang, Chen Kang, Keli Hu

Division of Pharmacology, College of Pharmacy, The Ohio State Univesity, USA.

Rank of first author: Graduate StudentProgram of first author: Division of Pharmacology

Inwardly rectifying potassium channels, Kir2 channels in particular, are crucial in shapingthe action potential and stabilizing the resting potential. However, little is known aboutthe subcellular localization of the Kir2.1 channels, an important component of IK1 incardiac cells. The present study was designed to determine whether Kir2.1 channels arelocalized to the caveolar plasma membrane and regulated by caveolin-‐3. We found thatabout 40% (43.81±3.75%, n=3) of the cellular content of Kir2.1 was retained in thecaveolin-‐enriched fractions in cardiomyocytes. In contrast, clathrin heavy-‐chain wasdetected across a broad range of the gradient fractions. Measurement of cholesterollevels within each fraction showed cholesterol to be enriched in the cavelin-‐richfractions. Co-‐immunoprecipitation data from cardiac myocytes indicated that Kir2.1 wasassociated with caveolin-‐3. Expression of caveolin-‐3 in HEK293T cells transfected withKir2.1 significantly increased Kir2.1 in the caveolin-‐enriched fraction by 60%(63.26±8.33%, n=3, p<0.01). Further, immunofluorescent staining showed that asignificant portion of Kir2.1 colocalized with caveolin-‐3 in both transfected HEK293T cells and rat cardiomyoctes. In conclusion, we demonstrate that a significant portion ofKir2.1 channels targets to the caveolin-‐enrich plasma membrane of rat cardiacmyocytes, and the caveolae-‐targeting of Kir2.1 channel is regulated by caveolin-‐3.


DISCOVERY OF A NOVEL CLASS OF NEGATIVE ALLOSTERIC MODULATORSOF nAChRs AS A POTENTIAL TREATMENT FOR TOBACCO ADDICTION:STRUCTURE ACTIVITY RELATIONSHIP AND MECHANISMS OF ACTION

Bitna Yi1, Tatiana F. González-‐Cestari1, Brandon J. Henderson1, Martin L. Dalefield1, SihuiLong2, Julian Richard2, Ryan Pavlovicz3, Karl Werbovetz2, Chenglong Li3, R. ThomasBoyd4,and Dennis B. McKay1

1Division of Pharmacology, Ohio Stat University, Columbus, OH, 2Departmen ofChemistry, Ohio State University, Columbus, OH, 3Biophysic Graduate Program, OhioStat University, Columbus, OH, 4Neuroscience, Ohio State University, Columbus, OH

Rank of first author Graduat StudentProgram o first author Division o Pharmacology

Neuronal nicotinic receptors (nAChRs) are pentameric ligand-‐gated ion channels that have beenfocus of intensive drug discovery efforts. With twelve genes encoding each subunit (α2-‐α10

and β2-‐β4), nAChRs exist in multiple subtypes. Improvement in understanding of the linkbetween specific nACh subtypes and various diseases/disorders sets a new stage for drugdiscovery where modulation of nAChRs in a subtype-‐specific manner renders remarkabletherapeutic advantages. Our laboratory has been focusing on the development of antagonists of nAChRs as therapeutic agents to aid smoking cessation. In particular, subtype-‐selectivity towardα4β2* nAChR (*denotes possible additional subunits) is being pursued based on the implicationof the subtype in nicotine dependence. Our approach to develop subtype selective antagonistsof nAChRs is to target allosteric binding sites. Compared to the highly conserved orthostericbinding site, allosteric binding sites exhibit structural diversity among different subtypes, and thereby representing attractive molecular targets to improve subtype-‐selectivity. Previously, ourlaboratory has identified novel class of negative allosteric modulators (NAMs) of nAChRs.Furthermore, pharmacophore model has been generated based o the four NAMs selectivelytargeting the α4β2 nAChR subtype (KAB-‐18, DDR-‐5, DDR-‐13, DDR-‐18). This pharmacophore served as a query for ligand-‐based virtual screening (LBVS) and yielded several promising hits.The study described here focuses on analogs of Hit 1, one of the top ranked hits from this LBVS.As a lead molecule, Hit 1 has potency in the low micromolar range o Hα4β2 nACh with a ~ 5fold preference against Hα3β4 nAChRs. When assessed for its effect on efficacy of agonist epibatidine, decrease in the maximum efficacy was observed, suggesting non-‐competitivemechanisms of action of Hit 1. In order to gain insight into the chemical interactions involved inactivity, structural modifications to the Hit were made and structure-‐activity relationship (SAR)studies were performed. SAR studies suggested that relative selectivity for α4β2 nAChRs isassociated with double-‐bond in the acid portion of the molecules. To obtain information concerning the binding mode of these NAMs, homology modeling and blind docking studies were performed. Blind docking to homology models of the human α4β2 (Hα4β2) nAChRsuggested a potential binding site of these NAMs. In conclusion, a series of SAR and homologymodeling studies have provided insight into relevant structural/chemical features of the NAMsand potential ligand-‐receptor interactions. This information will direct the design of new NAMsof nAChRs with improved potency and selectivity.


APPLICATION OF TRANSLATIONAL PK/PD MODELING TO PREDICTOPTIMAL DOSE OF SIRNA NANOPARTICLES

1,2 1 2,3 1,2Chenguang Zhou , Mitch A. Phelps , L. James Lee , and Robert J. Lee1Division of Pharmaceutics, College of Pharmacy; 2NSF Nanoscale Science andEngineering Center (NSEC) for Affordable Nanoengineering of Polymeric Biomedical Devices (CANPBD); 3Department of Chemical and Biomolecular Engineering, The OhioState University.


The pharmacodynamics (PD) of small interfering RNA (siRNA) significantly depends on thecellular pharmacokinetics (PK) of siRNA, because the site-‐of-‐action for siRNA is located in the cytoplasm. Hence, a key question is whether the in vivo doses will result in sufficient siRNAexposure at the site-‐of-‐action, thereby achieving adequate gene silencing.The present work proposed translational PK/PD model combining cellular PK/PD model, andsystemic PK and biodistribution models (Fig 1). A mechanistic PK model was developed toaccount for the cellular uptake, intracellular trafficking, releasing of siRNA, and siRNAdegradation. The mechanistic PK model was simultaneously fitted to the total cellular and cytoplasmic concentration-‐time profiles. Then simulated area under the cytoplasmicconcentration time curve (AUC) and gene silencing effect was fitted to an Emax model todescribe the PK/PD relationship. The cellular PK/PD model was linked to systemic PK and tumorbiodistribution models to predict in vivo dose-‐response relationships. A sensitivity analysis wasperformed to evaluate the relative effect of model parameters on the in vivo response.The predicted gene silencing response (69.7% inhibition) at mg/kg was consistent withexperimental data (61.1% inhibition) in literatures, suggesting the validity of our model. The model predicted IC50 and IC90 are 1~1.5 and 3.5~4.5 mg/kg, respectively. The sensitivity analysisrevealed that gene silencing response was most sensitive to the rate of cellular uptake, deliverypathway partition, tumor extravasation, and systemic clearance.The translational PK/PD model provided quantitative framework to integrate the knowledge for the biological system and the ADME properties of siRNA nanoparticles. This method can beused to guide formulation design, select optimal dose, and facilitate the translation of preclinical data to human clinical trials.

Fig 1. TranslationalPK/PD model ofnanoparticle-‐mediatedsiRNA disposition.


IMPACT OF A STUDENT PHARMACIST-‐RUN HEALTH SCREENING PROGRAMIN MEDICALLY UNDERSERVED AREAS AS AN ENTRY POINT INTO THEHEALTH CARE SYSTEM

Jennifer L. Rodis, Pharm.D., BCPS, Kristin A. Casper, Pharm.D., Catherine H. Kuhn, Pharm.D., DouglasC. Cornelius, R.Ph., Michelle E. Ross, Pharm.D. Candidate, Kyle A. Munch, Pharm.D. CandidateThe Ohio State University College of Pharmacy and The Kroger Co., Columbus Division

Rank of first author: PharmDProgram of first author: Pharmacy Practice and Administration

Objectives: 1) To provide free student pharmacist-‐run health screenings targeting those patientswith limited or no access to healthcare. 2) To give student pharmacists the opportunity to gainpractical experience in health screenings and have direct interactions with patients at the communitypharmacy. 3) To determine the number of patients referred to a federally qualified health center as adirect result of student-‐pharmacist run health screenings. 4) To follow up with referred patients todetermine the number of patients visiting, or having already visited, a physician.Methods A grocery store pharmacy chain collaborated with a local college of pharmacy toimplement a student pharmacist-‐run health screening program. Four pre-‐determined store locationsparticipating in the screening program are contracted with local, federally qualified health centers(FQHCs) for the 340(b) drug discount program and are located in medically underserved areas. Four-‐hour screenings occurred at these sites every Friday from October 2011-‐May 2012. Students weretrained on policies and procedures and the screening process prior to screening patients. Allpatients completed a waiver and consent form and completed medical and medication historyforms. Under the supervision of a pharmacist, student pharmacists provided free blood pressureand blood glucose screenings, and body mass index assessments for patients 18 years and older.After the screening, each patient was counseled on the results and referred to his or her physician,or a FQHC, for follow-‐up care as appropriate. Student pharmacists followed u with screened patients to assess the impact of their recommendations on follow-‐up with a physician. Descriptivestatistics were used to analyze data.Preliminary Results: A total of 779 patients were screened at 49 screenings events, yielding anaverage of 16 patients screened per event. 723 patients were screened for hypertension, 44 ofwhom had results <140/90 mmHg. Of the 277 patients with a blood pressure >140/90 mmHg, 14had never been previously diagnosed with hypertension. 713 patients received blood glucosescreenings, 620 of whom were found to have non-‐elevated values. Of the 93 patients with anelevated blood glucose reading, 57 had no previous diagnoses of prediabetes or diabetes. total of367 patients received follow-‐up phone call, 226 of whom were successfully contacted. 76 patientsmade an appointment with a physician as a result of the wellness screenings, 25 of whom did notpreviously have a primary care physician.Preliminary Conclusions/Implications: Greater than one-‐third of patients that were referred for careand successfully contacted via follow-‐up phone call accepted the recommendation of the pharmacist and student pharmacist and sought care from primary care physician. Early data indicated that the Kroger wellness screenings helped to serve as an entry point into the health care system for 25patients who screened above goal or self-‐indicated lack of a primary care physician.This study may help to further describe the role of the community pharmacist in bridging thehealthcare gap for medically underserved patients.Special thanks to the following OSU Key Personnel who assisted with study patient follow-‐up:Andrew M. Burns, Allison M. Henry, Daniel Liu, John C. Myers, Andrew R. Richard, Maegan W.Rothermund, Jennifer K. Ward, Michelle E. Zoellner.


