ŷƋƈŃƉƘƗƘƕƈŃƐƌƖƖƌƒƑŃƒƉŃƓƋƄƕƐƄƆƈƘƗƌƆƄƏŃƖƆƌƈƑƆƈƖŃƗƋƄƗŃƐƌƕƕƒƕƖŃƗƋƈŃ ƋƌƖƗƒƕƜŃƒƉŃƗƋƈŃƓƕƒƊƕƈƖƖŃƒƉŃƐƄƑƎƌƑƇŃŃ
ؙ7KHVWDUWRIWKHUHODWLRQVKLSEHWZHHQKXPDQLW\DQGPHGLFLQHVFDQEHWUDFHG EDFNWRGLVWDQWDQFLHQWKLVWRU\%HJLQQLQJWKRXVDQGVRI\HDUVDJRKXPDQVKDYH VHOHFWHGDQGXVHGQDWXUDOPHGLFLQHVGHULYHGIURPDQLPDOVSODQWVDQGPLQHUDOV RQDQH[SHULPHQWDOEDVLV:LWKWKHVWDUWOLQJDGYDQFHVLQrQHFKHPLVWU\PDGH GXULQJ WKH WK FHQWXU\ GUDPDWLF DGYDQFHV ZHUH PDGH LQ GUXJ V\QWKHVLV WHFKQRORJLHVDQGQHZSKDUPDFHXWLFDOVZHUHGHYHORSHGRQHDIWHUDQRWKHU:LWK WKHVWFHQWXU\SRVLWLRQHGWREHDQDJHRIOLIHVFLHQFHVLWLVH[SHFWHGWKDW WKHUHZLOOEHJUHDWRSSRUWXQLWLHVWRDFKLHYHIXUWKHUGUDPDWLFGHYHORSPHQWDQG H[SDQVLRQLQWKHr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tGLVWXUEDQFHVLQWKHKHDOWKRIWKHJOREDO HFRV\VWHP ZKLFK LQFOXGHV WKH KXPDQ VSHFLHV 7KH IRFXV LV RQ WKH UROH RI SKDUPDF\ DV D FRPSUHKHQVLYH KHDOWK VFLHQFH DV D PHDQV RI UHVROYLQJ WKHVH SUREOHPV ŤؙƐƈƖƖƄƊƈؙƗƒؙƗƋƒƖƈؙƌƑƗƈƑƇƌƑƊؙƗƒؙƖƗƘƇƜؙƓƋƄƕƐƄƆƈƘƗƌƆƄƏŃ ƖƆƌƈƑƆƈƖŃƗƒŃƓƕƒƙƌƇƈŃƄŃƅƈƗƗƈƕŃƉƘƗƘƕƈŃƉƒƕŃƄƏƏŃƏƌƙƌƑƊŃƗƋƌƑƊƖ
ؙ3KDUPDFHXWLFDOVFLHQFHVLVDQDFDGHPLFGLVFLSOLQHZLWKWKHPLVVLRQRIKHOSLQJ WRGHYHORSDERXQWLIXOVRFLHW\WKURXJKFRPSUHKHQVLYHVFLHQWLrFUHVHDUFKLQWRWKH FKHPLFDOVXEVWDQFHVZKLFKDUHUHODWHGWRDOONLQGVRIOLIHRQHDUWKDQGE\ FRQWULEXWLQJWRWKHKHDOWKRIKXPDQLW\WKURXJKWKHFUHDWLRQRISKDUPDFHXWLFDOV &UXFLDO WR IXUWKHU DGYDQFHV LQ SKDUPDF\ LV \RXWKIXO VWUHQJWK ZLWK DPSOH FUHDWLYLW\DQGERXQGOHVVDPELWLRQ:HQHHGSHUVRQVZKRFDQXVHWKDWVWUHQJWK DQG FRQWULEXWH WR WKH ZHOIDUH RI KXPDQLW\ ZKLOH IXOrOOLQJ WKH PLVVLRQ RI SKDUPDF\ RQ WKH JOREDO VWDJH 7KLV LV WKH W\SH RI SHUVRQV ZKR VKRXOG EH UHVSRQVLEOHIRUQHZGHYHORSPHQWLQOLIHVFLHQFHVDQGPHGLFLQDOVFLHQFHVLQWKH VWFHQWXU\ ؙ$UH\RXLQWHUHVWHGLQWDNLQJWKHrUVWVWHSVLQWKDWGLUHFWLRQDWWKH*UDGXDWH "\6FKRROؙDQGؙ6FKRROؙRIؙ3KDUPDFHXWLFDOؙ6FLHQFHVؙDWؙ2VDNDؙ8QLYHUVLW &RPSUHKHQVLYHDQGHQJDJLQJHGXFDWLRQDODQGUHVHDUFKrHOGVDUHZDLWLQJIRU\RX DW2VDND8QLYHUVLW\:HVWURQJO\KRSH\RXZLOOVKDUH\RXUGUHDPVZLWKXVOHDUQ ZLWKXVDQGSXUVXHUHVHDUFKWR\RXUKHDUW VFRQWHQWV
3URI Experimental InsƟtute for Medicinal Plants Clinical Pharmacy Unit Center for Advanced ĚƵĐĂƟon and Research in Pharm.D EduĐĂƟon Unit PharmaceƵƟcal Sciences Advanced PharmaceƵƟcal Sciences Unit Technology InnovaƟon Unit Drug InnovaƟon Center Clinical Drug Development Unit ƌĞĂƟǀĞ Research Unit Unit for PharmaceƵƟcal ĚƵĐĂƟon in Community Medical Care Unit for PromŽƟŽn of Community Self- MediĐĂƟon Center for PharmaceƵƟcal Unit for PharmaceƵƟcal Research in Community EducĂƟon and Research in Medical Care Community Medical Care Unit for PharmaceƵƟcal ĚƵĐĂƟon in Hospital Community Medical Care Unit for PharmaceƵƟcal ĚƵĐĂƟon in Pharmacy Community Medical Care Cell Biology(1) Biomolecular Science and Engineering(1) Department of Medical InnovaƟon(2) Clinical PharmaceƵƟcals(2) Experimental Genome Research(3) ŽŽƉĞƌĂƟǀĞ Fields / CoopeƌĂƟŶŐ Graduate School CollaborĂƟŽŶ Nano-Design for InnoǀĂƟǀĞ Drug Development(3) (1)The /ŶƐƟtute of ScieŶƟĮc and Industrial Applied Pharmacognosy(4) Research Molecular pathophysiology(5) (2)Osaka University Hospital Department of Pharmacy Biomedical InnovaƟon(6) (3)Research /ŶƐƟtute for Microbial Diseases Clinical and Experimental Pathophysiology(7) (4)The Museum of Osaka University (5)NaƟonal Cerebral and Cardiovascular Center PharmaceƵƟcal and Medical Device Regulatory(8) (6)NaƟonal /ŶƐƟƚƵƚes of Biomedical InnoǀĂƟon, Regulatory Science(9) Health and NutriƟon (7)Osaka Medical Center for Cancer and PreempƟve Medical Pharmacology for Mind and Endowed Chair Cardiovascular Diseases Body (8) PharmaceƵƟĐĂůƐ and Medical Devices Agency (9) NĂƟŽŶĂl /ŶƐƟtute of Health Sciences Joint Research Chair Advanced CosmeƟc Science Laboratory of Biophysical Chemistry Professor Tadayasu OHKUBO +81-6-6879-8220 [email protected] Associate Professor Takuya YOSHIDA +81-6-6879-8222 [email protected] Specially Appointed Assistant Professor Kazuki KAWAHARA +81-6-6879-8222 [email protected] FAX +81-6-6879-8222 Professor Tadayasu OHKUBO In this laboratory, we aim to determine three-dimensional structures of proteins and their complexes in order to obtain novel structural insights into their functions. X-ray crystallography and NMR spectroscopy are conducted to define molecular structure at an atomic level. Intermolecular interactions, structural stability, complex formation, intramolecular dynamics and reaction kinetics are probed with various physico-chemical techniques including spectroscopic, hydrodynamic and thermodynamic analy- ses, i.e. circular dichroism, analytical ultra centrifugation, isothermal titration calorimetry, etc. Furthermore, mass spectroscopy is also used to identify interacting partners and to characterize post-translational modifications and structural integrity of target proteins. Protein engineering and chemical syntheses are used in sample preparations. The structures of protein molecules under study at our laboratory are wide-ranging. They include mammalian disease-related proteins such as Lipocalin-type Prostaglandin D Synthase (L-PGDS), Receptor for Advanced Glycation Endproducts (RAGE), 3HUR[LVRPH3UROLIHUDWRU$FWLYDWHG5HFHSWRUȖ 33$5Ȗ 3LJPHQW(SLWKHOLXP'HULYHG)DFWRU 3(') DQGSURWHLQVIURPSDWKR- genic bacteria. We utilize an integrated approach spanning biophysics, biochemistry, chemical biology and structural biology with the aim of understanding of the mechanisms regulating biological functions of these proteins and their complexes. The experimen- tally derived data are combined with molecular dynamics simulations to provide functional models and venues of structure-based drug discovery (SBDD). Research topics 1) Structure determination of proteins with X-ray crystallography and NMR spectroscopy 2) Physico-chemical analysis on molecular properties and interactions of proteins 3) Development of protein production systems for physico-chemical studies 4) Computational approach for protein dynamics analysis and drug discovery Recent publications 1) Kawahara K et al., Homo-trimeric Structure of the Type IVb Minor Pilin CofB Suggests Mechanism of CFA/III Pilus Assem- bly in Human Enterotoxigenic Escherichia coli. J Mol Biol., 428, 1209-1226, 2016. 2) Takuwa A et al., Ordered self-assembly of the collagenous domain of adiponectin with noncovalent interactions via glycosyl- ated lysine residues. FEBS Lett., 590, 195-201, 2016. 3) Qin S et al., Thermodynamic and NMR analyses of NADPH binding to lipocalin-type prostaglandin D synthase. Biochem Biophys Res Commun. 468, 234-239, 2015. 4) Kawahara K et al., Cloning, expression, purification, crystallization and X-ray crystallographic analysis of CofB, the minor pilin subunit of CFA/III from human enterotoxigenic Escherichia coli. Acta Crystallogr F Struct Biol Commun. 71, 663-667, 2015. +DUDGD6HWDO6WUXFWXUDOEDVLVIRU33$5ȖWUDQVDFWLYDWLRQE\HQGRFULQHGLVUXSWLQJRUJDQRWLQFRPSRXQGV6FL5HS 2015. 1 Laboratory of Synthetic Medicinal Chemistry Professor Shuji AKAI +81-6-6879-8210 [email protected] Associate Professor Takashi IKAWA +81-6-6879-8212 [email protected] Professor Shuji AKAI Most of active substances of medicines are functional organic molecules, both naturally occurring and man-made, and complex biological processes in living organisms are accurately controlled by chemical interplay of organic molecules at the molecular level. In this sense, organic chemistry has been playing critical roles for drug discovery. Nowadays, the development of environ- mentally benign methods for producing medicines is another urgent challenge. As a means to accommodate these requests, our research group has been engaged in (1) the syntheses of biologically important natural products and their derivatives as well as artificial molecules which are expected to become drug candidates and (2) the creation of new synthetic methods, strategies and catalysts. Some of the noteworthy projects and results are as follows: We have been studying total syntheses of bioactive natural products, such as allocolchicine, aloin, viridin, and their derivatives. The purpose of our total syntheses involves not only creating a process for retrieving a large supply of target compounds but also designing new functional molecules that would serve as potent leads for future drug discovery. For this purpose, the known synthetic methods are not always satisfactory, and therefore, we also have focused our efforts on devising new methods to find solutions by best employing characteristic features of typical elements, transition metals, enzymes, etc. A typical example is given by the lipase/oxovanadium combo-catalyzed dynamic kinetic resolution that quantitatively converts racemic alcohols into optically pure molecules. Effective utilization of very reactive species, such as benzynes, as key synthetic intermediates, is 7RWDO6\QWKHVLVRI 'HYHORSPHQWRI1HZ our other big projects to construct complex molecules in %LRDFWLYH0ROHFXOHV 6\QWKHWLF0HWKRGV F MeO OMe single steps. We have also developed methods for regioselec- MeO tive fluorination of benzenes to effectively produce fluorinated MeO fluorinated molecules which are of particular interest as drug candidates. OH O OH NHAc NSC51046 Interdisciplinary research is also currently underway HO OH HQ]\PDWLFG\QDPLFNLQHWLFUHVROXWLRQ HO H MeO Me Me O OH OMe through cooperation with other research groups, pursuing imperanene Me Me HO OH OO pharmacological evaluations of the synthesized compounds OH OH aloin B O N O as well as gaining vital clue to understanding of their biologi- O N O H O cal functions. O H H N O R (–)-crinane N N H H O hippadine himbacine analog Research topics 1) Total synthesis of biologically important natural products 2) Development of new asymmetric syntheses using biocatalysts 3) Control of benzyne reactions and their synthetic applications 4) Creation of new functional molecules for drug discovery and medical treatments Recent publications 1) S. Akai et al., Preparation of Optically Active Cycloalkenes Bearing All-carbon Quaternary Stereogenic Centres via Lipase- oxovanadium Combo-catalysed Dynamic Kinetic Resolution. Green Chem., in press, 2016. 2) T. Ikawa et al., 1,3- and 1,4-Benzdiyne Equivalents for Regioselective Synthesis of Polycyclic Heterocycles. Chem. Sci., 7, 5206, 2016. 3) S. Akai et al., Diversity Oriented Synthesis of Allocolchicinoids with Fluoro and/or Oxygen Substituent(s) on the C-Ring from a Single Common Intermediate. Eur. J. Org. Chem., 1562, 2016. 4) T. Ikawa et al., 2-[(Neopentyl glycolato)boryl]phenyl Triflates and Halides for Fluoride-Ion-Mediated Generation of Function- alized Benzynes. Adv. Synth. Cat., 357, 2287, 2015. 5) S. Akai et al., A Mesoporous-Silica-Immobilized Oxovanadium Cocatalyst for the Lipase-Catalyzed Dynamic Kinetic Resolu- tion of Racemic Alcohols. Angew. Chem. Int. Ed., 52, 3654, 2013. 6) T. Ikawa et al., ortho-Selective Nucleophilic Addition of Primary Amines to Silylbenzynes: Synthesis of 2-Silylanilines. Angew. Chem. Int. Ed., 50, 5674, 2011. 7) T. Ikawa et al., Preparation and Regioselective Diels–Alder Reactions of Borylbenzynes: Synthesis of Functionalized Arylbo- ronates. Angew. Chem. Int. Ed., 49, 5563, 2010. 2 Laboratory of Bio-Functional Molecular Chemistry Professor Kiyohito YAGI +81-6-6879-8195 [email protected] Assistant Professor Akihiro WATARI +81-6-6879-8198 [email protected] FAX +81-6-6879-8199 Professor Kiyohito YAGI Multicellular organisms contain various compositionally distinct fluid compartments, which are established by epithelial and endothelial cell sheets. Recent findings show that tight junctions (TJs) play pivotal roles in barrier and solutes movement in these cell sheets, resulting in creation and maintenance of these compartments. TJ is one of the intercellular junctional complexes in epithelial cells at the apical compartment of the lateral membrane. TJs have two important roles, barrier and fence functions. Modulation of TJ-barrier is a promising strategy for drug delivery. TJs control cellular polarity by prevention of free movement of transports and receptors on the membrane. Dysfunction of TJs causes disruption of the cellular polarity, leading to oncogenic transformation. Current evidence suggests that a component of TJ is a co-receptor for hepatitis C virus. These findings on TJs provide us new insight into drug delivery, anticancer and antiviral therapies, but little attention has been paid to potential druggable targets in TJs. We are trying to develop and propose a novel concept for pharmaceutical research, the TJ-based drug discovery. Our laboratory has been conducting basic research on the modulation of cell function for medical and biotechnological applications. We had tried to develop novel therapy for acute and chronic liver diseases using Tissue Engineering. Recently we isolated a novel stem cell (TGPCs) from wisdom teeth. TGPCs differentiated into cells of three germ layers including osteoblasts, neural cells, and hepatocytes, and intravenous injection of TGPCs alleviated hepatic failure in CCl4-treated rats, suggesting that TGPC is a promising candidate for cell-based therapy. We will develop TGPC-based cell therapy and apply TGPCs-derived hepatocytes into predicting action of drug candidates. We also aim to evaluate safety of nano-sized materials and to construct biosensor using microbial carotenoid synthesizing system to detect very small amount of substances. Research topics 1) Development of novel therapeutic strategies for hepatitis C virus 2) Cell-based pharmaceutical research 3) Tight junction-based drug discovery 4) Nanotoxicology 5) Development of biosensor using carotenoid biosynthesis system in photosynthetic bacteria Recent publications 1) Watari A., et al., Checkpoint Kinase 1 Activation Enhances Intestinal Epithelial Barrier Function via Regulation of Claudin-5 Expression. PLoS One, 11(1):e0145631, 2016. 2) Suzuki H., et al., C-terminal Clostridium perfringens enterotoxin is an efficient nasal vaccine delivery system for pneumococ- cal vaccine. PLos one, 10(5) :e0126352, 2015. 3) Matsuhisa K., et al., Anti-HCV effect of mycelia solid culture extracts of Lentinula edodes and low-molecular-weight lignin. BBRC, 462(1):52-7, 2015. 4) Fukasawa M., et al. Monoclonal antibodies against extracellular domains of claudin-1 block hepatitis C virus infection in a mouse model. J Virol. 89(9):4866-79. 2015. 5) Watari A., et al., Homoharringtonine increases intestinal epithelial permeability by modulating specific claudin isoforms in Caco-2 cell monolayers. Eur J Pharm Biopharm., 89:232-8, 2015. 6) Li X., et al., Development of an anti-claudin-3 and -4 bispecific monoclonal antibody for cancer diagnosis and therapy. J Phar- macol Exp Ther., 351(1), 206-13, 2014. 7) Takahashi A., et al., Creation and biochemical analysis of a broad-specific claudin binder. Biomaterials, 33, 3464-3474, 2012. 8) Kondoh M., et al., Spiral progression in the development of absorption enhancers based on the biology of tight junctions. Adv. Drug Deliv. Rev.,17, 515-522, 2012. 