It Takes More Than One miRNA to Treat HCCJon C. Henry1, Jong-‐Kook Park2, Tom D. Schmittgen2

1-‐Ohio State University Department of Surgery. 2-‐ Ohio State University College of Pharmacy

Rank of first author: FellowProgram of first author: Division of Pharmaceutics and Pharmaceutical Sciences

Introduction: The involvement of microRNA (miRNA) in solid malignancies has been well detailed over the past decade. miR research is now focusing o their application for the care of oncologypatients. In Hepatocellular Carcinoma (HCC), a unique miRNA signature has been described by usand others in which two miRNA are commonly shown to be disregulated, miR-‐199a-‐3p and miR-‐221.We have demonstrated that miR-‐199a-‐3p by targetting surface protein CD44 decreases theproliferation and invasion of HCC in CD44-‐promenent cell lines. Our lab has also demonstrated that down-‐regulation of miR-‐221 in HCC in vivo prolongs the survival of xenografted mice.It is assumed that single miRNA-‐therapy for a cancer will not be affective on every patient and wouldlikely have improved efficacy in patients and have a broader spectrum of susceptible tumors in combined with additional miRNA therapy or chemotherapy based-‐regimens. Therefore wehypothesized that therapy with miR-‐199a-‐3p, anti-‐miR-‐221, and sorafenib seperately or incombination would have an increased spectrum of susceptible cell lines and would protentially haveincreased efficacy. Methods: Despite the demonstration of anti-‐miR-‐221’s effectiveness in vivo, it’s effect on multiplecell lines in vitro has not been studied. Therefore we first conducted proliferation analysis on 6 HCCcell lines on treatment with anti-‐miR-‐221. From that IC50s of those cell lines susceptable to anti-‐miR-‐221 and those susceptble to miR-‐199a-‐3p were conducted for each compound. IC50 was alsoestablished in all the cell lines for sorafenib. Proliferation was then analyzed with regards tocombination therapy of anti-‐miR-‐221 and sorafenib, miR-‐199a-‐3p and sorafenib, and miR-‐199a-‐3pand antimiR-‐221. The effects were then analyzed for synergy.Results: Anti-‐miR221 treatment on six HCC cell lines showed it to have a decrease of at least 20% in only two of the cell lines: Hep 3B (21% reduction), and Hep G2 (25% reduction) at a set contration of100 nM. All 6 cell lines were susceptible to sorafenib therapy with IC50s as follows: HepG2 11.3 uM, Hep3B 15.3uM, PLC/PRF5 11.3 uM, SNU182 15.3 uM, SNU423 19.4 uM, and SNU 449 uM.Combination therapy with miR-‐199a-‐3p at 50nM and sorafenib therapy at 2 uM was attempted inSNU423 and Snu449 demonstrating an increased effect with a 22% and 97% decreased proliferationcompared to the negative control treated with 20 uM of sorafenib. The cominbation of 50nM ofanti-‐miR-‐221 and 10uM of Sorafenib on HepG2, SNU423, and SNU449 resulted in a 40%, 20%reduction in HepG2 and SNU423 respectfully, but a 10% increase in SNU449 prolifeation comparedto sorafenib and negative control. The combination of miR-‐199a-‐3p and anti-‐miR-‐221 therapy inHepG2 and SNU449 negated the effect of the individuals oligonucleotides on decreasing proliferationof the cells.Conclusions: In HCC, the regulation of miR-‐199a-‐3p or miR-‐221 is unlikely to be beneficially therapyfor all patients. The use of these microRNA in combination with sorafenib does show an a potentialfor increased efficacy for tumors susceptible to the therapy and perhaps an increase spectrum ofsusceptible tumors by using the combination. Further studies pinpointing the IC50 of the miRNAtherapy and the combination therapy will allow the determination of the effect being addative or synergistic.This study shows at a cellular basis the need for personalized medicine in the future to help selecttherpies that will target the specific tumor. It also suggests that miRNA regulation may be used withstandard therapy to perhaps increase the efficacy of therapy in the correctly selected patients.


SYNTHESIS OF CARBORANYL CONJUGATES OF N3-‐SUBSTITUTEDTHYMIDINE AND THEIR EVALUATION AS SUBSTRATES OF RECOMBINANT HUMAN THYMIDINE KINASE 1

Ahmed Khalil,1 Hitesh K. Agarwal,1 Rohit Tiwari,1 Kevin G. Ricks,1 Elena Sjuvarsson,2

Staffan Eriksson, Michael V. Darby,1 Werner Tjarks1

1Division of Medicinal Chemistry & Pharmacognosy, The Ohio State University,Columbus, OH 43210.2Department o Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, 751 23 Uppsala, Sweden.


N3-‐Substituted amino and amido alkyl conjugates of thymidine (dThd) weresynthesized, including several carboranyl alkyl substituted dThd derivatives. The latterwere designed for boron neutron capture therapy (BNCT) of cancer. Amino or amidogroups were incorporated into the spacer between dThd and m-‐carborane to improvethe hydrogen bonding interactions of these derivatives with their target enzyme, humanthymidine kinase 1 (hTK1). The conjugates were evaluated in phosphoryl transfer assayswith hTK1 comparatively with dThd and the 1st generation N3-‐carboranyl dThdanalogue, N5-‐2OH. Among all tested conjugates, compounds 32 (X= CH2, n= 2, m= 3)and 33 (X= CH2, n= 2, m= 4) were the best substrates of hTK1 with phosphorylation ratesof 45% and 37% relative to that of dThd (rPRs). Both compounds were selected for in-‐depth enzyme kinetics studies. The rKcat/Km (Kcat/Km relative to that of dThd) of bothconjugates were 5.4% and 4.7%, respectively. In comparison, the rPR and rKcat/Km valuesof N5-‐2OH in this specific study were 33% and 10.8%, respectively.


EFFECTS OF (5Z)-‐7-‐OXOZEAENOL ON PROSTATE AND BREAST CANCERCELLS

Ulyana Muñoz Acuñaa, Jennifer Wittwera, Sloan Ayersb, Cedric J. Pearce c, Nicholas H.Oberliesc, and Esperanza J. Carcache de Blancoa.aDivision of Pharmacy Practice and Administration and Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio State University, 141N Parks Hall

12th500 W. Avenue, Columbus, OH 43210.b Department of Chemistry and Biochemistry, The University of North Carolina atGreensboro, P.O. Box 26170, Greensboro, NC 27402.cMycosynthetix, Inc 505 Meadowland Drive, Suite 103 Hillsborough, NC 27278.

Rank of first author: Research StaffProgram of first author: Pharmacy Practice and Administration; Division of MedicinalChemistry and Pharmacognosy

As a part of an ongoing investigation of potential anticancer agents from natural origin,the biological and cellular effects on cancer cells of (5Z)-‐7-‐oxozeaenol were studied.(5Z)-‐7-‐oxozeaenol was isolated from a filamentous fungus as the major component withcytotoxic and NF-‐κB activity. The effects of (5Z)-‐7-‐oxozeaenol on the levels of proteinexpression of NF-‐κB, IKK-‐α and IKK-‐β were analyzed by Western blot analysis atincreasing concentrations. The cells treated with (5Z)-‐7-‐oxozeaenol showed a down-‐regulated NF-‐κB response in a dose-‐dependent manner. The NF-‐κB expression for p50 and p65 units was lower in treated cells when compared with the positive control,rocaglamide (50 µg/mL). In addition, IKK-‐β, an up-‐stream mediator in the pathway wasfound to be down-‐regulated in treated cells in a dose-‐dependent manner. Caspase-‐3 was also expressed in a dose-‐dependent manner in treated HeLa cells. Cell cycle analysis was performed on prostate and breast cancer cell lines, DU-‐145, PC-‐3, and MDA-‐MB-‐231. Inhibited proliferation of treated cells was associated with G1-‐phase arrest. Inaddition, (5Z)-‐7-‐oxozeaenol significantly enhanced apoptosis of treated cells throughactivation of caspase-‐3. Our findings also suggest that the compound, (5Z)-‐7-‐oxozeaenol, is a potent inhibitor of the NF-‐κB pathway.


ISOLATION, STRUCTURE ELUCIDATION, AND BIOLOGICAL EVALUATION OF16,23-‐EPOXYCUCURBITACIN CONSTITUENTS FROM ELEAOCARPUS CHINENSIS

Li Pan,1 Yeonjoong Yong, Ye Deng,1 Daniel D. Lantvit,3 Tran Ngoc Ninh,4 Heebyung Chai,1

Esperanza J. Carcache de Blanco,2 Djaja D. Soejarto,3,5 Steven M. Swanson,3 and A. Douglas Kinghorn*,1 1Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The OhioState University, Columbus, Ohio 43210, United States; 2Division of Pharmacy Practiceand Administration, College of Pharmacy, The Ohio State University, Columbus, Ohio43210, United States; 3Program for Collaborative Research in the Pharmaceutical Scienceand Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy,University of Illinois at Chicago, Chicago, Illinois 60612, United States; 4Institute ofEcology and Biological Resources, Vietnamese Academy of Science and Technology,Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam; 5Department of Botany, Field Museum ofNatural History, 1400 S. Lake Shore Drive, Chicago, Illinois 60605, United States

Rank of first author: Post-‐Doctoral ResearcherProgram of first author: Division of Medicinal Chemistry and Pharmacognosy

Eight new 16,23-‐epoxycucurbitacin derivatives, designated as elaeocarpucins A-‐H (1-‐8),and five known cucurbitacins (9-‐13) were isolated from the chloroform-‐solublepartitions of separate methanol extracts of the fruits and stem bark of Elaeocarpus chinensis collected in Vietnam. Isolation work was facilitated using an LC/MSdereplication procedure, and bioassay-‐guided fractionation was monitored using HT-‐29 human cancer cells. The structures of compounds 1-‐8 were determined on the basis ofspectroscopic data interpretation, with the absolute configurations of isomers 1 and 2established by the Mosher ester method. Compounds 1-‐13 were evaluated in vitroagainst the HT-‐29 cell line and using a mitochondrial transmembrane potential assay.Elaeocarpucin C (3), produced by partial synthesis from 16α,23α-‐epoxy-‐3β,20β-‐dihydroxy-‐10αH,23βH-‐cucurbit-‐5,24-‐dien-‐11-‐one (13), was evaluated in an in vivohollow fiber assay, but proved to be inactive at the dose range used.

Support from grant P01 CA125066 from the National Cancer Institute, NIH, Bethesda, MD, isacknowledged.