3 Laboratory of Biochemistry and Molecular Biology Professor Hiroyuki MIZUGUCHI +81-6-6879-8185 [email protected] Associate Professor Fuminori SAKURAI +81-6-6879-8188 [email protected] Specially Appointed Assistant Professor Eiko SAKAI +81-6-6879-8187 [email protected] Specially Appointed Assistant Professor Kazuo TAKAYAMA +81-6-6879-8187 [email protected] FAX +81-6-6879-8186 Professor Hiroyuki MIZUGUCHI For the elucidation of the functions of genes and proteins, conventional Ad vec to r Ad type 35 AdRGD AdK7 Ad TAT AdF35 which is the first stage of biological drug development, (Ad type 5 vector) vector comprehensive genomic and proteomic research has been carried out. Much of this research, however, only allows the estimation of gene functions, and is insufficient for the final verification of a gene’s mechanisms of action and the poten- tial applications of genomic information to drug develop- ment and medical technology. Therefore, it is necessary to empirically analyze the functions of candidate genes and proteins in detail. An important approach to this empirical CAR CAR CAR analysis is to examine the functions of genes and proteins at He p aran CAR CD46 v-integrin the levels of the cell and the whole body by transfecting the sulfate candidate gene or a family of genes into target cells. Our laboratory develops fundamental technologies that permit Characteristics of gene delivery by various types of highly efficient gene transfer, high levels of expression in target cells, tissue-tropic gene transfer, and the control of capsid-modified adenovirus (Ad) vectors gene and protein expression. By taking advantage of the characteristics of the adenovirus vector as a gene transfer vector (highly efficient gene transfer activity, the ability to produce a high titer of the vector, simple and easy production of the vector, and potential for in vivo applications), we develop highly effective and versatile next-generation gene transfer vectors. These vector systems are essential for the gene function studies. Gene transfer technologies also serve as a foundation for the development of highly effective and safe vaccines and vectors for gene therapy and genetically modified cell therapy (regenerative medicine). Thus, this gene transfer technology can be applied to a wide range of areas. Our laboratory also researches on molecular biological studies about the non-coding RNA (microRNA) for the cell function, the stem-cell biologi- cal studies for drug screening and regenerative medicine, and the immunological studies against adenovirus vectors. For example, hepatocyte- like cells and enterocyte-like cells differentiated from human induced pluripotent stem cells are expected to be utilized as a tool for drug screen- ing. All these studies are carried out by using the next-generation gene transfer vectors we developed. Research topics 1) Development of new gene transfer vectors and their application to gene therapy, vaccines, and regenerative medicine 2) Research on non-coding RNA (microRNA) for the cell function 3) Research on stem-cell biology for drug screening and regenerative medicine 4) Analysis of the molecular mechanism on immune response against viral vectors and the development of safer vectors Recent publications 1) Nagamoto Y. et al., Enhancement of survival rate by human iPSC-derived hepatocyte sheet transplantation in acute liver failure mice. J. Hepatol., 64, 1068-1075 (2016) 2) Sakurai F. et al., Efficient detection of human circulating tumor cells without significant production of false-positive cells by a novel conditionally replicating adenovirus. Mol.Ther.Methods.Clin.Dev., 3:16001 (2016) 3) Machitani M. et al.,NF-kB promotes leaky expression of adenovirus genes in a replication-incompetent adenovirus vector. Sci. Rep., 6:19922 (2016) 4) Tsuzuki S. et al., TANK-binding kinase 1-dependent or -independent signaling elicits the cell type-specific innate immune responses induced by the adenovirus vector. Int. Immunol., 28, 105-115 (2016) 5) Ozawa T S. et al., Generation of enterocyte-like cells from human induced pluripotent stem cells for drug absorption and metabolism studies in human small intestine. Sci. Rep., 5:16479 (2015) 6) Takayama K. et al., Prediction of inter-individual differences in hepatic functions and drug sensitivity by using human iPS-derived hepatocytes. Proc. Natl. Acad. Sci. USA, 111, 16772-16777 (2014) 7) Blumberg R. et al., Protective mucosal immunity mediated by epithelial CD1d and IL-10, Nature, 509, 497-502(2014) 7DND\DPD.HWDO&&$$7HQKDQFHUELQGLQJSURWHLQPHGLDWHGUHJXODWLRQRI7*)ȕUHFHSWRUH[SUHVVLRQGHWHUPLQHWKHKHSDWREODVW fate decision. Development, 141, 91-100(2014) 9) Takayama K. et al., Long-term self-renewal of human ES/iPS-derived hepatoblast-like cells on human Laminin 111-coated dishes. Stem Cell Rep., 1，322-335 (2013) 10)Sugio K. et al., Enhanced safety profiles of the telomerase-specific replication-competent adenovirus (Telomelysin) by incorporation of normal cell-specific microRNA-targeted sequences. Clin. Cancer Res., 11)Yamaguchi T. et al., Induction of type I interferon by adenovirus-encoded small RNAs. Proc. Natl. Acad. Sci. USA, (2010) 4 Laboratory of Synthetic Organic Chemistry Professor Hiromichi FUJIOKA +81-6-6879-8225 [email protected] Associate Professor Mitsuhiro ARISAWA +81-6-6879-8226 [email protected] Assistant Professor Kenichi MURAI +81-6-6879-8227 [email protected] FAX +81-6-6879-8229 Professor Hiromichi FUJIOKA Since the ancient times, people have been using natural plants and minerals as remedies to help curing diseases. In the course of the 20th Century, accompanying the progress of modern science, the active principles contained in these substances has been identified, the chemical structures of the latter have been determined, and their production by industrial organic chemistry has become possible. We immediately entered an era of sulfa drugs and antibiotics, with a rapid improvement in the chemistry of medicinal drugs. Now that we have stepped into the 21st Century, the genome information is being collected, and coupled with the improvement of computers, an era where the needed drugs can be at last designed and synthesized is about to begin. Synthetic organic chemistry is a discipline related to the creation of medicinal drugs and related compounds using methods from organic chemistry. Our field has the objective to train exceptional synthetic organic chemists who are to be experts in the development of new drugs, and we are involved in research with the aim to discover quality medicinal drugs. In particular, the research is approached with interest in using organic synthesis to massively supply biologically active molecules that are naturally present in small amounts and therefore difficult to extract. It is another important objective to perform research to develop new organic reactions and reagents allowing the synthesis of these molecules to be completed, and to elucidate the reaction mechanism, thereby contributing purely to the progress of organic chemistry itself. In such synthetic processes, based on information from every field of pharmaceutical science, it will be also possible to design and synthesize useful drugs that can outperform the ones that nature provided. The establishment of new convenient methods for the synthesis of peri-hydroxy polycyclic aromatic compounds, the development of highly specific asymmetric reactions and remote asymmetric induction reactions that proceed via reactive cations, the development of new reactions using ketene acetal type compounds and asymmetric catalysts are some results of our recent research work. By applying these basic techniques, biologically active natural products were synthesized in short steps by asymmetric synthesis. In addition, while applying these results, research is underway to synthesize biological inhibitors that are conceived based on the information on their receptors with the use of computer-aided molecular design. We are also actively involved in the development of environment-friendly catalytic organic reactions. Research topics 1) Development and application of novel organic reactions and reagents 2) Development of asymmetric synthetic reactions that employ reactive chemicals 3) Molecular design and synthetic chemistry for the development of medicinal drugs 4) Synthesis of antitumor antibiotics and related antitumor agents 5) Development of environment-conscious reactions Recent publications 1) Arisawa M. et al., Redox Switching of Orthoquinone-Containing Aromatic Compounds using Gas Energy. Angew. Chem. Int. Ed., 55, 7432–7436, 2016. 2) Murai K. et al., Oxidative Rearrangement via in Situ Generated N-Chloroamine: Synthesis of Fused Tetrahydroisoquinolines. Org. Lett., 18, 1314–1317, 2016. 3) Fujioka H. et al., Stereoselective Construction of 2,7-Disubstituted fused-Bis Tetrahydrofuran Skeletons: Biomimetic-Type 6\QWKHVLVDQG%LRORJLFDO(YDOXDWLRQRI DQG í $SO\VLDOOHQHDQG7KHLU'HULYDWLYHVJ. Org. Chem. 80, 10261–10277, 2015. 4) Arisawa M. et al., Development of Enyne Metathesis/Isomerization/Diels-Alder One-pot Reaction for Novel Near-Infrared (NIR) Dye Core. Chem. Eur. J. 23, 17491-17494, 2015. 5) Murai K. et al., Enantioselective iodolactonization of allenoic acids. Chem. Commun. 50, 12530-12533, 2014. 5 Laboratory of Analytical Chemistry Professor Tadayuki UNO +81-6-6879-8205 [email protected] Assistant Professor Hirofumi TSUJINO +81-6-6879-8208 [email protected] FAX +81-6-6879-8209 Professor Tadayuki UNO Oxygen is a molecule that is indispensable to the life of human beings, and biological reactions of the oxygen are mainly regulated by heme proteins. Heme protein is a generic name of proteins which contain heme prosthetic groups, and plays a key role not only in oxidative metabolism of various drugs and chemical compounds, but also in signal transduction in response to environmental changes. In our Laboratory of Analytical Chemistry, we study structure-function relationships of heme proteins which are responsible for oxygen metabolism in our body. We hope to reveal biological events which are related to oxygen, and to contribute greatly to the field of drug discovery and clinical usage of drugs. For example, neuroglobin is induced in our brain under hypoxia and protects neurons by releasing oxygen, and nitrophorin releases nitrogen oxide (NO) and expands blood vessels. Understanding of the gas-storage/release mechanisms of these heme proteins will lead to a development of neuro-protective and cardiovascular drugs, respectively. On the other hand, cytochrome P450 (CYP) is expressed mainly in our liver and is a key protein in drug metabolism. Understanding of the drug metabolizing mecha- nism of CYP will enable safer drug usage for the patients with low side effects. We cloned and over-expressed almost all of human major CYPs, and are analyzing their molecular structure using spectroscopic methods such as UV-vis absorption and resonance Raman effect. Their drug binding and metabolizing activities are also measured with HPLC and MS technique, revealing structure- function relationships of clinically important human CYPs. Our research project is situated in the interdisciplinary field of analytical chemistry, physical chemistry, inorganic chemistry, and molecular biology. We afford an environment for learning research mind with high originality, fully aided by wide knowledge of the interdisciplinary field. We educate students who are expected to be distinguished researchers and can perform research from various points of view. We welcome students from all over the world who are Crystal Structure of Human Drug Metabolizing Enzyme interested in our research field. CYP2C9 Interacting with Warfarin (PDB code: 1OG5). Research topics 1) Structure of globin proteins and reaction with reactive oxygen species 2) Engineering of gas-carrier proteins based on recombinant DNA technique 3) Drug recognition and metabolizing properties of human CYPs and effect of SNPs 4) Oxygen activation mechanism of heme proteins as revealed by resonance Raman spectroscopy Recent publications 1) Miyamoto M. et al., Membrane anchor of cytochrome P450 reductase suppresses the uncoupling of cytochrome P450. Chem. Pharm. Bull. 63, 286-294, 2015 2) Li X. et al., Development of an anti-claudin-3 and -4 bispecific monoclonal antibody for cancer diagnosis and therapy. J. Pharmacol. Exp. Ther. 351, 206-213, 2014 3) Yamashita T. et al., Ferric Human Neuroglobin Scavenges Superoxide to Form Oxy Adduct. Chem. Pharm. Bull. 62, 613-615, 2014 4) Tsujino H. et al., Disulfide Bonds Regulate Binding of Exogenous Ligand to Human Cytoglobin. J. Inorg. Biochem. 135, 20-27, 2014 5) Tamer ZA. et al., Effect of Cytochrome P450 2C19 and 2C9 Amino Acid Residues 72 and 241 on Metabolism of Tricyclic Antidepressant Drugs. Chem. Pharm. Bull. 62, 176-181, 2014 6 Laboratory of Bioorganic Chemistry Professor Satoshi OBIKA +81-6-6879-8200 [email protected] Assistant Professor Tsuyoshi YAMAMOTO +81-6-6879-8202 [email protected] Specially Appointed Assistant Professor Osamu NAKAGAWA +81-6-6879-8202 [email protected] FAX +81-6-6879-8204 Professor Satoshi OBIKA The research interests of our laboratory are focused on chemical events in living system. On the basis of biomolecules such as nucleic acids, sugars, and other natural products, we are engaged in design and syntheses of novel artificial molecules having exquisite biologi- cal functions. We also investigate their functions in order to reveal the mechanism of action and to develop more efficient molecules. Antisense and antigene strategies for selective regulation of gene expression by appropriate oligonucleotide analogues, as a direct way to treat serious diseases such as cancer and viral infec- tions, are potential strategies for rational drug discovery. However, due to lack of metabolic stability of natural oligonucleotides under physiological conditions, the successful implementation of antisense and antigene strategies largely depends on the availability of chemi- cally modified oligonucleotides. We achieved the first synthesis of a novel nucleoside with a fixed N-type conformation, 2'-O,4'- C-methylene bridged nucleic acid (2',4'-BNA/LNA) and found that the 2',4'-BNA/LNA modified oligonucleotides could be a promising candidate for practical antisense and antigene strate- gies. Furthermore, we have been developing a lot number of nucleic acid analogues for application to several genome technologies, e.g., RNA interference (RNAi) and detection of single nucleotide polymorphisms (SNPs) or a damaged DNA. Drug delivery systems based on the oligonucleotide conjugates are also studied for practical application of therapeutic oligo- nucleotides. Some of the research programs described above are progressed in cooperation with various research institutes. Research topics 1) Novel nucleic acid analogs - design, synthesis and biofunction 2) Application of the novel nucleic acid analogues to genome technology 3) Chemistry and biology of oligonucleotide conjugates Recent publications 1) K. Morihiro, T. Kodama, S. Tsunoda, S. Obika, Wavelength-selective light-triggered strand exchange reaction. Org. Biomol. Chem., 2016, 14, 1555-1558. 2) S. Wada, H. Yasuhara, F. Wada, M. Sawamura, R. Waki, T. Yamamoto, M. Harada-Shiba, S. Obika, Evaluation of the effects of chemically different linkers on the hepatic accumulation, cell tropism and gene silencing ability of cholesterol-conjugated antisense oligonucleotides. J. Controlled Release, 2016, 226, 57-65. 3) T. Yamamoto, M. Sawamura, F. Wada, M. Harada-Shiba, S. Obika, Serial incorporation of a monovalent GalNAc phosphora- midite unit into hepatocyte-targeting antisense oligonucleotides. Bioorg. Med. Chem., 2016, 24, 26-32. 4) H. Hoshino, Y. Kasahara, H. Fujita, M. Kuwahara, K. Morihiro, S. Tsunoda, S. Obika, Consecutive incorporation of function- alized nucleotides with amphiphilic side chains by novel KOD polymerase mutant. Bioorg. Med. Chem. Lett., 2016, 26, 530-533. 5) T. Osawa, Y. Hari, M. Dohi, Y. Matsuda, S. Obika, Synthesis and properties of the 5-methyluridine derivative of 3,4-dihydro- 2H-pyran-bridged nucleic acid (DpNA). J. Org. Chem., 2015, 80, 10474-10481. 6) S. Hori, T. Yamamoto, R. Waki, S. Wada, F. Wada, M. Noda, S. Obika, Ca2+ enrichment in culture medium potentiates effect of oligonucleotides. Nuleic Acids Res., 2015, 43, e128. 7) K. Nishina et al., DNA/RNA heteroduplex oligonucleotide for highly efficient gene silencing. Nat. Commun., 2015, 6, 7969. 8) T. Yamaguchi, M. Horiba, S. Obika, Synthesis and properties of 2’-O,4’-C-spirocyclopropylene bridged nucleic acid (scpBNA), an analogue of 2’,4’-BNA/LNA bearing a cyclopropane ring. Chem. Commun., 2015, 51, 9737-9740. 9) A. R. Shrestha, Y. Kotobuki, Y. Hari, S. Obika, Guanidine bridged nucleic acid (GuNA): an effect of cationic bridged nucleic acid on dna binding affinity. Chem. Commun., 2014, 50, 575-577. 7 Laboratory of Molecular and Cellular Physiology Professor Kazutake TSUJIKAWA +81-6-6879-8190 [email protected] Specially Appointed Associate Professor So-ichiro FUKADA +81-6-6879-8191 [email protected] Specially Appointed Assistant Professor Hiroaki HASE +81-6-6879-8192 [email protected] Specially Appointed Assistant Professor Yuko UEDA +81-6-6879-8192 [email protected] Specially Appointed Reseacher Kaori KITAE +81-6-6879-8192 [email protected] FAX +81-6-6879-8194 Professor Kazutake TSUJIKAWA The 60 trillion cells that make up our bodies perform specialized functions according to genetic information encoded in DNA. Because these cells communicate and coordinate their functions, we are able to perform intelligent and active biological activity. However, functional disturbance of molecules and cells causes various kinds of diseases, including cancer and immune diseases, etc. In the Laboratory of Molecular and Cellular Physiology, we are researching the mechanisms of these biological activities and are developing the following projects to address cancer, RNA epigenetics, molecular-targeted drugs, and nerve-immune crosstalk. Cancer is a leading cause of death in our country. To better understand the molecular mechanism of carcinogenesis, we are identi- fying genes that are differentially expressed in normal and cancerous tissues, and are examining the function of genes. Prostate cancer antigen-1 (PCA-1) is one of the molecules we identified. PCA-1 is highly expressed in prostate cancer, non–small-cell lung cancer, and pancreatic cancer. Highly PCA-1 expression is significantly linked to poor prognosis in cancer patients. These findings indicate that PCA-1 would be a potential therapeutic target for the cancers. We are developing a molecular-targeted drug for PCA-1. Over 100 structurally distinct RNA modifications have been identified so far. These post-transcriptional modifications are widely present in various RNAs, including tRNA, mRNA, rRNA, miRNA, long non-coding RNA, etc. The post-transcriptional modifica- tion is called RNA epigenetics. However, the enzymatic regulatory mechanisms as well as the biological significance of RNA modification are not yet fully understood and are attracting the most attention in current RNA research. We are examining relation- ship among RNA epigenetics, cellular functions, and cancer development and progression. E. coli AlkB protein is a 2-oxoglutarate- and Fe(II)-dependent oxygenase that catalyzes the direct oxidative demethylation of methylated DNA and RNA bases. In humans, the AlkB homolog family consists of nine members (ALKBH1–ALKBH9), and PCA-1 is now called ALKBH3. Some of the ALKBH family molecules were found to highly express in cancer cells. Moreover, we identified high expression of some molecules including miRNAs in cancer. These molecules also play important functions in cancer cells. Our goal is to develop a potent, first-in-class molecular-targeted cancer drug. The immune and nervous systems are cross-talking via soluble mediators and their receptors. Recently, we reported that a neuro- peptide CGRP regulates functions of T cells and antigen-presenting cells. The immunomodulatory and tumor regulation mecha- nisms of CGRP are examining by using gene knockout mice. Research topics 1) Molecular mechanisms of RNA epigenetics 2) Molecular and cellular physiology of the ALKBH family molecules 3) Investigation and application of novel target molecules for cancer 4) Analysis of the nerve-immune-tumor axis 5) Identification and functional regulation of stem cells Recent publications 1) Ooshio I. et al., ALKBH8 promotes bladder cancer growth and progression through regulating the expression of surviving. Biochem. Biophys. Res. Commun. in press 2) Egawa H. et al., The miR-130 family promotes cell migration and invasion in bladder cancer through FAK and Akt phosphory- lation by regulating PTEN. Sci Rep. 6, 20574, 2016 3) Yamaguchi M. et al., Calcitonin Receptor Signaling Inhibits Muscle Stem Cells from Escaping the Quiescent State and the Niche. Cell Rep. 13, 302-14, 2015 4) Jingushi K. et al., miRNA-629 Targets TRIM33 to Promote TGF-beta/Smad Signaling and Metastatic Phenotypes in ccRCC. Mol. Cancer Res. 13, 565-74, 2015 8 Laboratory of Toxicology and Safety Science Professor Yasuo TSUTSUMI +81-6-6879-8230 [email protected] Associate Professor Kazuya NAGANO +81-6-6879-8231 [email protected] Assistant Professor Kazuma HIGASHISAKA +81-6-6879-8233 [email protected] FAX +81-6-6879-8234 HP http://www.phs.osaka-u.ac.jp/homepage/b009/ Visiting Professor Jian-Qing Gao (Zhejiang University) rosseforP Visiting Associate Professor Hiroko Shibata (NIHS) Visiting Teacher Toshiki Sugita (PMDA) IMUSTUSTousaY Visiting Researcher Shunji Imai, Tomohiro Nakao Toxicology/Safety Science is a branch of the pharmaceutical sciences dealing with the safety of pharmaceutical products derived from organic compounds, proteins, and genes as well as chemicals (xenobiotics) contained in cosmetics, food additives, and environmental pollutants. The objectives of Toxicology/Safety Science are to improve the human health and welfare, and to establish a secure and a safe society, including environmental protection. Toxicology/Safety Science, thus, contributes to human health and environmental protection through i) the underlying toxic mechanisms of adverse effects caused by medical products and various chemicals including biologics (antibody, cytokine, or vaccine) and nanomedicine, ii) providing assistance for ensuring the security by risk communication and risk literacy as well as the development of safer drugs, cosmetics, and foods, iii) evaluation of the influence of medicinal and chemical products on human health and the environment, and ensuring safety if deliberate and accidental exposure to those products occurs, and iv) establish- ment of effective coping strategies, methods for detoxification and treatment of drug-induced toxicity and environmental pollution. Based on these situations, our laboratory focused on the development of safe as well as useful innovative drug and risk analysis of nanomaterials (exposure assessment/analysis, hazad identification/analysis). In the course of these studies, we are attempting to elucidate both the mechanisms underlying toxicity caused by various environmental chemicals, and the physiological defense mechanisms that ordinarily protect living organisms against toxicity, using molecular cell biology, pharmacokinetics/pharmacodinamics and omics technol- ogy, and protein engineering to develop methods for detoxification. The following is an example of research on safety analysis of nanoma- terials (Nano-Safety Science Research). Nanomaterials have a variety of useful functions based on their increased surface area, electron reactivity, photo-reactivity, and thermal reactivity, as well as infrared emission because of their nanoscale size. Thus, these versatile materials are expected to have a wide range of beneficial properties. However, nanomaterials have pharmacokinetics and environmental kinetics that differ from those of existing micromaterials. As a result, nanomaterials may pose unpredictable biological effects and can act as a double-edged sword. Much of the current nanotechnology research focuses on practical applications for example, a number of cosmetic products on the market contain nanomaterials. Deliberate and accidental exposure to those products is already unavoidable, and such exposures could affect the fate of a number Figure 1. Clarification of the three dimensional conformation of tumor necrosis alpha receptor of related industries. Therefore, Nano-Safety Science Research, assistance of 2, which is one of the key molecule of immunotoxicity (Sci. Signal., 2010) safe forms of nanomaterial development as well as understanding and minimizing these unpredictable biological effects, is an urgent priority. The identification of safety biomarkers able to predict adverse events is an impor- tant initial step in the commercial development nanomaterials. Towards that goal, we are evaluating various characteristics of nanomaterials, including immunotoxicity, reproductive/developmental toxicity, genetic toxicity (mutagenicity), and oncogenicity. Furthermore, we are searching for safety/risk biomarkers (e.g., proteins, genes), establishing techniques for the prediction and evaluation of safety, and assisting the development of safe forms of nanomaterials by fusing toxico-kinetics analysis using imaging technologies, and toxico-proteomics. Using similar state-of-the-art technolo- gies, we are promoting studies to identify diagnostic biomarkers, drug targets and safety biomarkers for intractable diseases such as cardiac disorder, kidney disorder and cancers. Professor Hitoshi HASHIMOTO +81-6-6879-8180 [email protected] Specially Appointed Professor Toshio MATSUDA +81-6-6879-8161 [email protected] Associate Professor Norihito SHINTANI +81-6-6879-8181 [email protected] Assistant Professor Yukio AGO +81-6-6879-8168 [email protected] Assistant Professor Atsushi KASAI +81-6-6879-8182 [email protected] Assistant Professor (Molecular Research Center for Children's Mental Development) Professor Atsuko HAYATA +81-6-6879-8182 [email protected] Hitoshi HASHIMOTO The term,"a pharmacological action", means a response of living bodies to drugs.To know how the pharmacological actions are revealed, investigation of cellular and molecular mechanisms of living bodies is essential. In other words, pharmacology is, not only a scientific basis for the development of new drugs, but also largely contributes to the life science by providing many findings on the regulatory mechanisms of cells and organs. At present, when the total codes on the human genomes have being read, the functions of the human genomes becomes a main subject for life scientists to clarify. Thus, molecular pharmacology is an important academic field including gene technology (reverse pharmacology). 1.Genomics approaches todrugdiscovery For example, we are now showing the functions of new neuropeptides by using gene knockout mice which we recently generated. Neuroscience is a big science of the 21-century, because its progress has great. Impacts on a wide range of cultures including medicine and philosophy. In order to generate a novel drug, our lab is executing a systematic research for regulation of brain functions and responses to drugs in an aspect of molecular pharmacology. That is, our objects of the research extend from behavioral observations on whole animals to molecular biology on enzymes, drug receptors and genomes. The basis of brain functions is intracellular communications, which is regulated by a restricted recognition of transmitters and transduction of 2. Hyperactivity and abnormal jumping be havior of PACAP-knocko ut mice. Insets, examples of locomotor the signals between neurons in an appropriate manner. We are investigating the patterns during the last 10 min of a 60-min recording. signal transduction systems by means of molecular biological and neurochemical approaches and aim to generation of novel drugs. Some drugs examined in our lab are now applied to clinical trials. Research topics 1) Functional analysis of neuropeptide (PACAP) in brain and pancreas by molecular pharmacological approaches 2) Mechanisms for molecular basis of psychiatric function 3) Functional analysis of new biological target molecules for drug discovery 4) Molecular pharmacology of neural stem cell Recent publications 1) Nakamachi T, et al. PACAP suppresses dry eye signs by stimulating tear secretion. Nat Commun. 2016; 7: 12034. 2) Xu Z, et al. Pituitary adenylate cyclase-activating polypeptide (PACAP) contributes to the proliferation of hematopoietic progenitor cells in murine bone marrow via PACAP-specific receptor. Sci Rep. 2016; 6: 22373. 3) Nakazawa T, et al. Emerging roles of ARHGAP33 in intracellular trafficking of TrkB and pathophysiology of neuropsychiat- ric disorders. Nat Commun. 2016; 7: 10594. 4) Watanabe K, et al. Structured line illumination Raman microscopy. Nat Commun. 2015; 6: 10095. 5) Ohtaki H, et al. Pituitary adenylate cyclase-activating polypeptide (PACAP) decreases ischemic neuronal cell death in associa- tion with IL-6. Proc Natl Acad Sci USA. 2006; 103: 7488-7493. 6) Hashimoto H, et al. Altered psychomotor behaviors in mice lacking pituitary adenylate cyclase-activating polypeptide (PACAP). Proc Natl Acad Sci USA. 2001; 98: 13355-13360. 10 Laboratory of Biotechnology and Therapeutics Professor Shinsaku NAKAGAWA +81-6-6879-8175 [email protected] Associate Professor Naoki OKADA +81-6-6879-8176 [email protected] Assistant Professor Masashi TACHIBANA +81-6-6879-8177 [email protected] FAX +81-6-6879-8179 Professor Shinsaku NAKAGAWA The recent remarkable advance of the life science research brought a paradigm shift in "medicine". We can consider not only the conventional low molecular organic compounds but also bioactive proteins, nucleic acids, and cells to be as pharmaceutical materials. Our laboratory is studying themes based on "Drug Delivery System (DDS)" in fields of gene therapy, immunotherapy, and cell therapy. DDS on Vaccine Research (Transcutaneous vaccination system) The imminent practice of easy-to-use vaccination methods is expected to overcome some issues of injectable vaccinations. In addition, the development of new vaccination systems to enable worldwide mass treatment is critical for avoiding pandemics. An innovative approach that resolves these issues and is attracting great attention is the transcutaneous immunization (TCI) system. We are pushing forward with DDS research concerning the optimization and the practical use of TCI formulation. DDS on Cell Therapy Research (CAR-T cell therapy) The chimeric antigen receptor (CAR)-T cell therapy is moving toward the realization of its clinical application. We focused on vascular endothelial growth factor receptor 2 (VEGFR2) as a highly desirable target molecule for CAR-T cell therapy, because VEGFR2 abundantly exists on endothelial cells of tumor blood vessels whereas normal blood vessels express few VEGFR2. Now, we are planning clinical research of this promising novel CAR-T cell therapy for the verification of the safety and efficacy in human. In addition, we are attempting to acquire fundamental information of the CAR structure-activity correlation by the comparison analysis of various CAR derivatives. ''6RQ9DFFLQH5HVHDUFK ''6RQ&HOO7KHUDS\5HVHDUFK QIHFWLRXVGLVHDVHV 9DFFLQH ؼ 3URSK\ODFWLF PHGLFLQH WDUJHWLQJ LPPXQH V\VWHP, AlotofLQMHFWDEOH YDFFLQH is put to practical use, ற $,'6 ற 崎崏崡崰崲崌嵓 but approximately 4,000 people per day die of preventable ற 7KHPRVWFRPPRQFDXVHRIGHDWK$,'6 6$56 infectious diseases with vaccine +1 3DWLHQW ೭崌嵛崽嵓崐嵛崞 嵑崫崝 嵆嵑嵒崊 $,'6 嵆嵑嵒崊 &ROOHFWLRQRI ([SDQVLRQ &HOOXODUPHGLFLQH 7KHFULVLVRISDQGHPLFRILQIHFWLRQV嵆嵑嵒崊 +1 3DLQ DQG QHHGOHUHODWHG GLVHDVHV RU ᆈ崌嵛崽嵓崐嵛崞 SURJHQLWRUFHOOV 'LIIHUHQWLDWLRQ &\WRPHGLFLQH 崐嵄嵑লഷ LQMXULHV 7KHWKUHDWRIELRWHUURULVPDWWDFN崳崹崎崌嵓崡ഉඪ 嵆嵤嵓崾嵓崘লഷ 崷嵛崧崎崌嵓崡ཡඪණ 5HTXLUHPHQW IRU WUDLQHG SHUVRQQHO ,PPXQHFHOOWKHUDS\ 6WRUDJH RU WUDQVSRUW LVVXHV VXFK DV 'HQGULWLFFHOOV 7KHPRVWHIIHFWLYHFRXQWHUPHDVXUH FROG FKDLQ 7FHOOV LVSUHYHQWLRQWKURXJKYDFFLQDWLRQ 5HJHQHUDWLYHWKHUDS\ 3RRU YDFFLQH FRYHUDJH LQ GHYHORSLQJ FRXQWULHV 6WHPFHOOV ŦƈƏƏŃŧƈƏƌƙƈƕƜŃŶƜƖƗƈƐ 'HYHORSPHQWRIHDV\WRXVHDQGORZLQYDVLYHYDFFLQHSODWIRUP 7UDQVIHU 6HOIGLVVROYLQJ +\GURJHOSDWFK PLFURQHHGOHSDWFK 7UDQVFXWDQHRXVYDFFLQDWLRQV\VWHP 1H[WJHQHUDWLRQ&\WRPHGLFLQH *HQHSURWHLQ 'HVLJQDQGFUHDWLRQRIWKHRSWLPL]HGFHOOXODUIRUPXODWLRQ WUDQVGXFWLRQ WHFKQRORJ\ 7KH ZD\ RI XVLQJ LV VLPSOH DQG HDV\ WR MXVW SXW LW IRULQGLYLGXDOWUHDWPHQWVWUDWHJLHV ƈőƊő 0HGLFDO ZRUNHUV DUH XQQHFHVVDU\ IRU YDFFLQDWLRQ 7DUJHWLQJPROHFXOHH[SUHVVLRQ 7UDQVSRUWDWLRQ DQG VWRUDJH PD\ EH SRVVLEOH LQ 57 o $XJPHQWDWLRQRIORFDODFFXPXODWLRQ $SRSWRVLVLQKLELWRU\IDFWRUH[SUHVVLRQ 0HGLFDO ZDVWH ZLWK WKH YDFFLQDWLRQ GRHV QRW DSSHDU o ,PSURYHPHQWRIƌƑŃƙƌƙƒ SHUVLVWHQFH 9LUDOYHFWRU /LSRVRPH &KLPHULFDQWLJHQUHFHSWRUH[SUHVVLRQ o $GGLWLRQRIDQWLJHQVSHFLILFLQMXU\ $JUHDWGHPDQGIRUSUDFWLFDOWUDQVFXWDQHRXVYDFFLQHIRUPXODWLRQLV H[SHFWHGWKURXJKRXWWKHZRUOGLQFOXGLQJGHYHORSLQJFRXQWULHV Research topics 1) Development of transcutaneous vaccination/immunotherapy systems 2) Development of “Cell Delivery System” for the optimization of cell therapy 3) Investigation of the mechanisms of immune suppression for tumor immunotherapy Recent publications 1) H. Kusabuka, et al.: Biochem. Biophys. Res. Commun. 