A PROSPECTIVE, DOUBLE-‐BLINDED, OBSERVATIONAL CLINICAL COHORT STUDY OF THE ASSOCIATION BETWEEN TACROLIMUS EXPOSURE ANDCYP3A4, CYP3A5 GENOTYPES IN ADULT HEMATOPOIETIC STEM CELLTRANSPLANT RECIPIENTS

Ming J. Poi1 Ali Mcbride1, Jigar Trivedi1 Julianna Roddy1, Jiang Wang2 Danxin Wang3 Hong Zhu4,2,9Elizabeth Marek5 Ee Poi5, Wolfgang Sadee3,6 Steve Devine7,8 , Niesha Griffith1,Mitch Phelps

1 Department of Pharmacy, The Arthur G. James Cancer Hospital and Richard J. Solove ResearchInstitute; 2The Pharmacoanalytic Shared Resource, The James Comprehensive Cancer Center;3Department of Pharmacology, College of Medicine, The Ohio State University; 4Division ofBiostatistics, College of Public Health, The Ohio State University; 5Doctor of Pharmacy Program, College of Pharmacy, The Ohio State University; 6Program in Pharmacogenomics; 7Blood and BoneMarrow Transplant Program; Department of Internal Medicine, College of Medicine, The Ohio StateUniversity; 9Department of Pharmaceutics, College of Pharmacy, The Ohio State University .

Rank of first author: FellowProgram of first author: Translational Science

Background. Tacrolimus is a commonly used immunosuppressant in allogeneic hematopoietic stemcell transplantations (HSCT) for rejection prevention. Unfortunately, tacrolimus has a narrowtherapeutic index that may potentiate a wide variety of adverse effects if not properly monitored.Numerous studies have demonstrated substantial inter-‐subject variability in tacrolimus pharmacokinetics. Several factors have been correlated with tacrolimus pharmacokinetic variability,including polymorphisms in cytochrome P450 (CYP) isoenzymes 3A4 and 3A5 genes, the 2 keyenzymes responsible for biotransformation of tacrolimus have received much attention. The *3allele of the CYP3A5 gene, which codes for a polymorphism that results in a splicing defect and the absence of CYP3A5 protein activity was previously associated with higher initial mean tacrolimus Cmin

values and lower dose requirement in various solid organ transplant recipients. Another recent study has associated rs35599367C>T (CYP3A4*22) and increased risk of supra-‐therapeutic tacrolimusplasma concentration early after kidney transplant. To our knowledge, the influence of CYP3A4 andCYP3A5 genotypes on tacrolimus pharmacokinetics and pharmacodynamics has not been thoroughlystudied in allogeneic HSCT population.Objectives. The primary objectives of this study are to determine the effects of CYP3A4*22 adCYP3A5*1 genotypes on the pharmacokinetics of tacrolimus in HSCT patients and develop acomprehensive model for optimal tacrolimus starting dose in patients receiving allogeneic HSCT.Secondary objectives include to investigate the impact of CYP3A4/3A5 interacting medications onthe tacrolimus pharmacokinetics, compare the incidence of acute graft-‐versus-‐host disease (aGVHD)and nephrotoxicity between the genotypes up to 100 days post-‐transplant; and explore the effect of other CYP3A4/3A5 polymorphisms and selected candidate genes on tacrolimus pharmacokinetics inHSCT patients.Study Design. A prospective, double-‐blinded observational cohort study in adult allogeneichematopoietic stem cell transplant recipients at The James Comprehensive Cancer Center.Anticipated accrual period of 2 years, and total accrual of 180 subjects. An assay using liquidchromatography-‐mass spectrometry (LC/MS) is being developed to detect CYP3A4/5-‐mediatedmetabolites in order to determine the impact of CYP3A4/5 genetic variations. The study has beenapproved by the Institutional Review Board (IRB) and subject enrollment will begin in May 2012.

Acknowledgment: Support from NIGMS U01 GM092655 (PI: Wolfgang Sadee)


PHARMACOKINETICS OF FLAVOPIRIDOL IN LYMPHOMAS

Ming J. Poi1 Jeffrey Jones2 Amy S. Ruppert3 Jeffrey Cotrill4 Jia Ji5 Diana Chung6 Niesha Griffith1,

Mitch Phelps 5

1 Department of Pharmacy, The Arthur G. James Cancer Hospital and Richard J. Solove ResearchInstitute; 2Department of Internal Medicine, College of Medicine, The Ohio State University;3The James Comprehensive Cancer Center; 4 The Pharmacoanalytic Shared Resource, The JamesComprehensive Cancer Center; 5Department of Pharmaceutics, College of Pharmacy, The OhioState University; 6Doctor of Pharmacy Program, College of Pharmacy, The Ohio State University.

Rank of first author: FellowProgram of first author: Translational Science

Background. Flavopiridol is a broad cyclin-‐dependent kinase inhibitor that had been shown highly activein patients with refractory chronic lymphocytic leukemia (CLL). Subsequently, a phase I/II was conductedto evaluate the safety and efficacy of flavopiridol in patients with relapsed/refractory non-‐Hodgkin’slymphoma. Methods. Four cohorts of patients (n=46) were included: indolent B-‐cell, mantle cell, intermediate gradeB-‐cell, and T-‐/NK-‐cell. This study was a nonrandomized, dose-‐escalation trial. Flavopiridol was given as a30-‐minute loading dose followed by a 4-‐hour continuous infusion for 4 consecutive weeks every 6 week at3 dose levels. Blood samples were collected for pharmacokinetic (PK) analysis on day 1 and 22 of cycle 1at pre-‐dose, 0.5, 1, 3, 4.5, 6, 8 and 24-‐hr after starting bolus infusion. Flavopiridol quantification wasdetermined using a validated liquid chromatography-‐tandem mass spectrometry assay. Plasmaflavopiridol concentration-‐time data were analyzed using standard non-‐compartmental methods inWinNonlin (Pharsight). Paired t-‐tests were used to evaluate differences in AUC0-‐∞ and Cmax between days 1and 22 of cycle 1. Associations between PK parameters and toxicities or response among assessable patients were tested using one-‐way analysis of variance (ANOVA) and 2-‐sample t-‐tests, or thenonparametric Kruskal-‐Wallis test.Results. Statistical analyses of PK parameters were performed on all enrolled patients with evaluable PKdata (n=40); data from 5 patients were omitted due to ≥ 25% AUC0-‐∞ extrapolation and one patient didnot have PK data collected. A total of 71 PK profiles comprising 484 plasma flavopiridol concentrationswere determined. Among those with PK data on both days 1 and 22 (n=24), the mean AUC0-‐∞ was 10.404± 5.923 and 11.949 ± 5.109 hr*µM, respectively, and the mean Cmax was 1.954 ± 0.886 and 2.071 ± 0.820µM, respectively. Differences in AUC0-‐∞ and Cmax for day 22 versus day 1 were not significant (P=0.38 andP=0.75). There was no significant difference in AUC0-‐∞ among the 3 dose levels (P=0.36). However, theaverage Cmax of dose level 3 (2.436 ± 1.149 µM) was higher than the Cmax of dose levels 1 and 2 combined(1.714 ± 0.624 µM) (P=0.09, equal variances not assumed). There were no large differences in total bodyclearance, apparent volume of distribution during terminal phase and terminal phase elimination half-‐lifeof flavopiridol among the 3 dose levels or among the four disease cohorts. Overall, pharmacokineticsappeared similar to those previously reported for flavopiridol in other hematologic malignancies. Among the 35 patients assessable for toxicity and response, 2 patients experienced tumor lysis syndrome, withno obvious differences in AUC0-‐∞ and Cmax values. Similar to previous finding, the median AUC0-‐∞ in thisstudy was lower in those who did not experience cytokine release syndrome versus those who did (8.98vs. 12.94hr*µM, P=0.10), so was the average Cmax (1.79 ± 0.75 vs. 2.62 ± 1.15 µM, P=0.04). Other than ahigher median AUC0-‐∞ in those who experienced fatigue (10.48 vs. 8.25 hr*µM, P=0.06), no othernoteworthy associations were observed between PK parameters and other selected toxicities (diarrhea,neutropenia, leucopenia). Higher AUC0-‐∞ and Cmax were observed in patients who achieved partial response (PR); the median AUC0-‐∞ in those with PR, stable disease (SD), and progressive disease (PD) wasrespectively, 13.79, 8.24, and 8.90 hr*µM (P=0.04), whereas the difference in mean Cmax was insignificant.


SYNTHESIS OF ELLAGIC ACID PERACETATE AND ANTITUMOR EFFICACY WITH ENHANCEMENT OF IMMUNITY

Yulin Ren1, Min Wei2, Patrick C. Still1, Xiaozhuo Chen3,4,5 , Klaus Himmeldirk3, Michael A.Caligiuri2,6,7, A. Douglas Kinghorn1,7, Jianhua Yu 6,7

1Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy,2Department of Molecular Virology, Immunology, and Medical Genetics, The Ohio StateUniversity, Columbus, Ohio 43210, 3Department of Chemistry and Biochemistry, 4EdisonBiotechnology Institute, and 5Department of Biomedical Sciences, Molecular and Cellular Biology Program, Ohio University, Athens, Ohio 45701, 6Division ofHematology/Oncology, College of Medicine and School of Public Health, and7Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210.

Rank of first author: Postdoctoral Research AssociateProgram of first author: Division of Medicinal Chemistry and Pharmacognosy,

Ellagic acid (EA), commonly found in many fruits of the human diet, has been reportedpreviously to suppress tumor incidence and to inhibit selectivelymethylbenzylnitrosamine-‐induced formation of esophageal O6-‐methylguanine in rats.Previous studies have shown that ellagic acid peracetate (EAPA) exhibits more potentbioactivities than EA, but a comparison of the antitumor potency of EAPA with that ofEA has not been reported. A new synthetic method was developed for the totalsynthesis of EAPA with α-‐pentagalloylglucopyranose produced from methyl gallate andhydrolyzed to EA, which was derivatized to EAPA through acetylation. A subcutaneousB16 melanoma tumor model of C57BL/6 immunocompetent mice was used to evaluatethe antitumor efficacy of the two chemicals. The treatment of EA and EAPA was initiateda week before tumor inoculation and continued for an additional two weeks, using adose of 0.5 mg/kg per mouse. After the treatment, tumors were removed, weighed,photographed, and the average tumor size was calculated and compared. Theexpression of CD107a and the production of IFN-‐γ in natural killer cells and the levels ofwhite blood cells and other immune cells were determined, with the weights of bodies,livers, and spleens of normal mice also being evaluated. The results showed thatadministration of EAPA significantly suppressed B16 melanoma growth inimmunocompetent mice without affecting natural killer cell activity and was moreeffective than EA. EAPA increased white blood cell quantity in several organs or tissuesincluding peripheral blood, bone marrow, and liver, and such effects were greater thanthose of EA. Furthermore, neither compound resulted in any body, liver or spleenweight changes of normal mice, indicating that these agents are non-‐toxic to mice. Thisstudy suggests that EAPA may be investigated further as a new immunity-‐stimulatory anticancer drug candidate with potential low toxicity for cancer treatment.

Partial support is from grant P01 CA125066 from the National Cancer Institute, NIH, Bethesda, MD,MetaCor Pharmaceuticals Inc., and the Edison Program of the State of Ohio.


CYTOTOXICITY, NF-‐κB P65 INHIBITION, AND IN VIVO ANTITUMOREFFICACY OF SESQUITERPENE LACTONES FROM PIPTOCOMA RUFESCENS

Yulin Ren1, Ulyana Muñoz Acuña2, Daniel D. Lantvit3, Francisco Jiménez4, Ricardo García4,Melciades Mejía4, Heebyung Chai1 Judith C. Gallucci5, Norman R. Farnsworth3,, Djaja D.Soejarto3,6, Esperanza J. Carcache de Blanco1,2, Steven M. Swanson3, A. Douglas Kinghorn1.Divisions of 1Medicinal Chemistry and Pharmacognosy and 2Pharmacy Practice andAdministration, College of Pharmacy, and 5Department of Chemistry and Biochemistry, The OhioState University, Columbus, Ohio 43210, 3Program for Collaborative Research in thePharmaceutical Sciences and Department of Medicinal Chemistry and Pharmacognosy, Collegeof Pharmacy, University of Illinois at Chicago, Chicago, Illinois 60612, 4Jardín Botánico Nacional“Dr. Rafael Ma. Moscoso”, Santo Domingo, Dominican Republic, and 6Botany Department, FieldMuseum of Natural History, Chicago, Illinois 60605

Rank of first author: Postdoctoral Research AssociateProgram of first author: Division of Medicinal Chemistry and Pharmacognosy

Piptocoma is a small genus of the plant family Asteraceae that occurs in tropical andsub-‐tropical regions of the Western Hemisphere. There are no previous reports on thechemical constituents of any member of this genus. As part of search for new naturalproduct anticancer agents from diverse organisms, a leaf crude methanol extract of P.rufescens, collected in the Dominican Republic, was found to be cytotoxic toward theHT-‐29 human colon cancer cell line. Using column chromatography guided bycytotoxicity to HT-‐29 cells, several new and known sesquiterpene lactones (SQLs) wereisolated from P. rufescens. The structures of the SQLs were established from their IR,UV, NMR, and mass spectra, and the absolute configurations were determined byanalysis of single-‐crystal X-‐ray diffraction, Mosher ester reactions, specific rotationvalues, NOESY NMR data, and CD spectra. All SQLs were screened in terms of theircytotoxicity against HT-‐29 cells, and some were tested in a NF-‐κB p65 inhibition assay.The antitumor potential of three highly cytotoxic SQLs, goyazensolide, 15-‐deoxygoyazensolide, and ereglomerulide was evaluated in an in vivo hollow fiber assay.The results showed that all the SQLs isolated were highly cytotoxic toward HT-‐29 cells, with 15-‐deoxygoyazensolide (IC50, 0.26 μM) being the most potently active compound.Several SQLs exhibited NF-‐κB p65 inhibitory activity. A cytotoxic compound,goyazensolide, showed significant in vivo antitumor potency, when tested at a dose of12.5 mg/kg (i.p.) in mice, but two other SQLs, 15-‐deoxygoyazensolide andereglomerulide, were inactive in this in vivo assay system, when evaluated up to a doseof 25.0 mg/kg (i.p.) in mice. This study describes for the first time the cytotoxicconstituents of Piptocoma rufescens and evaluation of the in vivo antitumor activity ofgoyazensolide, which shows potential for further study towards new anticancer drugdevelopment.

Supported by grants U01 CA52956 and P01 CA125066 from NCI, NIH.


IN VIVO QUANTIFICATION OF ACTIVE DECITABINE-‐TRIPHOSPHATE METABOLITE: A NOVEL PHARMACOANALYTICAL ENDPOINT FOROPTIMIZATION OF HYPOMETHYLATING THERAPY IN ACUTE MYELOID LEUKEMIA

Hongyan Wang1,4, Ping Chen1, Jiang Wang1, Ramasamy Santhanam2, Josephine Aimiuwu1, VijayaSaradhi UV1, Zhongfa Liu1, Sebastian Schwind2,4, John C. Byrd2,4, Michael R. Grever2,4, Miguel A.Villalona-‐Calero2,4 Rebecca Klisovic2,4, Alison Walker2,4, Ramiro Garzon2,4, William Blum2,4,

Chan1,2,4Kenneth K. and Guido Marcucci 1,2,3,4

1Division of Pharmaceutics, College of Pharmacy, 2Division of Hematology, 3Department ofMolecular Virology, Immunology and Medical Genetics, College of Medicine, 4ComprehensiveCancer Center, Ohio State University

Rank of first author: Research ScientistProgram of first author: Division of Pharmaceutics and Pharmaceutical Chemistry

Objective of the study: Decitabine (DAC) is used for treatment of myelodysplastic syndrome andacute myeloid leukemia (AML). Following cellular uptake, DAC is activated to DAC-‐triphosphate(TP) and incorporated into DNA. The DAC-‐DNA binds DNA methyltransferases (DNMTs), therebyleading to hypomethylation and re-‐expression of epigenetically silenced genes and ultimatelyanti-‐leukemia activity. However, direct evidence of in vivo DAC-‐TP occurrence in DAC-‐treatedpatients has been difficult to demonstrate. Thus, we aimed to develop sensitive and specificquantification method for DAC-‐TP analysis.Methodology: liquid chromatography/tandemmass spectrometry (LC/MS) assay for intracellular DAC-‐TP was developed by adapting methodfor dNTP/NTP quantification. C57/BL6 and FDC-‐P1/Kitmut cells-‐engrafted NOD/SCID mice were treated with an i.v. bolus dose of DAC at 6.5mg/kg. Bone marrow (BM) and spleen samples werecollected 4 or 24 hours after drug administration. Mononuclear cells were obtained from BM ofAML patients treated with 20mg/m2/day DAC intravenously over 1 hour for 10 consecutive days.Protein levels of DNMTs were analyzed by immunoblotting. Results and Conclusions: Thedeveloped assay exhibited excellent accuracy and precision. Following DAC treatment, wedetected DAC-‐TP in AML cells (in vitro) and BM and spleen of normal and leukemic mice (invivo) Downregulation of DNMTs was also demonstrated in cell lines and mice bone marrow. Theclinical applicability of this method was further proved by measuring DAC-‐TP level in BM fromfive DAC-‐treated AML patients. The mean DAC-‐TP levels were 1.0 0.5 and 0.7 0.4 pmol/106

cells on day 1 and ~ day 5, respectively. Individual dose variability was also observed.Significance: Our assay is the first to determine the DAC-‐TP in vivo Although more extensivestudies are needed for correlating DAC-‐TP levels with disease response and resistance to DAC,the intracellular level of DAC-‐TP may serve as an early and novel pharmacoanalytical biomarkerfor designing more effective DAC-‐based regimens and monitoring onset of resistance inindividual DAC-‐treated AML patient. [Supported by NCI grants UO1-‐CA76576 (KKC, GM, RBK and WB), NO1-‐CM-‐2011-‐00070 (MAV) and RO1-‐CA102031 (GM and KKC)]


PHARMACOKINETIC AND TISSUE DISTRIBUTION OF SYNTHETIC MIR181AAND ITS IN VITRO BIOTARGETS-‐MODULATING ACTIVITY

Zhiliang Xie1, Hongyan Wang1,2, Ming Chiu1, Sebastian Schwind2,3 , Christopher Hickey2,3 , NatarajanMuthusamy2,3, Guido Marcucci1,2,3, Kenneth K. Chan1,2,3

1College of Pharmacy, 2Comprehensive Cancer Center, 3College of Medicine, Ohio State University


Objectives of the study: Deregulated microRNAs (miRs) expression contributes to tumor developmentand progression by altering expression of tumor suppressor gene and oncogenes in acute myeloidleukemia (AML). Restoring downregulated miR or antagonizing overexpressed miR by synthetic RNAoligonucleotides represents a novel therapeutic approach. Our previous studies have shown thatdownregulation of miR181a resulted in overexpression of components of inflammasome, TLR4 and IL-‐1βand in turn activates NFκB pathway in AML cells. Indeed, lower miR181a expression is associated with prognostically unfavorable disease. Thus, the use of synthetic 2'-‐methoxyphosphorothiolate-‐miR181a (2’-‐MeOPSmiR181a), either alone or in combination with other molecular targeting compounds and/orchemotherapy to restore its physiological levels in AML cells may represent a potentially novel treatmentfor AML. Thus, suitable pharmacoanalytical methods that test level and distribution of 2’-‐MeOPSmiR181ain plasma and other biological matrices are needed for its clinical development.Methodology: The assay is developed based on a two-‐step hybridization technique, with synthetic 2’-‐MeOPSmiR181a binding to a biotin labeled 9-‐mer longer capture probe followed by detection offluorescence generated from detection probe linked anti-‐Dig-‐alkaline phosphatase system. This methodwas validated in human THP-‐1 leukemia cell lysates and mouse plasma. The mRNA and protein level ofthe molecular targets of 2’-‐MeOPSmiR181a were analyzed by QRT-‐PCR and western blot, respectively, 24hours after introducing this synthetic oligonucleotide into THP-‐1 cells by nucleofection. Pharmacokinetics(PK) study of 2’-‐MeOPSmiR181a was conducted in CD2F1 mice following an i.v bolus dose of 7.5 mg/kg.The plasma and major organ tissues (bone marrow, spleen, liver, brain, kidney, lung and heart) werecollected at 0.08, 0.17, 0.25, 0.5, 1, 2, 4, 8, 24 and 48 hours post-‐injection. 2’-‐MeOPSmiR181a levels inmouse plasma and tissues were measured by our developed ELISA and its PK properties were analyzed byusing WinNonlin computer software.Results and conclusion: Excellent linearity was observed in cell lysate and mouse plasma at aconcentration range of 10-‐5000 pM (R2>0.99). In cell lysate, the within-‐day and between-‐day coefficientof variations (CVs, n=6) were <10% and <19.5%, respectively, at a concentration range of 50-‐5000 pM andtheir corresponding accuracy values were 93-‐107.4% and 105.7-‐115.2%, respectively. In mouse plasma,the within-‐day and between-‐day CVs (n=6) were <15% and <20% at a concentration range of 50-‐5000 pMand the accuracy values were 96-‐104.3% and 93-‐101.4%, respectively. Our in vitro study showed that 2’-‐MeOPSmiR181a, at 1 µM, could efficiently downregulate the expression level of its bona-‐fide targets TLR4and IL-‐1β in THP-‐1 cells. We further demonstrated that in mice following i.v. bolus dose of 7.5 mg/kg, 2’-‐MeOPSmiR181a displayed a two-‐compartmental PK with measured C5min of 7.7 µM, AUC (area under thecurve) of 118.1 min*μM, Beta-‐HL (terminal half-‐life) of 17 hours and the CL (total body clearance) 0.008L/min*kg. In addition, its tissue levels were measurable from 5 minutes to at least 24 hours after dosing. Of note, the intracellular level of 2’-‐MeOPSmiR181a in bone marrow mononuclear cells achieved 1.0-‐2.6 pmol/mg protein (~30nM) during the time that we monitored.Significance: A novel sensitive quantification assay was developed and applied for characterization of 2’-‐MeOPSmiR181a PK and tissue distribution in normal mice. Moreover, the in vitro targets-‐downregulatingconcentrations were achieved in mice plasma in vivo. These data indicated that our method is applicablefor PK-‐PD modeling in leukemic mouse treated with 2’-‐MeOPSmiR181a. This will allow the developmentand rapid translation of this novel compound into the clinic for AML.This study is supported by NIH grant R01-‐CA135332 (KKC, NM and GM).