473(1): 73-79: (2016) 2) S. Hirobe, et al.: Biomaterials 57: 50-58 (2015) 3) K. Matsuo, et al.: J. Neuroimmunol. 266(1-2): 1-11 (2014) 4) S. Hirobe, et al.: Pharm. Res. 30(10): 2664-2674 (2013) 5) Y. Hiraishi, et al.: J. Control. Release 171(2): 93-103 (2013) 6) M. Yoshikawa, et al.: Blood 121(14): 2804-2813 (2013) 7) N. Kanagawa, et al.: Cancer Gene Ther. 20(1): 57-64 (2013) 11 Laboratory of Natural Products Chemistry Professor Motomasa KOBAYASHI +81-6-6879-8215 [email protected] Associate Professor Masayoshi ARAI +81-6-6879-8217 [email protected] Assistant Professor Naoyuki KOTOKU +81-6-6879-8216 [email protected] FAX +81-6-6879-8219 Professor Motomasa KOBAYASHI Bioassay-guided Total Synthesis separation Bioassays Medicinal Plants SAR study Bioactive Development of Marine Sponges Drug Lead Molecular-targeted anticancer drugs Substances Drug Design 1. Anti-angiogenic substance Marine Organisms Structure 2. Anti-metastasis substance elucidation Mechanistic Study 3. Cell-cycle inhibitor 4. Hypoxia-selective growth inhibitor Br Total synthesis O Antibiotics O furospinosulin-1 1. Anti-dormant mycobacterial substance PhO S 2. Inhibitor of biofilm formation N Hypoxia-selective growth inhibitor 2 H 3 Mechanistic analysis HO antitumor H evaluation Drugs for neurodegenerative disease O 1. Neuroprotective agent HO cortistatin A from oxidative stress N Anti-angiogenic substance Design and Asymmetric Synthesis of Total Synthesis Drug Leads Mechanistic Study by Chemical Genomics OMOM H N H N Target ID O H Mechanistic analysis TBSO I Genomic DNA Library Transformants O OMOM halicyclamine A Compound-resistant Anti-dormant Strains mycobacterial substance Medicinal plants and microorganisms (e.g., actinomyces) have been a rich source for new drugs for the last century. So far, ca. 25% of pharmaceuticals have been developed from natural products, which were isolated mainly from medicinal plants. These bioactive constituents may offer interesting subjects of study from the viewpoint of bioorganic chemistry as well as chemical biology and pharmaceutical chemistry. In search of new seeds, which have potential to be developed into new medicines, we have been studying bioactive marine natural products isolated from marine sponges and marine microorganisms together with medici- nal plants. We have been engaged in chemical studies on the following subjects. Research topics 1) Search for new bioactive natural products guided by constructed new bioassay systems. 2) Structural elucidation of bioactive substance on the basis of spectral analysis and synthetic method. 3) Asymmetric total synthesis of bioactive substance. 4) Synthetic structure-activity relationship study of bioactive substance to elucidate pharmacophores. 5) Mechanistic study of bioactive substance by techniques of chemical biology and chemical genetics. Recent publications 1) Arai, M. et al. Furospinosulin-1, marine spongean furanosesterterpene, suppresses the growth of hypoxia-adapted cancer cells by binding to transcriptional regulators p54nrb and LEDGF/p75. ChemBioChem, 17, 181-189 (2016) 2) Arai, M. et al. Identification of target protein of agelasine D, a marine spongean diterpene alkaloid, as an anti-dormant myco- bacterial substance. ChemBioChem, 15, 117-123 (2014) 3) Kotoku, N. et al. Creation of Readily Accessible and Orally Active Analogue of Cortistatin A. ACS Med. Chem. Lett., 3, 673-677 (2012) 4) Kotoku, N. et al. Stereoselective Synthesis of Core Structure of Cortistatin A. Org. Lett., 13, 3514-3517 (2011) 5) Aoki, S. et al. Cortistatins A, B, C, and D, Anti-angiogenic Steroidal Alkaloids, from the Marine Sponge Corticium simplex. J. Am. Chem. Soc., 128, 3148-3149 (2006) 12 Laboratory of Clinical Science and Biomedicine Professor Yasushi FUJIO +81-6-6879-8258 [email protected] Associate Professor Hiroyuki NAKAYAMA +81-6-6879-8252 [email protected] Assistant Professor Masanori OBANA +81-6-6879-8163 [email protected] FAX +81-6-6879-8253 Professor Yasushi FUJIO One of the most important goals of life science is to contribute One of the most significant unanswered questions in cardiac to the happiness of human beings through its clinical applica- biology is how calcium-dependent signaling pathways are tion, but this does not mean that the clinical application is the regulated against the backdrop of the Ca2+ transient that medi- final step of life science. Clinical findings often propose ates contraction. To address this question, we assessed the important questions to basic scientists and encourage them to roles of multiple calcium-related proteins and finally found develop novel scientific fields. We recognize the limitations that enhanced Ca2+ influxes via L-type Ca2+ channels (LTCC) and disadvantages of up-to-date pharmacotherapies, mainly in mediated cardiac cell death and heart failure after cardiovascular diseases, and address these problems by the ȕ‐adrenergic stimulation which also plays a central role in integrated approaches, including molecular and cellular developing heart failure. Currently, we are scrutinizing the biology and pharmacogenomics. Three projects are on-going. patho- physiological roles of regulatory subunits of LTCC in [Project 1] Cytokine science in cardiovascular diseases ȕ‐adrenergic signaling in heart failure as follows. 1) Phos- Cardiac functions are beneficially or detrimentally modulated phorylation of the regulatory subunit of LTCC; The regulatory by various kinds of cytokines. In this project, we will make subunit of LTCC is phosphorylated and subsequently activated clear the cytokine network in diseased hearts and establish by two protein kinases, cAMP- dependent protein kinase and novel therapies that target cytokine signals. We address this Ca2+/calmodulin-dependent protein kinase both of which are issue from two viewpoints. 1) Cardioprotective cytokines: We DFWLYDWHG GRZQVWUHDP ȕ‐adrenergic receptor signaling. We have been investigating the pathophysiological roles of IL-6 are scrutinizing the role of those phosphorylation using geneti- family cytokines in cardiovascular diseases and found that cally engineered mice. 2) Mechanism of cell death during some cytokines, such as leukemia inhibitory factor and IL-11, enhanced Ca2+ influxes. The hearts with enhanced Ca2+ are cardioprotective and preserve myocardium through the influxes demonstrated a histologically unique form of cell activation of STAT3, a signal transducer molecule. Now we death. We will elucidate the molecular mechanisms of this are planning a clinical trial to examine whether IL-11 treatment orphan cell death utilizing genetically engineered mouse protects myocardium in acute myocardial infarction. 2) models. Inflammation and heart failure: Heart failure is closely associ- [Project 3] Discovery of novel therapeutic targets and ated with inflammation. Serum concentrations of proinflam- biomarkers for chronic kidney disease (CKD). CKD is a PDWRU\F\WRNLQHVLQFOXGLQJ,/DQG71)ĮLQFUHDVHLQWKH worldwide public health problem because it is not only a patients with heart failure; however, it remains to be elucidated leading cause of end-stage renal disease but also is a major risk how inflammatory reactions sustain in the failing hearts. We factor for cardiovascular disease. However, the pathogenesis are investigating the molecular and cellular mechanisms of of CKD remains to be elucidated. Moreover, there are few chronic inflammation in heart failure. medicine for CKD. In this project, we investigate novel thera- [Project 2] Calcium-mediated pathogenesis in heart peutic targets and biomarkers for CKD using various animal failure. Calcium ion (Ca2+) plays pivotal roles in excitation- disease models and samples from patients. The project is contraction coupling and activation of calcium-dependent expected to make a great contribution to medical development reactive signaling both of which underpin the development of in kidney disease. heart failure. The concentrations of Ca2+ are distinct in various organelle and well-regulated by a great number of proteins. Recent publications 1) Miyawaki, A. et al. Moesin is activated in cardiomyocytes in experimental autoimmune myocarditis and mediates cytoskeletal reorganization with protrusion formation. Am J Physiol Heart Circ Physiol. In press. 2) Nakagawa, M. et al. Four cases of investigational therapy with interleukin-11 against acute myocardial infarction. Heart and Vessels. In press. 3) Kumagai, S. et al. Myeloid cell-derived LRG attenuates adverse cardiac remodeling after myocardial infarction. Cardiovasc. Res. 109, 272-282. 2016. 4) Enomoto, D. et al. Cardiac specific ablation of STAT3 gene in subacute phase of myocardial infarction exacerbated cardiac remodeling. Am. J. Physiol. Heart Circ. Physiol. 309, H471-480. 2015. 5) Matsuo, R.et al. The inhibition of N-glycosylation of glycoprotein 130 molecule abolishes STAT3 activation by IL-6 family cytokines in cultured cardiac myocytes. PLoS One 9, e111097. 2014. 6) Okamoto, H. et al. Minimal dose for effective clinical outcome and predictive factors for responsiveness to carvedilol: Japa- nese chronic heart failure (J-CHF) study. Int. J. Cardiol. 164, 238-244. 2013. 7) Obana, M. et al. Therapeutic administration of IL-11 exhibits the postconditioning effects against ischemia-reperfusion injury via STAT3 in the heart. Am. J. Physiol. Heart Circ. Physiol. 303, H567-H577. 2012. Professor Fumihiro OKADA +81-6-6105-5792 [email protected] Associate Professor Fumitaka FUJITA +81-6-6105-5792 [email protected] Visiting Academic Staff Ryuichiro KURATA +81-6-6105-5792 [email protected] FAX +81-6-6879-8154 Professor Tadayuki UNO +81-6-6879-8205 [email protected] Professor Yasushi FUJIO +81-6-6879-8258 [email protected] Advanced Medical Technologies The technologies of regenerative medicine, such as culture of stem-cells and regenerating organs, are dramatically developed. Many people believe that these technologies should be applied to skin regeneration and regulation for physiological function of cells, resulting development of cosmetics. We collaborate with medical schools and clinics to utilize 㻴㼡㼙㼍㼚 㻵㼟㼛㼘㼍㼠㼕㼛㼚㻌㼛㼒 㻿㼜㼔㼑㼞㼑㻌㼒㼞㼛㼙 human isolated skin by surgical operations. We analyze protein 㻿㼣㼑㼍㼠㻌㼓㼘㼍㼚㼐 㼟㼠㼑㼙㻌㼏㼑㼘㼘 㼟㼕㼚㼓㼘㼑㻌㼟㼠㼑㼙㻌㼏㼑㼘㼘 and gene expression or isolation of stem-cells for analysis of cell function and differentiation using FACS and confocal micros- copy, etc. Revolutionary cosmetics For comfortable life of all people, advanced medical technolo- 㻴㼡㼙㼍㼚㻌㼟㼗㼕㼚 㻯㼑㼘㼘㻌㼎㼍㼚㼗 gies including regenerative medicine are applied to research skin 㼎㼥㻌㼟㼡㼞㼓㼕㼏㼍㼘㻌㼛㼜㼑㼞㼍㼠㼕㼛㼚 and other organ homeostasis. We have tried to develop the revolutionary cosmetics. Since so many people have skin trouble 㼀㼕㼟㼟㼡㼑 that we have to realize novel technologies for the effective 㻻㼞㼓㼍㼚㼟 㻯㼑㼘㼘㼟 products from our basic research finding. All of our research themes are designed to get novel findings from the bench to the consumers speedily, called “Bench to Consumer”. Bench to Consumer 㻼㼞㼛㼠㼑㼕㼚㻌㻒㻌㻳㼑㼚㼑 㻯㼑㼘㼘㻌㼒㼡㼚㼏㼠㼕㼛㼚 So far, we had developed novel cosmetic technologies from basic science. “Antimicrobial agents-free” was based on finding the anti-microbial function of “alkandiol”. “Deodorant for middleage” was based on discovery the odor chemicals “diace- til”. Through study about sensor on skin surface, “TRP channels”, “eucalyptol” was found as a comfortable cooling agent. Research topics 1) Application of advanced medical technology for revolutionary cosmetics 2) Maintenance for interfollicular stem cells for developing skin drug 3) Dynamic shrinking under the molecular basis of regulation for sweat gland 4) Cross-talking between melanoblast and hair follicle stem cells for coloring or de-coloring of human hair 5) Molecular mechanisms for the detection of extracellular changes of immune cells in skin Recent publications 1) Takaishi M. et al., Reciprocal effects of capsaicin and menthol on thermosensation through regulated activities of TRPV1 and TRPM8. J Physiol Sci. 66, 143-155, 2016 2) Kurata R. et al., Isolation and characterization of sweat gland myoepithelial cells from human skin. Cell Struct. Funct. 39, 101-112, 2014 3) Fujita F. et al., Ambient temperature affects the temperature threshold for TRPM8 activation through interaction of phosphati- dylinositol 4,5-bisphosphate. J. Neurosci. 