Simultaneous Quantification of 5-‐Hydroxymethyl-‐2’-‐deoxycytidine andGlobal DNA Methylation in DNA using Liquid Chromatogram TandemMass Spectrometry

Zhiliang Xie, Jiang Wang, Kenneth K. Chan and Zhongfa Liu

Division of Pharmaceutics, College of Pharmacy, the Ohio State University


Objectives of the study: 5-‐Hydroxymethyl-‐2’-‐deoxycytidine (5-‐HmdC) has beenidentified as a new novel DNA methylation marker and base modification of DNA methylation. However, most established DNA methylation methods are incapable ofdistinguishing 5-‐methyl-‐2’-‐deoxycytidine (5-‐mdC) from 5-‐HmdC. Herein, we developed aLC-‐MS/MS method for simultaneous quantification of 5-‐HmdC and 5-‐mdC in DNA todifferentiate these two DNA methylation markers. This method was used to measuregenomic DNA methylation level (GDM) and the content of 5-‐HmdC in somatic tissuesand cancer cells.Methodology: A triple-‐quadruple mass spectrometry was used to quantify 2’-‐deoxycytidine (2-‐dC), 5-‐mdC, 5-‐HmdC using 5,6-‐dihydro-‐5-‐azacytidine as the internalstandard by monitoring the following ion transition channels (m/z): 228>112, 242>126,258>142, and 247>115, respectively. The genomic DNA isolated from mouse livertissues, three leukemia cell lines, two breast cancer cell lines and one pancreatic cancercell line was hydrolyzed, and the levels of 2-‐dC, 5-‐mdC, 5-‐HmdC in these DNAs weredetermined using this method.Results and conclusion: The lower limit of quantification for 2-‐dC, 5-‐mdC and 5-‐HmdC inthe hydrolysates was 0.2 ng/mL. The within-‐day and between-‐day parameters for thequality control concentrations between 0.2-‐1000 ng/mL in DNA samples met the US FDAGLP analytical method criteria with CVs < 8% and accuracy in the range of 89% to 107%.In addition to the detection of 1.8 to 10% GDM in these samples, 5-‐HmdC was alsodetectable in mouse liver tissues (≤ 1/3 of 5-‐mdC) and in Kasumi-‐1 and HL-‐60 leukemia cell lines and MCF-‐7, a breast cancer cell line (≤ 1/20 of 5-‐mdC). However, 5-‐HmdC wasundetectable in the pancreatic cancer cell line, leukemia MV4-‐11 cell line, and breastcancer MDA-‐MB-‐231 cell line. Notably, two-‐week daily oral administration of 1.8 g/kgcurcumin in mice significantly decreased 5-‐HmdC levels in liver tissues without affectingthe GDM.

A specific LC-‐MS/MS method was established to differentiate GDM and 5-‐HmdC in DNA samples, which is an important tool for future DNA methylation analysis.

Supported by BioMedical Mass Spectrometry Laboratory (Liu and Chan) and R21 grant (CA135478) fromthe National Cancer Institute (Liu).


8,8-‐DIALKYLDIHYDROBERBERINES WITH POTENT ANTIPROTOZOALACTIVITY

Xiaohua Zhu,1 Molla Endeshaw,1 Shanshan He,1 Trupti Pandharkar,1 Emily Cason,2 Kiran V. Mahasenan,1 Chenglong Li,1 Mark Bahar,1 A. Douglas Kinghorn,1 Mark E. Drew,1,2 andKarl Werbovetz11Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio StateUniversity, Columbus, OH, 2Department of Microbial Infection and Immunity, College ofMedicine, The Ohio State University, Columbus, OH


The plant quaternary alkaloid berberine continues to be investigated for a host ofbiological effects, including anticancer, antidiabetic, and anti-‐infective activity. Werecently identified a novel semisynthetic berberine derivative, 5,6-‐didehydro-‐8,8-‐diethyl-‐13-‐oxodihydroberberine, that displayed potent antileishmanial,antitrypanosomal and antimalarial activity and showed efficacy in a murine visceralleishmaniasis model.1 However, this compound was toxic to mice when given at i.p.doses higher than 1 mg/kg/day. We prepared analogs of 5,6-‐didehydro-‐8,8-‐diethyl-‐13-‐oxodihydroberberine in an attempt to explore the antiparasitic structure-‐activity relationship of the class and to reduce the in vivo toxicity of the lead compound. Theseanalogs were prepared in three or four steps starting from berberine. Semisynthetic8,8-‐dialkyldihydroberberines (8,8-‐DDBs) possess mid-‐ to low-‐nanomolar potency againstPlasmodium falciparum blood-‐stage parasites, Leishmania donovani intracellularamastigotes, and Trypanosoma brucei brucei bloodstream forms. 8,8-‐Dialkylcanadines,obtained by reduction of the corresponding 8,8-‐DDBs, are much less potent againstthese parasites in vitro. 8,8-‐DDBs show efficacy in a mouse model of visceralleishmaniasis and are less toxic than the lead compound. 8,8-‐DDBs may thus be usefulin discovering new antimalarial, antileishmanial, and antitrypanosomal drug candidates.

1. Bahar, M.; Deng, Y.; Zhu, X.; He, S.; Pandharkar, T.; Drew, M. E.; Navarro-‐Vázquez, A.; Anklin, C.;Gil, R. R.; Doskotch, R. W.; Werbovetz, K. A.; Kinghorn, A. D. Bioorg. Med. Chem. Lett. 2011, 21, 2606.

Acknowledgement -‐ This work was funded by a pilot grant from the OSU Public Health Preparedness forInfectious Diseases program.


USE OF AN ELECTRONIC MEDICAL RECORD TO IMPROVE CARE ANDMONITORING OF CHRONIC KIDNEY DISEASE IN A PATIENT-‐CENTEREDMEDICAL HOMEKelli D. Barnes, Neeraj Tayal, Amy Lehman, Stuart J. BeattyThe Ohio State University College of Pharmacy; The Wexner Medical Center at The OhioState University

Rank of first author: ResidentProgram of first author: Pharmacy Practice and Administration

Purpose: The purpose of this study is to use the electronic medical record (EMR) andpharmacist intervention to identify patients with stage 3, 4, or 5 CKD and improve carewithin a patient-‐centered medical home. Objectives of the study are to increase compliance with the National Kidney Foundation guidelines for monitoring and care ofCKD, ensure appropriate dosing of medications based on patient’s calculated creatinineclearance, determine the percentage of pharmacist recommendations accepted by thepatient’s primary care physician (PCP), and track pharmacist time spent completing theintervention.Methods: The EMR will generate a list of adult patients with an estimated glomerularfiltration rate <60 mL/min/1.73mm2. A retrospective chart review of identified patients willbe performed to: 1) confirm presence of CKD in patients with criteria for stage 3, 4 or 5 CKD,2) assess completion of recommended laboratory monitoring and medication therapy forCKD, and 3) assess appropriate dosing of medications. Pharmacist recommendations forcare will be communicated with the patient’s PCP; patients will be contacted if laboratorymeasures or medication changes are recommended.Preliminary Results: 201 (11.3%) patients with CKD were identified through EMR generatedreports with 54.2% of patients not having CKD listed as a medical problem in the EMR priorto pharmacist intervention. Improvement in all recommended laboratory monitoring andmedication therapy occurred as a result of pharmacist intervention. Additionally, 0.86medications per patient were dosed inappropriately or contraindicated based on thepatient’s renal function; 90.2% of medication recommendations made by the pharmacistwere accepted by the patient’s PCP.Conclusions: Opportunities for the improvement in identification and care of CKD in aprimary care setting exist. Pharmacists are well positioned to work with PCPs to improveCKD care, monitoring, and medication dosing. Future completion of this study will allow forintegration and sustainability of outpatient renal dosing services into primary care settings to improve patient outcomes.Maria Joy, Ashley Shumaker, The American Society of Health-‐System Pharmacists Research and EducationFoundation


IMPACT OF BARCODE MEDICATION ADMINISTRATION ON EMERGENCYDEPARTMENT ERRORSJoseph Bonkowski; Joseph Melucci; Beth Prier; Cynthia Carnes; Jay Mirtallo; Robert WeberWexner Medical Center at The Ohio State University, Columbus, OH


The medication use system is error prone with medication administration accounting for34-‐54% of medication errors. Barcode medication administration (BCMA) improves theaccuracy of medication administration in hospital inpatients, but has limited use inemergency departments (ED); this is due to short lengths of ED stay and limited use ofelectronic medical records (EMR). The Ohio State University Medical Centerimplemented an EMR and BCMA in the ED, allowing the opportunity to study the impactof this technology on medication administrations errors.

A single-‐center, pre/post observational study was conducted to compare medicationadministration errors after implementing BCMA. Naïve observers documentedmedication administration 2 months prior to and 4 months post BCMA. A medicationadministration error was defined as any discrepancy between the administeredmedication and the physician’s order. The primary aim of this study, the medicationadministration error rate, was calculated by dividing the number of medicationadministration errors by the number of medication observations. A secondary aim compared medication administration errors to the time of day and therapeutic class.Medications administered by non-‐nursing staff were excluded from observation. Preand post medication administration error rates are compared using a 2 proportion z-‐test; time of day and medication category differences are calculated using linearregression.

996 medication observations were conducted in the baseline period with an error rateof 6.0%. 956 observations are planned in the post BCMA study period.

Conclusions and results will be presented after data collection and evaluation iscomplete.

Jessica Dempsey, PharmD candidateAneeka Quereshi, PharmD candidateErin Reichert, PharmDJessica Traeger, PharmD candidateJason Walsh, BSN, MBA


IMPACT OF AN AUTOMATIC REFILL SYSTEM ONMEDICATION POSSESSIONRATIOS IN THE COMMUNITY PHARMACY SETTING

Jennifer R. Frerick, Tara R. GreenThe Ohio State College of Pharmacy and Kroger Pharmacy


Medication adherence is directly associated with improved clinical outcomes. Therefore,it is crucial to ensure that pharmacists encourage patients to remain adherent throughavailable channels. Enrollment of patients in an automatic refill system aims to improvemedication adherence by making it easier for patients to fill their medications on time.One useful way to measure this impact is through the medication possession ratio(MPR) which is defined as the ratio of the number of days between the last refill and thenext expected refill to the number of days between the last refill and the next actual fill.Retrospective data will be collected for a random sample of patients enrolled in the automatic refill system at a community pharmacy chain. MPRs will be calculated for sixmonths before and after enrollment to assess the impact of the automatic refill system.Specific disease states for comparison include hypertension, dyslipidemia, diabetes,depression, asthma/COPD, and gastroesophageal reflux disease. Demographic data ofpatients including age, gender, insurance coverage, number of chronic medications, andmethod of refill notification will also be collected and analyzed. Data collection will takeplace in February and March of 2012. Results will provide information to identifypotential areas for improvement in counseling and patient care programs that mayenhance adherence and patient outcomes.


EVALUATION OF PHARMACY FACULTY KNOWLEDGE AND PERCEPTIONSOF THE PATIENT-‐CENTERED MEDICAL HOME (PCMH) CONCEPT WITHIN PHARMACY EDUCATION

Anisha B. Grover, Bella H. Mehta, Jennifer L. Rodis, Kristin A. Casper, Randy K. WexlerThe Ohio State University


Objectives: The Patient Protection and Affordable Care Act of March 2010 emphasizesthe need for a reorganized primary care system and supports patient centered medicalhome (PCMH) as a primary care initiative. Future pharmacists have an importantopportunity to advance practice through PCMH practices, and pharmacy education hasa central responsibility in preparing pharmacists to effectively contribute in this setting.This project aims to 1) assess pharmacy faculty knowledge about key PCMH principles,2) evaluate pharmacy faculty perception of inclusion of PCMH information in didacticand/or experiential pharmacy curriculum, and 3) evaluate pharmacy faculty perceptionof where and how this information should be taught. Methods: A roster of currentpharmacy faculty has been obtained from the American Association of Colleges ofPharmacy (AACP) and used to create a database of potential participants. Acustomizable survey program was used to develop and implement an anonymous,online survey. The survey was pilot tested by a group of non-‐AACP faculty members,and refined based on input. Faculty rated their familiarity with key PCMH definitionsand principles. Participants indicated whether or not PCMH concepts are currentlyincluded and should be included in pharmacy education and if so, where in thecurriculum, required or elective, and how much time should be dedicated to this topic.Demographic information was collected. The survey remained open for one month andtwo reminder emails were sent during the midpoint and final week of the datacollection period, which occurred in February and March 2012. Descriptive statistics willbe used to report responses. Results: Reported outcomes will include information relating to the study objectives. Conclusions: By characterizing pharmacy facultyknowledge and perceptions of PCMH in pharmacy education, it is anticipated thatopportunities for faculty and student education can be identified.

Acknowledgements: Kyle Porter (Biostatistician)


DRUG SHORTAGES: MANAGEMENT AND RESPONSE IN HEALTH-‐SYSTEMPHARMACY

Deron T. Lundy, Jennifer L. Rodis, Rodney G. Wirsching, Milap C. Nahata

The Ohio State University College of Pharmacy and Grant Medical Center

Rank of first author: ResidentProgram of first author: MS Health-‐System Pharmacy Administration

Background/Introduction: The importance of managing drug shortages has increasedover the past decade as the number of drug shortages has also increased. As a result,there is an increased burden on health-‐systems, and changes to clinical practice andinventory management have been occurred. The purpose of this study is to examinethe relationship between hospitals’ perceived success at drug shortage managementwith the level of multidisciplinary and executive involvement in drug shortagemanagement, and adherence to American Society of Health-‐System Pharmacists’ (ASHP)shortage management guidelines.Methods: Prior to commencement, this study was determined to be exempt fromreview by the OhioHealth and Ohio State IRBs. Using the Qualtrics™ online survey tool, asurvey was emailed to 555 directors of pharmacy at non-‐federal, acute care sitesidentified in the publicly available ASHP online residency directory. This survey assessedthe three main study variables, and collected demographic information about therespondent’s position and hospital.Preliminary results: The survey is still open to respondents, so final results are pending.A total of 177 respondents completed the survey as of April 17, 2012, yielding aresponse rate of 32%. 61% of respondents agreed with the statement that theirinstitution successfully managed drug shortages, and only 27% strongly agreed. Patientcare threat assessments occurred more often than financial threat assessments. 95% ofrespondents said that they engage in stockpiling of medications, a practice notrecommended by ASHP guidelines. Respondents reported high frequency ofconsultation with medical and pharmacy staff, but lower frequency of consultation withnursing, hospital executives, and risk management.Discussion: Final results are pending close of survey and full statistical analysis. Theseresults will help evaluate the impact of ASHP guideline utilization and multidisciplinary and executive participation on perceived shortage management success.


MEDICATION EVENT HUDDLES: EFFECT OF AN ELECTRONIC DATABASE ONINTERVENTION FOLLOW-‐UP IN A PEDIATRIC HOSPITAL

Jenna Merandi, Pharm.D., Shelly Morvay, Pharm.D., Dorcas Lewe, RN, Barb Stewart,RN, Char Catt, RN, MS, Jay Mirtallo, RPh, MS, Karl Kappeler, RPh, MS, Sheryl Szeinbach,PhD, RPh

Nationwide Children's HospitalThe Ohio State University


Purpose: To determine the impact of an electronic database on the percent ofinterventions completed following medication event huddles.

Methods: An audit was conducted at a free-‐standing academic pediatric hospital usingretrospective data from the medication event huddle database. Intervention follow-‐up from medication event huddles was assessed between the time periods of March 1,2010, through July 1, 2011. Data collection included the original event report summary,names of medications, staff members involved, location of the event, date ofoccurrence, type of intervention, and the time to completion of each interventionfollowing a medication event huddle. Data were entered into Microsoft Excel®spreadsheet to allow for descriptive statistical analysis. An electronic database wascreated to eliminate the use of multiple systems for huddle management, allow fordocumentation of medication event huddles, and generate automatic reminders toindividuals involved in the huddle/intervention follow-‐up. The primary outcomeassessed was the percent change in completion of intervention follow-‐up afterimplementation of an electronic database. Secondary outcomes included categorizationof interventions from the medication event huddles.

Results: The baseline results of this study indicate only 31% of interventions frommedication event huddles are documented as being completed. The percentage ofinterventions completed or in progress, but not documented as such is unknown.Process changes, education, and order improvements are the most frequent categoriesof huddle interventions. Implementation of a user friendly electronic database couldfacilitate documentation and management of interventions and ultimately increasepatient safety. Database build to be complete by March 1, 2012.

Acknowledgements:Phillip Chanthasene, Information Systems Analyst, Nationwide Children's Hospital


PATIENT-‐CENTERED CARE AT A GENERAL INTERNAL MEDICINE PATIENT-‐CENTERED MEDICAL HOME

Shannon D. Peter, PharmD; Kyle Porter, MAS; Stuart J. Beatty, PharmD, BCPSThe Ohio State University College of Pharmacy


Purpose: To 1) Determine patient perceptions of the degree of patient-‐centeredness ofvisits with a pharmacist, internal medicine resident, attending physician, nursepractitioner, social worker, or any combination of the above at a tier 3 General InternalMedicine (GIM) Patient-‐Centered Medical Home (PCMH) and 2) Examine potentialdifferences of patient-‐centeredness perceptions based on healthcare provider(s)providing care during each visit.

Methods: A convenience sample will be used to recruit GIM patients age 18 years andolder. Data will be collected via a one-‐time electronic 21-‐item Consultation CareMeasure (CCM) questionnaire at the end of a GIM visit with one or more of the specifiedhealthcare practitioners. Reported data will include a total CCM patient-‐centeredness score, as well as scores on each of 5 CCM patient-‐centeredness subscales.Demographics, the amount of time subjects spent with healthcare providers, theamount of time spent waiting, and the length of time each subject has been followed atthe PCMH will also be collected. Comparison data analysis will take place to examinecorrelations between the above items and subject perceptions.

Results: With at least 50 responses in the pharmacist, attending physician, and diabetesclinic practitioner groups, the study will have 80% power to detect a 10% difference inCCM scores between provider types. At the time of abstract submission, a total of 151questionnaire responses have been collected.

Conclusions: We postulate that results will be used to guide future initiativesimplemented to improve patient-‐centered care, and will support new team-‐based healthcare models.


PHARMACY RESIDENTS' PURSUIT OF ACADEMIC POSITIONS

Tiffany R. Shin, Colleen A. Clark Dula, Jennifer L. Rodis, Bella H. Mehta, Maria C.PruchnickiThe Ohio State University College of Pharmacy


Purpose: Determine the percentage of pharmacy residents that accept an academicposition at the end of residency, identify factors influencing residents’ decision topursue/not pursue academia, compare perceived characteristics of the ideal positionearly in residency versus characteristics of positions accepted upon completion.Methods: Study includes PGY-‐1 and PGY-‐2 pharmacy residents, and consists of anelectronic pre-‐/post-‐survey with matched responses. Survey invitations weredisseminated via residency directors in October 2011; residents who provided an emailaddress will receive the May 2012 follow-‐up survey. Job preferences, characteristics ofthe ideal job, interest in academia, and experience in teaching and research wereevaluated in pre-‐survey. End-‐of-‐residency survey will focus on job selection, includingapplied and accepted positions, with specific questions regarding the pursuit ofacademic positions and characteristics of positions accepted by residents. Results: Pre-‐survey had 1002 respondents (approximately 39% response rate), 936 pharmacyresidents were included (71.6% PGY-‐1, 26% PGY-‐2, 2.3% combined program). 46.4% ofresidents agreed they were seriously considering a position in academia, 30% wereneutral, and 22.8% disagreed. Formal training in teaching was available to 71.9% ofresidents, while 26.5% had formal training in precepting and 16.4% in research. The topsettings where residents wanted to work upon completion of residency were inpatientclinical (68%), academia (40%), and ambulatory care (31%). More PGY-‐2 residents(59.8%) than PGY-‐1 residents (30.8%) chose academia as a top two career option(p<0.001). Academia was more likely a top choice for ambulatory care (55.2%) andspecialty inpatient residents (63%) than pharmacy practice (27.7%), managed care(19%), and administrative (33.3%) residents (p<0.05 for all comparisons). Topcharacteristics of the ideal job were collaboration with others (63%), variety of dailyactivities (46%), and free time for leisure/family (35%). Conclusions: Post-‐graduate trainees are ideal candidates for faculty recruitment, with many interested in academia.However, many residents likely need additional training for some responsibilities.