33, 6154-6159, 2013 4) Takaishi M, et al., 1,8-cineole, a TRPM8 agonist, is a novel natural antagonist of human TRPA1. Mol. Pain. 8:86, 2012 14 Laboratory of Molecular Medicine Professor Takefumi DOI +81-6-6879-8158 [email protected] Associate Professor Yoshiaki OKADA +81-6-6879-8164 [email protected] Assistant Professor Nobumasa HINO +81-6-6879-8165 [email protected] FAX +81-6-6879-8158, +81-6-6879-8164 Professor Takefumi DOI The information in the genome is transcribed to RNA and then 1RQ(& translated to protein. Protein plays various and critical roles in living cells. (6FHOO 2)) For optimal protein functions, protein molecules need to form proper 2)) three dimensional structures with optimal post-translational modifications SUR[LPDOSURPRWHU and interact with their target molecules. It is also necessary that optimal amount of protein is expressed in appropriate tissue in appropriate timing. These regulations are essential for the physiological protein function and homeostasis in health. However, once the protein structure, modification, (& and spatial and temporal regulation of protein expression are disrupted, 21 disease is induced. It has been demonstrated that aberrant protein expression caused by gene defects, such as mutation and translocation, ectopic gene expression and abnormal protein expression level are involved in the Endothelialcell(EC)ͲspecificRobo4expressionregulated induction of diseases. But, there are still many diseases to be investigated. bytranscriptionfactorsandDNAmethylation One of the goals of our lab is to reveal the molecular mechanism for those diseases, and propose the novel strategy to cure them. To this end, we are focusing on the proteins which are closely related to disease and investigating their physiological functions in health and disease. We are now investigating transcription factors and epigenetic control related to cell type-specific gene expression in endothelial cells. We also try to understand functions of the endothelial cell-specific receptor and nuclear receptors through identification of their binding partners by unique in vivo photo-crosslinking technique. From these information we would like to propose novel mechanisms for inflammatory diseases and metabolic syndrome. Research topics 1) Functions and transcriptional regulation of endothelial cell-specific receptor, Robo4 2) Generation of new drug candidates against inflammatory diseases by targeting Robo4 3) In vivo protein photo-crosslinking with genetically encoded non-natural amino acids 4) Structure and function of nuclear receptors and histone methyltransferase Recent publications 1) Okada Y. et al., Expression of the Robo4 receptor in endothelial cells is regulated by two AP-1 protein complexes. Biochem. Biophys. Res. Commun., 467(4), 987-991, 2015 2) Matsumura Y. et al., H3K4/H3K9me3 Bivalent Chromatin Domains Targeted by Lineage-Specific DNA Methylation Pauses Adipocyte Differentiation. Mol. Cell., 60(4), 584-596, 2015 3) Tachibana K. et al., Analysis of the subcellular localization of the human histone methyltransferase SETDB1. Biochem. Biophys. Res. Commun., 456, 725-731, 2015 4) Shimo T. et al., Design and evaluation of locked nucleic acid-based splice-switching oligonucleotides in vitro. Nucleic Acids Res., 42(12), 8174-8187, 2014 7DQDND7HWDO33$5ȕįDFWLYDWLRQRI&'DFRQWUROVLQWHVWLQDOLPPXQLW\Sci. Rep., 4:5412, 1-9, 2014 6) Okada Y., et al., Endothelial cell-specific expression of Robo4 is regulated by differential DNA methylation of the proximal promoter. Arterioscler. Thromb. Vasc. Biol., 34 (7), 1531-1538, 2014 7) Yamaguchi S. et al., Selective evaluation of high density lipoprotein from mouse small intestine by an in situ perfusion technique. J Lipid Res., 55, 905-918, 2014 8) Okada Y. et al., RUNX1, but not its familial platelet disorder mutants, synergistically activates PF4 gene expression in combi- nation with ETS family proteins. J.Thromb. Haemost., 11, 1742-1750, 2013 9) Tachibana K. et al., Human mannose-binding lectin 2 is directly regulated by peroxisome proliferator-activated receptors via a peroxisome proliferator responsive element. J. Biochem., 154, 265-273, 2013 10) Yoshikawa M. et al., Robo4 is an effective tumor endothelial marker for antibody-drug conjugates based on the rapid isolation of the anti-Robo4 cell-internalizing antibody. Blood, 121, 2804-2813, 2013 11) Yakura Y. et al., An induced pluripotent stem cell-mediated and integration-free factor VIII expression system. Biochem. Biophys. Res. Commun., 431, 336-341, 2013 15 Department of Preemptive Medical Pharmacology for Mind and Body Endowed Chair Professor Atsuo SEKIYAMA +81-6-6879-4093 [email protected] Endowed Chair Assistant Professor Emiko KASAHARA +81-6-6879-4094 [email protected] FAX +81-6-6879-4093 Professor Atsuo SEKIYAMA Our Aim Our aim is to provide a preventative way against diseases. We focus ourselves not only to therapy of diseases, but also develop- ment of diagnostics, which give us chances of prevention of an onset, exacerbation, and recurrence of diseases. Ongoing Investigations 1) Objectifying the pathway by which psychological stress induces diseases. It has been suggested that there underlie a relationship between onset and exacerbation of diseases and psychological stress, while the pathway nor mechanism is understood. One of our missions is to elucidate a molecular pathway by which psychological stress enhances vulnerability of host-defense, and one of our achievements is clarification of a psychological stress induced, super-oxide mediated caspase-1 activation pathway in the adrenal gland that causes up-regulation of interleukin (IL) -18 in circulating blood, resulting in an altered regulation of immunity and inflammation of a host. Another of ours was demonstration of cytokines releas- ing pathway that is induced by mitochondrial stress. Thus, we carry our investigations with pleural eye-sites, one is systemic level of whole body and the other is ‘micro’, influencing to our research style and opinion on all living things. 2) Development of diagnostic pharmaceutical for risk of ‘onset’ of depression. Depression after psychological stress has been suggested to be one of the avoidable diseases, however, there has been no measure to indicate us the time for preventative intervention to its potent victims. We have found that a profile of cytokines levels in plasma are altered after stress and that the levels of the alteration is associated with risk of depression after stress. Basing on these findings, the detection of high-risk individuals for depression after stress has been employed for people exposed to stressful event (e.g. Tohoku earthquake), and, on the other hand, project for providing an indicating pharmaceutical for a risk of an onset of depression has been performed. 3) Development of diagnostic pharmaceutical for psychiatric diseases There has been no measure to objectify depression and schizophrenia but an interview by specialists such as psychiatrist and psychologist, whose number is much less than sufferers by those diseases, resulting in a loss of opportunities of assessment for appropriate treatment. We have found that a profile of cytokines levels in plasma are perturbed in those diseases and can be used as indicators for depression and schizophrenia. Basing on these findings, development of indicating pharmaceutical for depression and schizophrenia has been performed. A goal of the project is establishment of indicators of depression and schizophrenia for clinical scenes. Research topics 1) Investigation for a pathway by which psychological stress induces diseases. 2) Development of diagnostic pharmaceutical for risk of ‘onset’ of depression. 3) Development of diagnostic pharmaceutical for depression. 4) Development of diagnostic pharmaceutical for schizophrenia. 5) Development of therapeutic drugs for schizophrenia. Recent publications 1) Physiological stress exacerbates murine colitis by enhancing pro-inflammatory cytokine expression that is dependent. Am J Physiol Gastrointest Liver Physiol. 2011, 301(3):G555-64. 2) Mitochondrial density contributes to the immune response of macrophages to lipopolysaccharide via the MAPK pathway. FEBS Lett. 2011, 585(14):2263-8. 3) A stress-induced, superoxide-mediated caspase-1 activation pathway causes plasma IL-18 upregulation. Immunity. 2005, 22(6):669-77. 4) International patent application: Biological load indicator and method of measuring biological load. PCT/JP2007/052887 (Patented in Japan, No.5069213) 16 Laboratory of Pharmainformatics and Pharmacometrics Professor Tatsuya TAKAGI +81-6-6879-8243 [email protected] Assistant Professor Norihito KAWASHITA +81-6-6879-8243 [email protected] FAX +81-6-6879-8243 Professor Tatsuya TAKAGI Our group is interested in Pharmacometrics including pharmacoinformatics, clinical pharmacometrics, environmental pharmacometrics, and social pharmacoinformatics. Pharmacometrics is considered to be a new term in the field of pharmaceutical sciences. In this century, advanced information society has been already realized. Any kind of information immediately is able to be accessed by cellular phones. Moreover, researchers have been utilizing powerful computing systems such as mobile computers. Pharmacometrics is considered as one of the most important fields which will be able to play an important role in big data analysis. “Social, Clinical, and Environmental Pharmacometrics” In the case of pharmacoepidemiological studies, the linear method which called generalized linear model is usually adopted. However, the relationships between risk factors and link functions of clinical data are not always linear. Recently, nonparametric regression analysis attracts considerable interest as a solution methodology in such cases. It is unnecessary to specify the function which represents the relationship between predictor and response variables. Our group has been trying to develop new methods of nonparametric regression and discriminant analyses. “Computational Chemistry for infectious diseases and chemical reaction analysis” We study mechanisms of virulence and search a novel candidate compounds for some infectious diseases, such as HIV, influenza, and dengue virus by computational chemistry such as molecular dynamics and docking study. We also analyze mechanisms of organic reactions by molecular orbital calculation. Research topics 1) Quantitative Structure-Toxicity Relationship of environmental toxicity 2) In silico diagnostic imaging using machine learning 3) Development of novel regression analysis method 4) Protein-protein interaction analysis by quantum chemical calculation using supercomputer 5) Screening of anti-infection drugs by HTS and in silico method 6) Analysis of mechanism for side effect by molecular dynamics simulation and docking simulation 7) Informatics of thalidomide risk management 8) Analysis of chemical reaction by quantum chemical calculation Recent publications 1) Arai Y, Kawashita N, Daidoji T, Ibrahim MS, El-Gendy EM, Takagi T, Takahashi K, Suzuki Y, Ikuta K, Nakaya T, Shioda T, Watanabe Y. Novel Polymerase Gene Mutations for Human Adaptation in Clinical Isolates of Avian H5N1 Influenza Viruses. PLoS Pathog. 2016, 12, e1005583. 2) Okamoto Y, Tsuneto S, Morita T, Takagi T, Shimizu M, Miyashita M, Uejima E, Shima Y. Desirable Information of Opioids for Families of Patients With Terminal Cancer: The Bereaved Family Members' Experiences and Recommendations. Am J Hosp Palliat Care. in press. 3) Tian YS, Kawashita N, Arai Y, Okamoto K, Takagi T. Pharmacophore Modeling and Molecular Docking Studies of potential inhibitors to E6 PBM-PDZ from Human Papilloma Virus (HPV). Bioinformation. 2015, 11, 401-406. 4) Watanabe Y, Arai Y, Daidoji T, Kawashita N, Ibrahim MS, El-Gendy Eel-D, Hiramatsu H, Kubota-Koketsu R, Takagi T, Murata T, Takahashi K, Okuno Y, Nakaya T, Suzuki Y, Ikuta K. Characterization of H5N1 influenza virus variants with hemagglutinin mutations isolated from patients. MBio. 2015, 6, e00081-15. 5) Oka Y, Okamoto K, Kawashita N, Shirakuni Y, Takagi T. Meta-analysis of the risk of upper gastrointestinal hemorrhage with combination therapy of selective serotonin reuptake inhibitors and non-steroidal anti-inflammatory drugs. Biol Pharm Bull. 2014, 37, 947-953. 6) Pambudi S, Kawashita N, Phanthanawiboon S, Omokoko MD, Masrinoul P, Yamashita A, Limkittikul K, Yasunaga T, Takagi T, Ikuta K, Kurosu T. A small compound targeting the interaction between nonstructural proteins 2B and 3 inhibits dengue virus replication. Biochem Biophys Res Commun. 2013, 440, 393-398. 17 Laboratory of Applied Environmental Biology Professor Kazumasa HIRATA +81-6-6879-8236 [email protected] Lecturer Kazuo HARADA +81-6-6879-8235 [email protected] Assistant Professor Hideyuki MATSUURA +81-6-6879-8238 [email protected] Professor Kazumasa HIRATA As a result of rapid population growth and economic develop- &RQVWUXFWLRQRITXDOLW\ 3UHYHQWLRQRIRFFXUUHQFHRI FRQWUROPHWKRGVIRUPHGLFLQDO ment, we are facing with a wide variety of global health PXOWLGUXJUHVLVWDQFHEDFWHULD SODQWPDWHULDOV concerns including a food crisis, energy problem, environmen- 'HSOHWLRQRIPHGLFLQDO (PHUJLQJRUUHHPHUJLQJ SODQWPDWHULDOV tal pollution, or emerging/re-emerging infections disease. In LQIHFWLRXVGLVHDVHV (QYLURQPHQWDOUHVWRUDWLRQ this situation, roles of pharmaceutical sciences have been ,PSURYHPHQWRIIRRG XVLQJJHQHWLFDOO\PRGLILHGSODQWV SURGXFWLRQXVLQJJHQHWLFDOO\ +HDOWKFDUH expanding from a contribution to the development or proper use PRGLILHGFURSV of pharmaceuticals to the “environmental restoration and reme- (QYLURQPHQWDO GHJUDGDWLRQ diation”, “securement of food safety and security”, and “conser- )RRGVKRUWDJH )RRG (QYLURQPHQW vation of biodiversity and sustainable use of biological )RRGFRQWDPLQDWLRQ (QYLURQPHQWSROOXWLRQ resources”, i.e., expanding to comprehensive health promotion and maintenance. The aim of our laboratory is to contribute to (QHUJ\ &RQVWUXFWLRQRI (QYLURQPHQWDOUHPHGLDWLRQ the construction of sustainable society with securement of PDQDJHPHQWV\VWHPIRU (QHUJ\GHSOHWLRQ XVLQJIXQFWLRQVRI human health, environmental health, and food security/safety IRRGVHFXULW\DQGVDIHW\ SKRWRV\QWKHWLFRUJDQLVPV through the research topics listed below. %LRPDVVDSSOLFDWLRQVRI SKRWRV\QWKHWLFRUJDQLVPV Research topics 1) Analysis of stress response mechanisms to environmental stresses in photosynthetic organisms and its application to environmental restoration and remediation, food production, or production of pharmaceutical materials. 2) Development of quality control methods for medicinal plant materials using high tech analytical instruments. 3) Development of comprehensive monitoring system for environmental pollutants. Recent publications 1) Miyasaka et al., Is chloroplastic class IIA aldolase a marine enzyme? ISME Journal, in press. 2) Uchida et al., Monitoring of antibiotic residues in aquatic products in urban and rural areas of Vietnam. J. Agric. Food Chem., in press 3) Do et al., Screening of antibiotic residues in pork meat in Ho Chi Minh City, Vietnam, using a microbiological test kit and liquid chromatography/tandem mass spectrometry. Food Control, 69, 262-266 (2016) 4) Nguyen et al., Spread of antibiotic and antimicrobial susceptibility of ESBL-producing Escherichia coli isolated from wild and cultured fish in the Mekong Delta, Vietnam. Fish Pathology, 51, S75-S82 (2016) 5) Yamaguchi et al., Antibiotic residue monitoring results for Pork, Chicken, and Beef samples in Vietnam in 2012–2013. J. Agric. Food Chem., 63, 5141–5145 (2015) 6) Yamaguchi et al., Rapid and easy multiresidue method for the analysis of antibiotics in meats by ultrahigh-performance liquid chromatography–tandem mass spectrometry. J. Agric. Food Chem., 63, 5133–5140 (2015) 7) Matsushima et al., Immobilization of enzymatic extracts of Portulaca oleracea cv. roots for oxidizing aqueous bisphenol A. Biotechnol Lett., 37, 1037-1042 (2015) 8) Ogata et al., Functional characterization of a carotenoid cleavage dioxygenase 1 gene from Chlamydomonas reinhardtii. Carotenoid Science, 19, 28-32 (2014) 9) Matsuura et al., A computational and experimental approach reveals that the 5’-proximal region of the 5’-UTR has a cis-regulatory signature responsible for heat stress-regulated mRNA translation in Arabidopsis. Plant Cell Physiol., 54, 474-483 (2013) 10) Matsuura et al., Development of surface-engineered yeast cells displaying phytochelatin synthase and their application to cadmium biosensors by the combined use of pyrene-excimer fluorescence. Biotechnol Prog., 29, 1197-1202 (2013) 18 Laboratory of Clinical Pharmacy Research and Education Professor Etsuko UEJIMA +81-6-6879-8251 [email protected] Associate Professor Kazuyuki NIKI +81-6-6879-8250 [email protected] Specially Appointed Assistant Professor Asuka HATABU +81-6-6879-8250 [email protected] Professor Etsuko UEJIMA What patients require for Medicine? It may be said to be “High Quality and Effective Care.” It can be said that it consists of “Improvement of Therapeutic effect”, “Risk reduction”, and “Cost-effective”. In order to fulfill their requirements, “Team Approach to Health Care “is essential. We research collaborate with departments and agencies of Osaka University hospital and so on. It is said that there are five tools in clinical trial. New drugs are put to the test in postmarketing phase. So pharmacovigilance in early stage is especially important. There are many unsolved problems or questions in clinical practice. Our research heads for solving medical problems and resolv- ing clinical questions. prepared by E. Uejima Research topics 1) Clinical data base research 2) Clinical research on improving antibiotic prescribing for infection 3) Clinical research of palliative care 4) Clinical research on Cancer Chemotherapy 5) Surveillance, pharmacoepidemiology, and pharmacoeconomics Recent publications 1) Morio K, Minami T, Sozu T, Niki K, Kijima T, Uejima E：Weight Loss Associated with Platinum-Based Chemotherapy in Patients with Advanced Lung Cancer.: Chemotherapy.:61(5):256-61(2016) 2) Okamoto Y, Tsuneto S, Morita T, Takagi T, Shimizu M, Miyashita M, Uejima E, Shima Y: Desirable Information of Opioids for Families of Patients With Terminal Cancer: The Bereaved Family Members' Experiences and Recommendations. Am J Hosp Palliat Care: (2016) Jan 13. 