Acknowledgements: Kyle Porter, MAS


MEDICATION ERRORS WITH PARENTERAL NUTRITION: IMPACT OFINGREDIENT SHORTAGES

Michael Storey, PharmD*, Robert Weber, PharmD, MS, BCPS, FASHP, Kelly Besco, PharmD,FISMP, Stuart Beatty, PharmD, BCPS, Kumiko Aizawa, MS, Jay Mirtallo, MS, BCNSP, FASHPThe Ohio State University College of Pharmacy


Background: Ingredient shortages have a significant impact on parenteral nutrition (PN) safety. Due to a lack of appropriate alternatives for PN therapy, the utilization ofunfamiliar products or systems has risen and in some instances has led to harmfulmedication errors. Shortages have affected nearly every component of PN in recentyears. The relationship of PN ingredient shortages to harmful medication errors has notbeen formally evaluated. This study characterizes PN medication errors and correlatesthem with recent medication shortages, with a particular interest in preventable eventswith harm (NCC-‐MERP Index E-‐I) that occurred as a result of PN ingredient shortage.Methods: Medication errors involving PN that were reported to the national,anonymous reporting MED-‐MARX database between May 2009 and April 2011 werereviewed. All errors were categorized by ingredient, node, and severity. Thecategorization of the reported events was validated by an expert panel. A timeline of PNingredient shortages was collected, and compared with the PN errors to determine ifevents could have been directly caused by an ingredient shortage. This information wasused to determine the prevalence and change in harmful PN events during periods ofshortage, determining if a statistically significant difference exists in errors duringshortages as compared with a control period (i.e., no shortage).Results: Parenteral nutrition shortages were associated with thirteen errors; most ofthese were associated with intravenous fat emulsions. Nineteen errors were associated with patient harm. Errors were primarily associated with ordering, transcribing, andadministration nodes.References:1. National Coordinating Council for Medication Error Reporting and Prevention. AboutMedication Errors. 20 Feb 2001. Accessed 20 May 2011. Available online:http://www.nccmerp.org/medErrorCatIndex.html2. ISMP. ISMP Medication Safety Alert! 2011;16(8):1-‐2.3. Hicks RW, Becker SC, Cousins DD, eds. (2008). MEDMARX data report. A report on therelationship of drug names and medication errors in response to the Institute ofMedicine’s call for action. Rockville, MD: Center for the Advancement of Patient Safety,US Pharmacopeia.Acknowledgements:The Institute for Safe Medication PracticesThe University of Utah Drug Information ServiceThe American Society of Parenteral and Enteral Nutrition


http://www.nccmerp.org/types-medication-errors

EVALUATION OF THE RATES AND CHARACTERISTICS OF ABANDONEDPRESCRIPTIONS PRESCRIBED BY FEDERALLY QUALIFIED HEALTH CENTER PROVIDERS AT 340B CONTRACTED COMMUNITY PHARMACIES

Shannon L. Yarosz, PharmD, RPh; Catherine H. Kuhn, PharmD, RPhThe Ohio State University College of Pharmacy; Kroger Patient Care Center


Purpose: Federally Qualified Health Centers (FQHCs) are eligible to participate in the340B Drug Pricing Program, which helps provide affordable medications to eligiblepatients. The program allows FQHCs to contract with local community pharmacies. Thisopportunity places community pharmacists in a unique position to care for underservedpatients, including monitoring medication adherence. Medication adherence plays animportant role in patients’ overall health. Non-‐adherence, which may be found in theform of prescription abandonment, may lead to increased hospitalizations, health carecosts, morbidity and mortality. An abandoned prescription is one that was filled by thepharmacy, but not picked up by a patient. Our focus will be to evaluate and compare therates and characteristics of abandoned prescriptions prescribed by FQHC providersversus all other non-‐FQHC providers at select 340B contracted community pharmacies.

Methods: Abandoned prescriptions at four 340B contracted community pharmacies, part of a grocery store-‐based chain within Ohio, will be identified during the studyperiod. The pharmacy database will be utilized to identify abandoned prescriptions andtheir characteristics such as whether it was prescribed by a FQHC provider or non-‐FQHC provider, the amount owed by the patient, and if it is a new or refilled prescription. Allprescriptions that are picked up (not abandoned) from the 340B contracted communitypharmacies will also be identified and their characteristics will be collected to use as acomparator group. Data will be analyzed once all information is collected.

Preliminary Results: Data collection will occur from February to May 2012. Preliminaryresults will be presented at The Ohio State University College of Pharmacy ResearchDay.

Conclusions: Study results will identify potential differences between groups and couldprovide opportunities to improve prescription abandonment rates in this FQHC patientpopulation.


Zinc Deficiency in the Context of ObesityEric Bolin, Shengying Bao, Mingjie Liu, Dara Burris, Daren KnoellDavis Heart and Lung Research Institute

Rank of First Author: UndergraduateProgram of First Author: Biochemistry

Obesity is a significant health care problem that causes increased morbidity andmortality worldwide. Zinc is known to play an essential role in preserving immunefunction whereas chronic obesity is associated with immune dysregulation resulting in alow-‐grade, systemic inflammatory state. Although zinc deficiency and obesity commonlycoexist in humans, it is not currently known what role, if any, that zinc plays in thesetting of obesity. Based on this, we investigated a mouse model of obesity todetermine whether zinc deficiency enhances the development of a chronicinflammatory state. To establish this we investigated obese and control mice that werefurther subject to zinc-‐deficient and zinc-‐sufficient dietary intakes. As expected, mice onhigh-‐fat diets gained significantly more weight (42%) when compared to control mice at10 weeks. Zinc deficient/high-‐fat fed mice exhibited a similar increase in fataccumulation and composition when compared to the obese control group. Thecombined zinc deficient/obese diet resulted increased NF-‐κB activity, a pro-‐inflammatory transcription factor, and decreased PPAR-‐γ activity, an anti-‐inflammatory transcription factor. Importantly, these changes, as determined by target geneactivation and a direct activity assay, were most pronounced in obese/zinc deficientmice. We are now examining whether changes in zinc metabolism at the cellular andtissue level modulate the activity of these proteins.Funding: Intramural competitive support provided by the OSU Food Innovation Center(DLK).


TREATMENT OF AUTOIMMUNITY THROUGH TOLERANCE

Deanna Brackman, Kristy Ainslie, Eric BachelderThe Ohio State University, College of Pharmacy, Division of Pharmaceutics

Rank of first author: UndergraduateProgram of first author: Pharmaceutical Sciences

Clinical studies have shown women with multiple sclerosis exhibit fewer to nosymptoms during pregnancy but have more frequent and severe symptoms in the firstfew postpartum months. During pregnancy the body is in what is known as a tolerogenicstate, meaning the immune system’s response to certain antigens is down-‐regulated. The aim of this study was to test drugs that mimic this tolerogenic state by decreasingimmune responses like nitric oxide release, while increasing proliferation of certain cellsubsets, such as FOXP3+ T regulatory cells. RAW 264.7 macrophages were exposed toestradiol, a hormone prevalent during pregnancy, and all-‐trans retinoic acid, acompound prevalent in the gut and vital during fetal development, to evaluate theirefficacy in suppressing an innate immune response. The nitric oxide (NO) production ofthe macrophages, when stimulated by lipopolysaccharides, was evaluated for bothcompounds. Our results indicated that NO levels were not altered by these compoundsand further analysis will need to be performed using other immune related cells.


PURIFICATION AND IDENTIFICATION OF AN ANTILEISHMANIALCOMPOUND FROM THE ROOTS OF THALICTRUM RUGOSUM

Anthony D. Gromovsky,1 C. Benjamin Naman,1 Gaurav Gupta,2 Claudio M. Lezama-‐Davila,2 Raymond W. Doskotch,1 Abhay R. Satoskar,2 and A. Douglas Kinghorn.11Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy and2Department of Pathology, College of Medicine, The Ohio State University, Columbus,Ohio 43210, USA

Rank of first author: UndergraduateProgram of first author: Division of Medicinal Chemistry and Pharmacognosy

Leishmaniasis is a significant public health concern throughout 88 countries worldwide,currently with approximately 12 million infected individuals.1 Transmitted by aninfected female sandfly vector, protozoan parasites of the genus Leishmania causeleishmaniasis via infiltration of tissue macrophages in host cells.2 Current treatmentsare known to be accompanied by adverse side effects, ranging from nausea to seriouscardiotoxicity.1 Isolation of endemic populations in developing countries has also stifledfunding for and efforts toward the discovery of more effective treatments, which isbecoming a growing concern as Leishmania species continue to develop increasing drugresistances to the most commonly employed treatments in modern medicine.1-‐3 Theneed for affordable, more effective, and less toxic treatments has led us to screensamples from a natural products library against L. donovani promastigotes. Sample tb-‐00097 displayed antileishmanial activity with IC50 = 1.12 µg/mL. This lead was observedto be impure and was purified chromatographically. Finally, the identity of tb-‐00097 was determined by NMR spectroscopy and mass spectrometry to be thalicthuberine,formerly isolated from the roots of Thalictrum rugosum.4

(1) Astelbauer, F.; Walochnik, J. Int. J. Antimicrob. Agents 2011, 38, 118-‐124.(2) Murray, H. W.; Berman, J. D.; Davies, C. R.; Saravia, N. G. Lancet 2005, 366, 1561-‐1577.(3) Croft, S. L.; Sundar, S.; Fairlamb, A. H. Clin. Microbiol. Rev. 2006, 19, 111-‐126.(4) Wu, W.-‐N.; Beal, J. L.; Doskotch, R. W. J. Nat. Prod. 1980, 43, 143–150.


SYNTHESIS AND CHARACTERIZATION OF ETHANOL-‐DEGRADING ACETALATED DEXTRAN POYLMER AND MICROPARTICLES

Kevin J. Kauffman1, Clement Do2, Sadhana Sharma2, Eric M. Bachelder2, Kristy M.Ainslie1,21William G. Lowrie Department of Chemical and Biomolecular Engineering, College ofEngineering2Division of Pharmaceutics, College of Pharmacy

Rank of first author: UndergraduateProgram of first author: Division of Pharmaceutics and Pharmaceutical Sciences

In the field of drug delivery, pH-‐sensitive polymeric microparticles can be used torelease therapeutic payloads slowly in extracellular conditions (pH 7.4) and faster inmore acidic areas in vivo, such as sites of inflammation, tumors, or intracellularconditions. Our group applies the pH-‐sensitive polymer acetalated dextran (Ac-‐DEX),which is a biodegradable polymer with highly tunable degradation kinetics. Ac-‐DEX hasdisplayed enhanced delivery of vaccine and drug components to immune and othercells, making it an extremely desirable polymer for immune applications. Currently, oneof the degradation products of Ac-‐DEX is methanol, which may cause toxicity issues ifapplied at high concentrations with repeated dosings. Therefore, in this project wereport the first synthesis and characterization of an Ac-‐DEX analog which instead has anethanol degradation product; we abbreviate this polymer as Ace-‐DEX, with the ‘e’ toindicate an ethanol byproduct. Like Ac-‐DEX, Ace-‐DEX microparticles have tunabledegradation rates at pH 5 (intracellular). These rates range from hours to several daysand are controlled simply by reaction time. Ace-‐DEX microparticles also showedminimal cytotoxicity compared to commonly-‐used poly(lactic-‐co-‐glycolic acid) (PLGA) microparticles when incubated with macrophages. This study aims to pave the way forthe use of Ace-‐DEX micro/nanoparticles in drug delivery and also allow acetalateddextran-‐type polymers to be used in high volume applications such as multiple dosingand tissue engineering.


ANTIPSYCHOTIC DRUG IMPACT ON DOPAMINERGIC NEURONS

Alex D. KlausingPharmaceutical Sciences


Antipsychotic drugs are used to treat schizophrenia. These drugs block dopamine D2

receptors and increase levels of the neurotransmitter dopamine in extracellularsignaling space (a measure of rate of dopamine utilization) to a similar extent. However,various antipsychotic drugs differ substantially in the amount of increase elicited in rateof synthesis of new dopamine molecules and in the amount of increase of dopamine’s metabolite, DOPAC (another measure of rate of dopamine utilization). Thus, twomarkers of dopamine utilization show divergent results when analyzing effects of someantipsychotic drugs. The purpose of this study is to explain this divergence. Acomputational model of dopamine varicosities was used. This model includes all knownmechanisms and pathways for dopamine and DOPAC creation, transport, andmetabolism. The activity of parameters in the model was varied until model outputmatched published experimental data. The results suggest that the effect of blockingdopamine D2 receptors (shared by all antipsychotic drugs) is to increase levels ofextracellular dopamine and to produce a small increase in rate of dopamine synthesisand in levels of DOPAC. Thus, these measurements are a valid indicator of rate ofdopamine utilization. The larger increases in rate of dopamine synthesis and in levels ofDOPAC observed after some drugs were best modeled by an increase in passivediffusion of dopamine from storage vesicles into cytosol. Review of the literaturedocuments that these drugs are all lipophilic weak bases which would be expected tosomewhat alkalinize storage vesicles. Rate of passive dopamine efflux is an expectedoutcome of vesicle alkalization. These findings suggest that the large increases in rate ofdopamine synthesis and in DOPAC level observed after some antipsychotic drugs results from physico-‐chemical properties interactions of the drugs with dopamine storagevesicles rather than from interaction with dopamine D2 receptors.

Acknowledgement to Lane Wallace


DEPLETION OF PIN1 IN MOUSE AORTIC ENDOTHELIAL CELLS INCREASESINDUCTION OF HYPOXIA-‐INDUCIBLE TRANSCRIPTION FACTORS

Abir J. Mneimneh, Dale G. HoytPharmacology


Oxygen is required in order for cells to survive. Consequently, cells havedeveloped mechanisms to adapt to changing oxygen concentrations. HypoxiaInducible Transcription factors 1 and 2 (HIF-‐1 and HIF-‐2) are induced byhypoxia. HIFs are believed to regulate expression of proteins that promotecell survival and function, allowing organisms to adapt to low oxygen.HIFs therefore may be manipulated to protect normal cells from hypoxia andor conceivably suffocate cancerous cells.

Our lab has investigated the regulation of signaling by PIN1, an enzymethat isomerizes phosphorylated serine/threonine-‐proline motifs inproteins. This activity allows PIN1 to modulate phosphorylation-‐dependentprotein functions. HIF-‐1 and 2 are regulated by phosphorylation, but it isnot yet known whether they are regulated by PIN1.

This question was addressed here. In order to investigate this we exposedmouse aortic endothelial cells, that either contained or lacked PIN1, to1% or 21% oxygen for 4 hours. Protein was then extracted from the cellsand western blotted to analyze for differences in the expression of HIF-‐1and 2.

Results. While the HIFs were not induced in endothelial cells containingPIN1 under these conditions, depletion of PIN1 increased the induction ofHIF-‐1 and HIF-‐2 by hypoxia 7-‐ and 5-‐fold, respectively.

The results indicate that PIN1 regulates induction of HIFs. Furthermore,they suggest that PIN1 may act on the phospho-‐serine/threonine-‐prolinemotifs in HIF proteins and/or components of the pathways regulating theHIFs in MAEC. Regulation of HIF expression in hypoxia by PIN1 could affectphysiological angiogenesis and/or tumor growth.


IMMUNOSTIMULATORY POLYSACCHARIDES AS A BASE MATERIAL FORPOLYMERIC MICROPARTICLES

Doug G. Montjoy1, Kevin J. Peine2, Eric M. Bachelder3, Kristy M. Ainslie1,31William G. Lowrie Department of Chemical and Biomolecular Engineering, College ofEngineering; 2Molecular Cell and Developmental Biology Graduate Program; 3College ofPharmacy Division of Pharmaceutics, The Ohio State University

Rank of first author: UndergraduateProgram of first author: Pharmaceutics

The use of polymers to enhance the delivery of vaccines is a growing area of research.Biocompatible polysaccharides such as dextran can be used as base materials to formacetalated polymers, such as acetalated dextran. Acetalated polymers have beenshown to have pH-‐sensitive characteristics ideal for delivery of vaccine elements toimmune cells. Microparticles formed from acetalated polysaccharides form vaccines byencapsulating a protein target (an antigen) and an immune danger signal (an adjuvant).Rather than adding an adjuvant to the vaccine system, immunostimulatory polymerscould be used in place of adding additional drug. To this end, different polysaccharidesare being tested to determine if they are immunostimulatory. The seven different polysaccharides that are being tested are laminarin, zymosan A, inulin, inulin (fromdahlia), glucan from euglena, glucan from barley, and curdlan. These polysaccharideshave been shown to activate immune cells and therefore are likely to prove to beimmunostimulatory. To evaluate these base materials, macrophages were incubatedwith four different concentrations of each polysaccharide (10 ug/ml, 1ug/ml, 100 ng/ml,or 10 ng/ml). As a positive control, lipopolysaccharide-‐stimulated (pro-‐inflammatory signal) macrophages were also studied as well as a negative control without treatment.The nitric oxide concentrations were evaluated in these test groups. Initial results showthat laminarin and inulin were shown to be immunostimulatory at 1 ug/ml. Further testing will be done on the other polysaccharides, and as well as laminarin and inulin, todetermine the immunostimulatory nature of the polysaccharides. After determining theimmunostimulatory nature and examining other properties of the polysaccharides, thepolysaccharides can be used as base materials for acetalation to generate a newpolymeric vaccine carrier.

The authors would like to thank Kevin Kauffman and Sadhana Sharma for their assistance.


EVALUATING DRUG THERAPY DECISION MAKING IN PATIENTS WITHEPILEPSY

James W. McAuley, PhD; Abbey M. Strazar, BS Pharm; Yun Jeong Lee, PharmD; Sheri L.Cotterman-‐Hart, MD, PhD; Bassel F. Shneker, MD, MBA

College of Pharmacy & Neurology Department, College of Medicine, The Ohio StateUniversity, Columbus Ohio

Rank of first author: UndergraduateProgram of first author: Pharmacy Practice and Administration

Background: When epilepsy patients are given advice about changes to their drugregimen, only a portion of them follow it. Decision making ultimately influencesadherence and non-‐adherence is dangerous for patients with epilepsy.Objective: The objective of this cross-‐sectional study was to determine why epilepsypatients do or do not follow suggested advice to change their drug therapy.Methods: Four weeks after their clinic visit, 100 patients were sent a survey askingthem about the prescriber-‐recommended drug regimen changes, whether they followedthe advice, and the main reasons for their decisions.Results: Of the fifty-‐one responses received, nearly all (94%) reported that they didfollow the suggested advice for changes in their drug regimen. Their reasons includedthe desire to have fewer seizures (46%), less side effects (17%) and “I trust mypractitioner” (26%).Conclusions: Beyond the desire to have less seizures & side effects, patients report“being heard by” and “trust in” their epilepsy specialist as significantly influencing theirdecision making regarding changes to their drug therapy.


SYNTHESIS OF ALLOSTERIC MODULATORS FOR NICOTINIC ACETYLCHOLINE RECEPTORS

Jason Young,1 Bitna Yi,2 Tatiana Gonzalez-‐Muskus,2 Dennis McKay,2 and Karl Werbovetz11Division of Medicinal Chemistry and Pharmacognosy and 2Division of Pharmacology,College of Pharmacy, The Ohio State University, Columbus, OH

Rank of first author: UndergraduateProgram of first author: Division of Medicinal Chemistry and Pharmacognosy

Nicotinic acetylcholine receptors (nAChR) can be found throughout the human nervoussystem. The receptors regulate a multitude of functions, including development,inflammation, and movement. They also serve as the receptor site for nicotine, anextremely addictive drug. Novel therapeutic strategies for breaking this addictioninvolve synthesis of negative allosteric modulators that could deactivate the binding sitefor nicotine on these receptors. However, because many subtypes of the nAChR exist, itis difficult to target one without affecting others. This study aims to synthesize a seriesof analogs of compound 16, an arylsulfonyl piperazine-‐containing compound that waspreviously shown to display selectivity for the Hα4β2 nAChR compared to the Hα3β4nAChR receptor subtype.1 Synthetic pathways are focused on amide bond formationbetween substituted arylsulfonyl piperazines and aryl amines. To date, nine derivativesof 16 have been synthesized. Current results confirm the identity of these compoundsby 1H, 13C,m/z, and elemental analyses. Biological testing of these compounds has, thusfar, shown that the analogs retain the potency of 16 for the Hα4β2 nAChR, but have lostselectivity for that receptor subtype. Future work will focus on exploring differenthypotheses regarding the basis of 16 receptor subtype selectivity through the synthesis and evaluation of additional analogs.

1. Henderson, B.; Carper, D.; González-‐Cestari, T.; Yi, B.; Mahasenan, K.; Pavlovicz, R.; Dalefield, M.;Coleman, R.; Li, C.; McKay, D. J. Med. Chem. 2011, 54, 8681.


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