3) Tsujimoto T, Yamamoto Y, Wasa M, Takenaka Y, Nakahara S, Takagi T, Tsugane M, Hayashi N, Maeda K, Inohara H, Uejima E, Ito T:L-glutamine decreases the severity of mucositis induced by chemoradiotherapy in patients with locally advanced head and neck cancer: a double-blind, randomized, placebo-controlled trial. Oncol Rep. :Jan;33(1):33-9. (2015) 19 Laboratory of Cell Biology Professor Kunihiko NISHINO +81-6-6879-8545 [email protected] Associate Professor Tsuyoshi NISHI +81-6-6879-8547 [email protected] Assistant Professor Mitsuko HAYASHI-NISHINO +81-6-6879-8545 [email protected] Assistant Professor Seiji YAMASAKI +81-6-6879-8546 [email protected] FAX +81-6-6879-8549 Professor Kunihiko NISHINO Xenobiotic transporters are widely distributed in living organisms and play important roles in their self-defense mechanisms. Transporters cause a serious chemotherapeutic problem named multidrug resistance in pathogenic bacteria and cancer cells. Recent discoveries also support the notion that some xenobiotic transporters have shown to have important roles in bacterial virulence and signal transduction. We are interested in understanding roles xenobiotic transporters in multidrug resistance and physiological functions. Our knowledge should promote the development of novel inhibitors or strategies that could counteract the contribution of xenobiotic transporters to drug resistance and virulence. Fig.1. Nanosystems biology of multidrug resistance. Fig.2. Mouse SPNS2 regulates lymphocyte egress into blood. Research topics 1) Development of therapeutic strategies to control infectious diseases 2) Regulation and physiological function of xenobiotic transporters 3) Nanosystems biology of multidrug resistance 4) Development of new devices to detect multidrug resistant bacteria 5) 3D imaging of bio-nanostructures by electron tomography Recent publications 1) Yamasaki S. et al. Phenotype microarray analysis of the drug efflux systems in Salmonella enterica serovar Typhimurium. J. Infect. Chemother., in press, 2016 2) Yamasaki S. et al. Multidrug efflux pumps contribute to Escherichia coli biofilm maintenance. Int. J. Antimicrob. Agents, 45, 439-441, 2015 3) Nishi T. et al. Molecular and physiological functions of sphingosine 1-phosphate transporters. Biochim. Biophys. Acta, 1841, 759-765, 2014 4) Horiyama T. & Nishino K. AcrB, AcrD, and MdtABC multidrug efflux systems are involved in enterobactin export in Escherichia coli. PLoS One, 9, e108642, 2014 5) Yamasaki S. et al. The crystal structure of multidrug-resistance regulator RamR with multiple drugs. Nature Commun., 4, 2078, 2013 6) Yamasaki S. et al. Cooperation of the multidrug efflux pump and lipopolysaccharides in the intrinsic antibiotic resistance of Salmonella enterica serovar Typhimurium. J. Antimicrob. Chemother., 68, 1066-1070, 2013 7) Nakashima R. et al. Structural basis for the inhibition of bacterial multidrug exporters. Nature, 500, 102-106, 2013 8) Hisano Y. et al. Mouse SPNS2 functions as a sphingosine-1-phosphate transporter in vascular endothelial cells. PLoS One, 7, e38941, 2012 9) Nakashima R. et al. Structures of the multidrug exporter AcrB reveal a proximal multisite drug-binding pocket. Nature, 480, 565-569, 2011 10) Kawahara A. et al. The sphingolipid transporter spns2 functions in migration of zebrafish myocardial precursors. Science, 323, 524-527, 2009 20 Laboratory of Biomolecular Science and Engineering Professor Takeharu NAGAI +81-6-6879-8480 [email protected] Associate Professor Tomoki MATSUDA +81-6-6879-8481 [email protected] Assistant Professor Yoshiyuki ARAI +81-6-6879-8481 [email protected] Assistant Professor Masahiro NAKANO +81-6-6879-8481 [email protected] Specially Appointed Assoc. Prof. Tetsuichi WAZAWA +81-6-6879-8481 [email protected] Specially Appointed Assist. Prof. Megumi IWANO +81-6-6879-8481 [email protected] Specially Appointed Assist. Prof. Tetsuyuki ENTANI +81-6-6879-8481 [email protected] Professor Takeharu NAGAI Comprehensive analysis of the spatio-temporal dynamics of individual biomolecules is key to uncovering the mechanism of physiological phenomena at molecular level, which leads to the application of drug discovery and tailor-made medicines. In our laboratory, we have been developing genetically encoded indicators for physiological functions by engineering fluorescent proteins and chemiluminescent proteins such as GFP and luciferase, respectively. Now we can observe gene expression, protein localization, protein diffusion, protein conformational change, protein-protein interaction, and dynamics of bioactive molecules such as ATP and Ca2+ not only in cell but also in vivo. In addition, we have been developing novel optical microscopies with companies as industry-university collaboration for more effective use of our visualization indicator toolbox. By combining these techniques, we are trying to elucidate the living system at nano meter resolution so as to promote the application of drug develop- ment and diagnosis devices. For example, development of an in vivo-based high-through-put screening system for lead compounds will change the way of drug development. Invention of an on-site measurement system that is based on the detection of signals from our chemiluminescent indicators by smartphones is among one of our current focus. This technology would enable quick inspection not at hospital but home, which contribute to the early detection of severe disease including tumors, as well as the improvement of health care system in developing countries that equip far insufficient medical institutions. 6XEVWUDWH DQDO\WH ,QYLYRGUXJHYDOXDWLRQDQGVFUHHQLQJE\ RXURULJLQDOFKHPLOXPLQHVFHQFHLQGLFDWRUV &RORUFKDQJHXSRQ OLJDQGELQGLQJ 6PDUWSKRQH 6DPSOHKROGHU /XPLQHVFHQFH :DYHOHQJWK Figure 1, Schematic diagram of the evaluation of molecules by Figure 2, Schematic diagram of on-site measurement system for using chemiluminescent probes in mouse. bioactive compounds by using chemiluminescent probes. Research topics 1) Development of sensors based on fluorescent and/or chemiluminescent proteins 2) Development of chemical compound screening system with using chemiluminescent biosensors 3) Development of on-site measurement for bioactive molecules Recent publications 1) Inagaki S. & Nagai T. Current progress in genetically encoded voltage indicators for neural activity recording. Curr. Opin. Chem. Biol. 33, 95-100, 2016. 2) Tiwari DK, et al. Fast positively-photoswitchable fluorescent protein for ultra-low laser power RESOLFT nanoscopy. Nature Methods, 12, 515-518, 2015. 3) Arai Y, et al. Spectral fingerprinting of indivisual cells visualized by cavity-reflection-enhanced light-absorption microscopy. PLoS ONE, 10, e0125733, 2015 4) Takai A, et al. Expanded palette of Nano-lanterns for real-time multicolor luminescence imaging. Proc. Natl. Acad. Sci. USA, 112, 4352-4356, 2015. 5) Saito K. & Nagai T. Recent progress in luminescent proteins development. Curr. Opin. Chem. Biol. 27, 46-51, 2015 6) Fukuda N, et al. Optical control of the Ca2+ concentration in a live specimen with a genetically encoded Ca2+ releasing molecu- lar tool. ACS Chemical Biology, 9, 1197-1203, 2014 7) Takemoto K, et al. SuperNova, a monomeric photosensitizing fluorescent protein for chromophore-assisted light inactivation. Scientific Reports, 3, 2629, 2013. 8) Saito K, et al. A luminescent protein for high-speed single-cell and whole-body imaging. Nature Communications, 3, 1262, 2012 9) Zhao Y, et al. An expanded palette of genetically encoded Ca2+ indicators. Science, 333, 1888-1891, 2011. 10)Horikawa K, et al. Spontaneous network activity visualized by ultra-sensitive Ca2+ indicators, yellow cameloen-Nano. Nature Methods 7, 729-732, 2010 11)Tomosugi W, et al. An ultramarine fluorescent protein with increased photostability and pH insensitivity. Nature Methods 6: 351-353, 2009 12)Matsuda T, et al. Direct measurement of protein dynamics inside cells using a rationally designed photoconvertible protein. Nature Methods 5:339-345, 2008 21 Laboratory of Clinical Pharmaceuticals Associate Professor (Clinical Professor) Yoshihiro MIWA +81-6-6879-6000 Clinical Associate Professor Masato KATSUURA +81-6-6879-6002 FAX +81-6-6879-5999 "The clinical pharmacology" is specialties of the pharmacy corresponding to the medical "clinical pharmacology" and can define pharmacokinetics / drug interactions / pharmacodynamics to study from the pharmacy field. The hospital pharmacy attached to the Osaka University School of Medicine shares important duties to support medical treatment through pharmaceutical products. The pharmacy that have a relation until the evaluation of manage- ment and the supply of medical supplies, the application to a human body and the effect, a side effect in each stage of the clinic use of medical supplies, and contacts with the medical settings directly of "the clinical pharmacology" practice it, and have many study materi- als. "The hospital practical pharmacy field" belonging to the pharmacy in that performs a study to aim at develop from the pharmacotherapy through a clinical case study and a study of the drug informatics study. Also, pharmacy school Pharm.D course (Osaka University) In cooperation with the field of / presenting part medicine studies and Pharm.D education promotion unit presenting part clinical pharma- cology research project, we wrestle with the study that aimed at the individualization appropriateness administration that assumed The blood level measurement in the pharmacy study pharmacokinetic study and genome pharmacology a base. We cross the medical supplies studying in our field till now to many divergences including the thing beyond the frame of the conventional medical supplies including high antibody medical supplies of the specificity derived from not only a small molecule compound and the physiologically active substance used widely but also cells. Some antimicrobial agents and anticancer drugs conduct the blood level measurement and conduct the analysis (TDM). It is necessary to evaluate poor effect on radiation exposure or environment to a healthcare worker to treat to medicine with carcino- genicity and teratogenicity like an anticancer drug. There is many that a previous finding is established anew by progress of a study instru- ment and the measuring equipment, and it is necessary to grasp the always latest study technique to evaluate it appropriately. The clinical pharmacology lecture is intended we estimate the problems by various clinics as a pharmacist in the viewpoint of the scientist, and to be settled. Therefore in our varying interest is interested, and demand the talented person who can wrestle positively. The anticancer drug preparation at the pharmacy oncology center Research topics 1) An adequacy study of the pharmacotherapy through the case study 2) An effect / adverse event manifestation research of the pharmacotherapy that assumed medical supplies prescription trend a base 3) Pharmacokinetic analysis by the development of the new blood level assay of the anticancer drug and inspection of the clinical signifi- cance 4) Effect on healthcare worker and environment investigation due to the radiation exposure of the anticancer drug 5) A pharmacokinetics-related genetic clinical genome pharmacology study 6) The inspection of the side effect of drug interactions manifestation risk 7) Development of the new hospital preparation of high quality 8) The elucidation of the acquisition of resistance mechanism of the molecular target medicine 9) The elucidation of the factor about molecular target adverse drug reactions manifestation Recent publications 1) Yamamoto T. et al., The effect of aprepitant and dexamethasone combination on paclitaxel-induced hypersensitivity reaction. EUROPEAN JOURNAL OF GYNAECOLOGICAL ONCOLOGY, 2016 in press 2) Naito M. et al., Hemoglobin values is the important factor for developing Hand-foot-syndrome in the Capecitabine regimen. Chemotherapy, 62 (1), 23-29, 2016 ࡔఈȄέͿϋΗΣσ࢛࣌ු྄ݟਓस͈ঀဥఠऔ. Palliative Care Research , 10(1), 107-112, 2015 (3 4) Machida H. et al., Correlation between Outbreaks of Multidrug-Resistant Pseudomonas Aeruginosa Infection and Use of Bronchoscopes Suggested by Epidemiological Analysis. Biological and Pharmaceutical Bulletin, 37(1), 26-30, 2014 5) Yabuno K. et al., High-dose, short-interval daptomycin regimen was safe and well tolerated in three patients with chronic renal failure. Clinical Pharmacology : Advances and Applications, 5, 161-166, 2013 ෝ̳͂ͥίυΈρخழ̵͙ࣣͩੜ༷ͬഩঊσΞΏΑΞθષ́̈́ثहࡩ̳ͥಕৣࢯ̦ͭस͈հڒུఈĭġ ໝତܰ (6 2013 ,498-491 ,(8)39 ,ڠอįġ֓ၷٳθ͈ 22 Laboratory of Experimental Genome Research Professor Masahito IKAWA +81-6-6879-8375 [email protected] Assistant Professor Yuhkoh SATOUH +81-6-6879-8375 [email protected] Assistant Professor Yoshitaka FUJIHARA +81-6-6879-8375 [email protected] Assistant Professor Haruhiko MIYATA +81-6-6879-8375 [email protected] Assistant Professor Masashi MORI +81-6-6879-8375 [email protected] Assistant Professor Daiji KIYOZUMI +81-6-6879-8375 [email protected] FAX +81-6-6879-8376 Professor Masahito IKAWA In the "post-genome project era", genetically modified Using gene knockout technology, we showed that testis- animals play a key role in basic investigations of molecular specific chaperones (CLGN, CALR3, and PDILT) are biology and providing human disease models. Our laboratory required for the quality control of the sperm membrane elucidates the mechanisms of mammalian reproductive protein ADAM3 in the endoplasmic reticulum (Nature. 1997; systems through genetically modified animals. JCI. 2010). The mutant mice were male infertile because We were the first in the world to produce the genetically their spermatozoa lacking ADAM3 could not migrate through modified mice that express green fluorescent protein (GFP) all the utero-tubal junction. However, the mutant spermatozoa over the body (Febs Lett. 1997) (Figure 1). were able to fertilize eggs when they were directly deposited into the oviducts (PNAS. 2012, 2013). Figure 1. GFP-expressing mice are highly useful for many types of Yet another tool enhanced by our laboratory is lentiviral research projects. Genetically (LV) vector-mediated genetic manipulation in vivo. We modified animals play a key role developed the placenta specific gene manipulation method by in basic investigations of molecu- transducing blastocyst stage embryos with LV vectors (Nat lar biology and providing Biotechnol. 2007). Using this technique, we are trying to human disease models. elucidate the mechanism of implantation and placentation (PNAS. 2011; BOR. 2016). :HHVWDEOLVKHGUDWHPEU\RQLFVWHP (6 FHOOVDQGJHQHUDWHG PRXVHļUDW FKLPHULF DQLPDOV :H ZRXOG OLNH WR XVH WKLV The green fluorescent mice are highly useful for many types animal model to study body/organ size control in vivo and it of research projects and in utilizing one of these animals, we may provide a method for the derivation of various organs labeled sperm with fluorescent proteins and visualized the IURP(6RUL36FHOOV Genes Cell. 2011; Sci Rep. 2016). fertilization process (Exp Anim. 2010; JCS. 2010, 2012) Our recent interest is the application of CRISPR/Cas9 (Figure 2). system for the generation of genetically modified animals to study fertilization, implantation, and placentation. We were successful in mice and rats using sgRNA/Cas9 expressing plasmids (Sci Rep. 2013; DGD. 2014; PNAS. 2016). Recently, we found that sperm calcineurin (PPP3CC/PPP3R2) is essen- tial for sperm motility and proposed the sperm calcineurin as a target for male contraception (Science. 2015). For more information, please visit our homepage (http://www.egr.biken.osaka-u.ac.jp/index.php). Figure 2. Mouse eggs (left) and sperm (right) labeled with fluorescent proteins. These gametes are used to visualize the fertilization process. Research topics 1) Analysis of molecular mechanisms involved in mammalian gametogenesis, fertilization, implantation and placentation 2) Development of new technologies to produce genetically modified animals (CRISPR/Cas9 system, lentiviral vectors, etc.) 3) Analysis of interspecies cell-cell communication in xenogenic chimeric animals Recent publications 1) Kato K. et al. Structural and functional insights into IZUMO1 recognition by JUNO in mammalian fertilization. Nat Commun. 7:12198(2016) 2) Miyata H. et al. Genome engineering uncovers 54 evolutionarily-conserved and testis-enriched genes that are not required for male fertility in mice. PNAS. 113: 7704-10(2016) ,VRWDQL$HWDO*HQHUDWLRQRI+SUWGLVUXSWHGUDWWKURXJKPRXVHĸUDW(6FKLPHUDVSci Rep. 6:24215 (2016) 4) Miyata H. et al. Sperm calcineurin inhibition prevents mouse fertility with implications for male contraceptive. Science. 350: 442-5 (2015) 23 Laboratory of Nano-Design for Innovative Drug Development Professor Koichi YAMANISHI +81-6-6877-4804 [email protected] Associate Professor Yasuo YOSHIOKA +81-6-6877-4919 [email protected] Professor Koichi YAMANISHI As the recent Ebola virus outbreak in Africa and worldwide influenza pandemic have powerfully demonstrated, society demands that vaccines for infectious diseases are developed, produced and distributed. To meet this strong demand, The Research Founda- tion for Microbial Diseases of Osaka University (BIKEN) and Research Institute for Microbial Disease, Osaka University (RIMD) have established a new research organization, The BIKEN Innovative Vaccine Research Alliance Laboratories, to promote the cooperative development of vaccines. In these cooperative laboratories, we developing new technologies to accumulate the basic information required for designing next-generation vaccines. Most protein antigens, such as non-living macromolecules or protein-subunit antigens, evoke only weak or undetectable adaptive immune responses. Therefore, to develop effective vaccines, it is necessary to develop vaccine adjuvants and antigen delivery carriers. In addition, to develop optimal vaccines for clinical applications, it is important to understand the mechanism of action of vaccines on the immune system in terms of efficacy as well as safety. In this regard, our research is focused on optimizing vaccines related to drug delivery systems and safety science. /DERUDWRU\RI1DQRGHVLJQ IRULQQRYDWLYHGUXJGHYHORSPHQW 2XUUHVHDUFKLVIRFXVHGRQRSWLPL]LQJYDFFLQHV UHODWHGWRGUXJGHOLYHU\V\VWHPVDQGVDIHW\VFLHQFH 'HYHORSPHQWRIDQWLJHQ 'HYHORSPHQWRIYDFFLQH GHOLYHU\FDUULHUVDQGDGMXYDQWV DGMXYDQWXVLQJFRPSUHKHQVLYH XVLQJQDQRWHFKQRORJ\ VFUHHQLQJPHWKRGV 'HYHORSPHQWRIYDFFLQHVIRULQIHFWLRXVGLVHDVHV XVLQJRXUGHYHORSHGDGMXYDQWVDQGGHOLYHU\FDUULHUV Research topics 1) Development of vaccine adjuvant using comprehensive screening methods 2) Development of antigen delivery carriers and adjuvants using nanotechnology 3) Development of vaccines for infectious diseases using our developed adjuvants and delivery carriers Recent publications 1) Hirai T, Yoshioka Y, Metal nanoparticles in the presence of lipopolysaccharides trigger the onset of metal allergy in mice. Nat Nanotechnol. 2016 May 30. 2) Yoshida T, Yoshioka Y, et al. Intranasal exposure to amorphous nanosilica particles could activate intrinsic coagulation cascade and platelets in mice. Part Fibre Toxicol. 10:41, 2013. 3) Yamashita K, Yoshioka Y et al. Silica and titanium dioxide nanoparticles cause pregnancy complications in mice. Nat Nano- technol. 6:321-8, 2011. 4) Kayamuro H, Yoshioka Y, et al. Interleukin-1 family cytokines as mucosal vaccine adjuvants for induction of protective immunity against influenza virus. J Virol. 84:12703-12, 2010. 24 Laboratory of Applied Pharmacognosy Associate Professor Kyoko TAKAHASHI +81-6-6879-8160 [email protected] Specially Appointed Assistant Professor Kayoko TAKAURA +81-6-6879-8160 [email protected] FAX +81-6-6879-8160 Challenge of drug treatment in 21st Century is “proper use of medicine” and “tailored medication”. Traditional medicine has been practiced over thousands of years, and is currently utilized worldwide. Combination therapy of Western and Eastern medicine is effective for the diseases whose cause is not single, but complicated. Traditional medicines have been developed and originated from crude drugs. All the medicinal studies are based on the identification and quality assessments of medicinal materials. Traditional medicines such as Kampo and Traditional Chinese Medi- Scientific Tools for Studying cines are the combination drugs, and considered that several compounds ZĞƐĞĂƌĐŚWƌŽũĞĐƚƐŽŶ Secure Quality Associated with work interactively, therefore it’s difficult to figure out their mechanisms. ŶǀŝƌŽŶŵĞŶƚĂů Efficacy and Toxicity ^ƵƐƚĂŝŶĂďŝůŝƚLJ However, accumulated evidence for traditional medicines may expand medical practice and lead new treatment method for intractable diseases. Most of the crude drugs in Japan are imported, and thier quality associ- ated with efficacy and toxicity depends on various factors. There is also +HUE7HD In vitro Study of Multi-element an urgent need for development of new or alternative drug resources. To ĂŶĚĞůŝŽŶ'ƌŽǁŶŝŶKƐĂŬĂ Cultured Cardiac Determination, secure quality of traditional medicine, it’s important to establish the hŶŝǀĞƌƐŝƚLJĂƐ ǀĂůƵĂƚŝŽŶdŽŽů Cells Metabolomics specific evaluation model for traditional medicine and to verify the Maintenance of Resources and effectiveness. To clarify scientifically the mechanism and the efficacy of Qualitative Evaluation those medicines are required for the evidence based medicine (EBM). Programmed of Crude Drugs Furthermore, historical literature and crude drug specimens are Learning for Kampo- Pharmacy valuable source to expand clinical usage of traditional medicine. Histor- EBM and High Clinical cal medical records are the accumulated knowledge based on the past Application of Quality Medical Traditional experiences, and really valuable data which are equivalent to enormous Care Medicines clinical trial data including ethnic differences. The historical crude drug specimens possibly become a trigger to reveal the past medical practice 7RFKLPRWR 7HQNDLGR &R/WG since those samples are the voucher specimens to show the origins of Scientific Studies on Cultural Properties / those drugs. By analyzing them, it might be possible to obtain insights Natural History䞉 for exploring the standardized quality and efficacy of crude drugs and Ethno-medicines development of cultivated type with high quality. Osaka University Scientific Studies on The Database of Ethno-medicines / retains “Koan Ogata’s medicine chest”, and any other valuable crude Cultural Properties Medicinal Resources drug samples, one of key areas in our researches is to investigate these historically valued sources. In our research laboratory, we investigate about sustainable supply of crude drugs with high quality, and we also aim to utilize the accumulated knowledge based on the past medical practice by evaluating historical data. Our researches can contribute to EBM and high-quality medical practice by accumulating the scientific evidence related to traditional Koan Ogata’s medicine chest medicine. Owned by Osaka University Research topics 1) Sustainable use and quality control of crude drugs 2) Establishment of the database of ethno-medicines / medicinal resources 6FLHQWLILFDVVHVVPHQWRIVDIHW\DQGHIILFDF\RIWUDGLWLRQDOPHGLFLQHV (VWDEOLVKPHQWRIFRQVRUWLXPDQGWKHVL[WKLQGXVWULDOL]DWLRQRIKHUEDOPHGLFLQHSURGXFWLRQ 6FLHQWLILFDQDO\VLVRI³.RDQ2JDWD¶VPHGLFLQHFKHVW´ Recent publications 6KHWHZ\$HWDO0LWRFKRQGULDOGHIHFWVDVVRFLDWHGZLWKȕDODQLQHWR[LFLW\ UHOHYDQFHWRK\SHUEHWDDODQLQHPLDMolecular and Cellular Biochemistry (2016) 416 2) Oguri K. et al., Longgu (Fossilia Ossis Mastodi) alters the profiles of organic and inorganic components in Keishikaryukot- suboreito. Journal of Natural Medicines (2016) 70 6FKDIIHU6:HWDO,PSDLUHGHQHUJ\PHWDEROLVPRIWKHWDXULQHGHILFLHQWKHDUWAmino Acids (2016) 48 6KLPDGD.HWDO5ROHRI526SURGXFWLRQDQGWXUQRYHULQWKHDQWLR[LGDQWDFWLYLW\RIWDXULQHAdv Exp Med Biol 803 6FKDIIHU6:HWDO(IIHFWRIWDXULQHDQGSRWHQWLDOLQWHUDFWLRQVZLWKFDIIHLQHRQFDUGLRYDVFXODUIXQFWLRQAmino Acids 46 6KLPDGD.HWDO(VWDEOLVKPHQWRIDPHWKRGRORJ\IRULGHQWLI\LQJ3DHRQLDH5DGL[EDVHGRQPHWDOORPLFDQDO\VLVJ Nat Med 68 6KLPDGD.HWDO7DXULQHDVDPDUNHUIRUWKHLGHQWLILFDWLRQRIQDWXUDOCalculus Bovis and its substitutes. Adv Exp Med Biol 776 25 Department of Innovative Clinical Trials and Data Science Professor Haruko YAMAMOTO +81-6-6833-5012 [email protected] Professor Toshimitsu HAMASAKI +81-6-6833-5012 toshi.hamasaki@ ncvc.go.jp During the last several years, clinical research infrastructure for advancing medical product development including both pharmaceu- tical products/medical devices has been strengthen, Such intensive efforts, however, still fall short of expected levels of reduction in resource including overall time and cost for the medical product development. Even innovative technologies of the 21st century has not contributed so much for improved success in medical product development, and successful regulatory approval rate remain stagnant: 18% of the programs originating from nonclinical studies and for 22% of the projects initiated from Phase I studies have never been high. In this modern area where new medical products are simultaneously, globally developed in multi-regions, such stagnation constitutes problems not only for our country where drug/device lag issues become more apparent but are shared world- wide. Currently, the importance lies in streamlining the foundations and infrastructure of a series of processes for medical products from basic research studies though clinical development to the commercialization and utilization of the products/devices at the front of medical practices, as well as in promoting human resource cultivations in an integral fashion. On top of such promotion, challenges strongly desired by society exist for promptly providing people in our society with cutting-the-edge medical care, solutions for unmet medical needs for personalized medicines evolving on the global level and rare/intractable diseases, and for promoting “medical innovation” to facilitate the advancement of medical service and revolution of the medical system at the same time. In this field, our research contribute to earlier achievement of a healthy society through the following efforts: 1. cultivation of human resources with high professionality and capabilities for delivering the professionality into practice with high ethical perspectives and a research mindset; 2. research on methodologies aiming at highly-productive, cost-effective medical product development and accel- eration of processes from basic research to commercialization of medical products through delivering information; and 3. faster achievement of medical innovations through worldwide research/education network. Notably, we host an environment within the same facility by employing both basic researchers devoted for pharmaceutical product/medical device developments and highly professional clinicians capable of delivering advanced medical care, working closely in tandem for commitment towards the entire process of clinical research programing to implementation of such programs. Altogether, we can provide highly advantageous environment for cultivating the professionality for clinical development of pharmaceutical products/medical devices. A further possibility lies ahead of the research life in this field for broader capabilities such as clinical development specialists on pharmaceutical product/medical device, project managers for clinical studies, and data manager specialists in regulatory affairs, and medical statisticians. Research topics 1) Regulatory science-based approach for evaluating safety and efficacy of new medical products 2) Risk-based approach for monitoring and improving quality of clinical studies 3) Statistical methodologies for streaming translational research 4) Statistical methodologies for complex clinical trial design and analysis 5) Data-oriented approach for profiling patients characteristics via big data Recent publications 1) Hamasaki T, Asakura K, Evans S, Sugimoto T, Sozu T. Group sequential strategies for clinical trials with multiple co-primary endpoints. Stat Biopharm Res 7, 36-54, 2015 (Most Accessed Articles and Most Cited Articles). 2) Koga M，Toyoda K，Kimura K，Yamamoto H，Sasaki M，Hamasaki T，Kitazono T，Aoki J，Seki K，Homma K， Sato S，Minematsu，K，on behalf of the THAWS investigators. Thrombolysis of acute wake-up and unclear-onset strokes with alteplase at 0.6 mg/kg (THAWS) trial. Int J Stroke 2014; 9: 1117-1124. 3) Asakura K, Hamasaki T, Sugimoto T, Hayashi K, Evans SR, Sozu T. Sample size determination in group-sequential clinical trials with two co-primary endpoints. Statist Med 2014; 33: 2897-2913. 4) Sugimoto T, Sozu T, Hamasaki T, Evans SR. A logrank test-based method for sizing clinical trials with two co-primary time- to-events endpoints. Biostatistics 2013; 14, 409-421, 2013. 5) Hamasaki T, Sugimoto T, Evans SR, Sozu T. Sample size determination for clinical trials with co-primary outcomes. Expo- nential event-times. Pharm Stat 2013; 12: 28-34. (Most Accessed Papers Published in 2012 & 2013) 26 Laboratory of Biomedical Engineering Professor Tetsuji YAMAOKA +81-6-6833-5012 (ex2637) [email protected] Professor Tetsuji YAMAOKA We have been proposing new advanced medical devises and systems as follows in order to understand and control cerebral and cardiovascular diseases and to contribute to human health and well-being. Our Lab mainly consists of engineers and collaborates with clinicians, basics medical scientists, and of other scientific fields such as biochemistry and cell biology inside and outside our Center [1] DNCS (Drug-Navigated Clearance System) Some cytokines and inflammatory mediators are considered to be pathogeneses of rheumatoid arthritis (RA), which are referred to as rheumatoid factors (RF). We have developed a novel therapeutic strategy, drug-navigated clearance system (DNCS), which leads the autoantibody, RF, toward the hepatic LDL receptors (LDLRs) by use of a newly designed drugs, "navigator". Protein A that bind to the target antibody (RF) was chemically conjugated to apolipoprotein E (ApoE) which directly bind to the hepatic LDLR and dextran sulfate (DexS) which binds to LDL and then indirectly accumulate to the liver. The conjugation reaction was monitored on SDS-PAGE. In vitro uptake of fluorescently labeled IgG by HepG2 was greatly increased in the presence of naviga- tor molecule. In addition, the elevated RF level of the RA model rat was reduced immediately after the navigator injection and last for about 2 days. [2] Acellular Small Diameter Long Bypass Grafts Researchers have attempted to develop efficient antithrombogenic surfaces, and yet small-caliber artificial vascular grafts are still unavailable. Here, we demonstrate the excellent patency of tissueengineered small-caliber long-bypass grafts measuring 20e30 cm in length and having a 2-mm inner diameter. The inner surface of an acellular ostrich carotid artery was modified with a novel heterobifunctional peptide composed of a collagen-binding region and the integrin a4b1 ligand, REDV. Six grafts were transplanted in the femoralefemoral artery crossover bypass method. Animals were observed for 20 days and received no antico- agulant medication. No thrombogenesis was observed on the luminal surface and five cases were patent. In contrast, all unmodi- fied grafts became occluded, and severe thrombosis was observed. The vascular grafts reported here are the first successful demon- strations of short-term patency at clinically applicable sizes. Research topics 1) DNCS (Drug-Navigated Clearance System) 2) Acellular Small Diameter Long Bypass Grafts 3) MRI tracking of living stem cells 4) Injectable Hydrogels for Myocardial Infarction Treatment 5) Pluripotent cell separation using "cell rolling column" Recent publications 1) Y. Kambe et al., Development of a FRET-based recombinant tension sensor to visualize cell–material interactions, J Mater. Chem B, 4, 649-655 (2016). 2) A. Mahara et al., Tissue-Engineered Acellular Small Diameter Long-bypass Grafts with neointima-inducing activity, Biomaterials, 58, 54-62 (2015) 3) S. Kakinoki et al., Accelerated tissue integration into porous materials by immobilizing basic fibroblast growth factor using a biologically safe three-step reaction, J. Biomed. Mater. Res. Part A, 103(12):3790-3797 (2015) 4) P. H. Liem et al., Preparation of inactivated human skin using high hydrostatic pressurization for full-thickness skin recon- struction, PLoS ONE, PLoS ONE 10(7): e0133979 (2015). 27 Laboratory of Vaccine Materials Professor Jun KUNISAWA +81-72-641-9871 [email protected] FAX +81-72-641-9872 Professor Jun KUNISAWA The aims of Kunisawa lab are to understand the host immunity controlled by gut environment such as commensal bacteria and diets and to apply the basic findings to the development of vaccines, medicines, and functional foods. For the vaccine development, Kunisawa lab has already identified unique molecules derived from commensal and pathogenic bacteria for the antigen delivery and adjuvant for mucosal (oral and nasal) vaccine. They have also tried to identify and design new and effective vaccine antigen. Gut Environment for the Control Host Immunity and As diets, Kunisawa lab focuses on oils and vitamins. As the basic research, they have examined for the Development of Vaccine and Immunotherapy immunologic functions of dietary oils as a source of teiD dna noitirtun slasnemmoC Pathogens lipid mediators and have identified some good Macrophage mediators for the control of allergic and inflamma- Basophil tory diseases. They are currently trying to translate them to the development of functional foods and medicine. In vitamin project, Kunisawa lab has Mast cell reported specific functions of some vitamins Vitamins, lipids, etc… (vitamin A and B family) as a key regulator of T DC particular immune responses. Finally, they have IgA PC performed epidemiologic studies using healthy Stromal cell Eosinophil individuals with different habit (food and exercise) and patients with allergic and inflammatory Interaction among “Diet-Commensal-Host Immunity” diseases. Infection/immune disease By combing both basic and epidemiologic Ὁ Immunosurveillance researches, Kunisawa lab aims to provide cutting- Ὁ Immune diseases (inflammation and allergy) edge information for human health and diseases and to develop prospective vaccines, medicines, and Development of new type of vaccine, medicine, functional functional foods. Research topics 1) Vaccine and drug development for infectious diseases 2) Diet-mediated immune regulation for the development of functional foods 3) Commensal bacteria in the health and diseases 4) Epidemiology for human health and diseases Recent publications 1) J. Kunisawa et al, Mode of bioenergetic metabolism during B cell differentiation in the intestine determines the distinct requirement for vitamin B1. Cell Rep 13: 122-31, 2015 -.XQLVDZDHWDO'LHWDU\ȦIDWW\DFLGH[HUWVDQWLDOOHUJLFHIIHFWWKURXJKWKHFRQYHUVLRQWRHSR[\HLFRVDWHWUDHQRLFDFLG in the gut. Sci Rep 5: 9450, 2015 3) Y. Kurashima et al, The enzyme Cyp26b1 mediates inhibition of mast cell activation by fibroblasts to maintain skin-barrier homeostasis, Immunity 40: 530-41, 2014 4) J. Kunisawa et al, Microbe-dependent CD11b+ IgA+ plasma cells in early-phase robust intestinal IgA responses in mice, Nat Commun 28 Laboratory of Innovative Antibody Engineering and Design Professor Satoshi NAGATA +81-72-641-9883 [email protected] Associate Professor Tomoko ISE +81-72-641-9883 [email protected] +81-72-641-9884 Professor Satoshi NAGATA Our group studies antibody-based therapeutics. One of our focuses is developing a methodology for production of structure-based antibody libraries. This new type of library would enable us to utilize multiple, distinct structures displayed on a single target antigen. For example, from the libraries, we will be able to isolate a set of two different monoclonal antibodies that can bind to a single antigen simultaneously. Another focus of our study is therapeutic manipulation of the interaction between cancer cells and host immune system with antibodies. We are studying many members of receptor families whose epitopes are structurally similar to each other. For examples, we study the tumor necrosis factor receptor superfamily (TNFRSF) and the immunoglobulin receptor superfamily. Research topics 1) Development of therapeutic antibodies against membrane proteins including virus envelope proteins. 2) Analysis and therapeutic manipulation of the interaction between cancer cells and host immune system. Recent publications 1) Li H, Borrego F, Nagata S, Tolnay M (2016) Fc receptor-like 5 expression distinguishes two distinct subsets of human circu- lating tissue-like memory B cells. The Journal of immunology, 196, 4064-4074. 2) Bin Dhuban K, d'Hennezel E, Nashi E, Bar-Or A, Rieder S, Shevach EM, Nagata S, Piccirillo CA (2015) Coexpression of TIGIT and FCRL3 Identifies Helios+ Human Memory Regulatory T Cells. The Journal of immunology 194, 3687-3696. 3) Terrier B, Nagata S, Ise T, Rosenzwajg M, Pastan I, Klatzmann D, Saadoun D, Cacoub P (2014) Clonal CD21-/lo marginal zone-like B cells in HCV-related lymphoproliferation highly express Fc receptor-like 5 protein on the cell surface and are specifically killed by anti-FCRL5 immunotoxins, Arthritis & Rheumatism 66, 433-43. 4) Wysocka M, Kossenkov AV, Benoit BM, Troxel AB, Singer E, Schaffer A, Kim B, Dentchev T, Nagata S, Ise T, Showe LC, Rook AH (2014) CD164 and FCRL3 Are Highly Expressed on CD4+CD26- T Cells in Sézary Syndrome Patients, Journal of Investigative Dermatology 134, 229-236. 29 Laboratory of XNA Screening and Design Professor Yoshihiro YONEDA +81-72-641-9811 [email protected] Researche Yuuya KASAHARA +81-72-641-9882 [email protected] It becomes possible to use Xeno nucleic acid (XNA) as a functional and key element of oligonucleotide therapeutics or nucleic acid medicines. For example, ‘Kynamro’ is an antisense drug recently approved by FDA in USA. The antisense drugs hybridize with an mRNA of target gene in a sequence specific manner and result in degradation of the target mRNA to inhibit the disease- related protein production. ‘Macugen’, another type of nucleic acid medicines, is an aptamer drug and it was approved not only in USA but also in Japan. The aptamer drugs bind to a disease-related protein and inhibits its function. Thus, there are various types of nucleic acid medicines that can target DNA, RNA, proteins or any other molecules in vivo. They are expected to be a “magic bullet” for intractable and/or rare diseases that cannot be solved by conventional therapy. In our laboratory, we have designed and synthesized several new XNAs and developed a screening technique to select functional nucleic acid medicines. To improve the effect of antisense drugs, biological stability in vivo, cell penetration ability and safety as a medicine should be carefully considered. In addition, binding affinity to a target protein, inhibitory activity and retentivity in vivo could be important factors for nucleic acid aptamers. We have synthesized new XNAs of which nucleobases, sugars, and/or phosphodiester linkages are chemically modified. We also developed new functional XNAs that have an ability to change their properties spatiotemporally in response to an external-stimulus. These XNAs could be useful to improve drug efficacy and reduce doses and side effects when they are introduced into nucleic acid medicines. Recently, we also successfully achieved the synthesis of XNAs that can work as a fluores- cent probe. This made it possible to detect Various kinds ofartificialoligonucleotide mismatch base pair in nucleic acid sequences by the difference of fluorescence intensity. Antisense In sequence screening study, we aim to NucleicͲacid RNAi oligonucleotide select highly active sequences of antisense aptamer drugs or nucleic acid aptamers efficiently siRNA Ex.䠅Kynamro Ex.䠅Macugen from sequence libraries. For the nucleic acid miRNA aptamers-based drug discovery, it is essential to introduce XNAs into aptamers; however, Transcription Translation conventional polymerases have no ability to incorporate XNAs into oligonucleotides. Thus, we prepared considerable numbers of RNA splicing modified polymerases that have an ability to DNA PreͲmRNA mRNA Protein handle XNAs. In this way, we have consistently carried out a study of XNAs, from the design, synthesis Decoy Spliceswitching and screening to the biological application, for nucleicacid oligonucleotides the purpose of XNA-based drug discovery. Research topics 1) Design and synthesis of XNAs to improve the ability of nucleic acid medicines. 2) Design and synthesis of XNAs that change their function in response to an external stimulus. 3) Preparation of modified polymerases to incorporate XNAs into oligonucleotides effectively. 4) Development of a highly efficient screening method for nucleic acid medicines.. 5) Synthesis and application of fluorescent nucleic acids. 6) Development of nucleic acid medicines to treat refractory rare diseases. Recent publications 1) Okuda T., Mori S., Kasahara Y., Morihiro K., Ikejiri M., Miyashita K., Obika S., Tetrahedron Lett., 2016, 57, 3129-3132. 2) Hoshino H., Kasahara Y., Fujita H., Kuwahara M., Morihiro K., Tsunoda S., Obika S., Bioorg. Med. Chem. Lett., 2016, 26, 530-533. 3) Morihiro K., Hoshino H., Hasegawa O., Kasahara Y., Nakajima K., Kuwahara M., Tsunoda S., Obika S., Bioorg. Med. Chem. Lett., 2015, 25, 2888-2891. 4) Morihiro K., Hasegawa O., Mori S., Tsunoda S., Obika S., Org. Biomol. Chem., 2015, 13, 5209-5214. 5) Mori S., Morihiro K., Kasahara Y., Tsunoda S., Obika S., Chemosensors, 2015, 3, 36-54. 30 Laboratory of Drug Target Discovery Professor Haruhiko KAMADA +81-72-641-9814 [email protected] FAX +81-72-641-9817 Professor Haruhiko KAMADA A lot of refractory disorders, including cancer, and inflammatory diseases, are characterized by alterations in the molecular distri- bution of vascular structures, presenting the opportunity to use biopharmaceutics, such as monoclonal antibodies for clinical thera- pies. This strategy, often referred to as "vascular targeting", has been promising in promoting the discovery and development of selective biological drugs to regulate angiogenesis-related diseases such as refractory malignances. Various experimental approaches have been utilized to discover secreted molecules and accessible vascular markers of health and disease at the protein level. Our group has developed a new proteomics technology to identify and quantify accessible vascular proteins in normal organs and at disease sites. Our developed methodology relies on the perfusion of animal models with suitable ester derivatives of biotin, which react with the primary amine groups of proteins as soon as the molecules are attached. This presentation reports biomedical applications based on vascular targeting strategies, as well as dĂƌŐĞƚŽŶƉůĂƐŵĂŵĞŵďƌĂŶĞ methodologies that have been /DEHOLQJFHOOVXUIDFHSURWHLQV used to discover new vascular $QWLERG\'UXJ ZLWKXQLTXHFKHPLFDODJHQW targets. The identification of antigens located in the stromal tissue of pathological blood vessels may provide attractive targets for the development of y +DUGO\ODEHOLQJ antibody drugs. This method will WKHLQQHUSURWHLQV also provide an efficient discov- 3URWHLQVH[SUHVVLQJ 3URWHLQVH[SUHVVLRQJ ery target that could lead to the 2QSODVPDPHPEUDQH F\WRSODVP development of novel antibody WƵƌŝĨŝĐĂƚŝŽŶĂŶĚŝĚĞŶƚŝĨŝĐĂƚŝŽŶŽĨĐĞůů drugs. %LQGLQJIRUQHXWUDOL]DWLRQ 8QERXQGWRUWDUJHW ŵĞŵďƌĂŶĞƚĂƌŐĞƚƐƵƐŝŶŐƉƌŽƚĞŽŵŝĐ ĂŶĂůLJƐŝƐ Research topics 1) Development of essential technology for drug target discovery 2) Drug delivery system for active targeting against the drug target molecule Recent publications 1) Arai M, Kawachi T, Kotoku N, Nakata C, Kamada H, Tsunoda S, Tsutsumi Y, Endo H, Inoue M, Sato H, Kobayashi M., Furospinosulin-1, Marine Spongean Furanosesterterpene, Suppresses the Growth of Hypoxia-Adapted Cancer Cells by Bind- ing to Transcriptional Regulators p54(nrb) and LEDGF/p75., Chembiochem., 17(2):181-189, 2016 2) Li J, Inoue J, Choi JM, Nakamura S, Yan Z, Fushinobu S, Kamada H., Kato H, Hashidume T, Shimizu M, Sato R.: Identifica- WLRQRIWKH)ODYRQRLG/XWHROLQDVD5HSUHVVRURIWKH7UDQVFULSWLRQ)DFWRU+HSDWRF\WH1XFOHDU)DFWRUĮ-%LRO&KHP 290(39): 24021-24035, 2015 3) Kamada H, Taki S, Nagano K, Inoue M, Ando D, Mukai Y, Higashisaka K, Yoshioka Y, Tsutsumi Y, Tsunoda S.: Generation and characterization of a bispecific diabody targeting both EPH receptor A10 and CD3. Biochem Biophys Res Commun. 456(4):908-912, 2015 4) Taki S, Kamada H, Inoue M, Nagano K, Mukai Y, Higashisaka K, Yoshioka Y, Tsutsumi Y, Tsunoda S.: A Novel Bispecific Antibody against Human CD3 and Ephrin Receptor A10 for Breast Cancer Therapy., PLoS One., 10(12):e0144712, 2015 5) Schliemann C, Roesli C, Kamada H, Borgia B, Fugmann T, Klapper W, Neri D.: In vivo biotinylation of the vasculature in B cell lymphoma identifies BST-2 as a target for antibody-based therapy. Blood, 115(3): 736-744, 2010 6) Kamada H., Fugmann T., Neri D., Roesli C. : Improved protein sequence coverage by on resin deglycosylation and cysteine modification for biomarker discovery. , Proteomics, 9(3):783-787, 2009 31 Laboratory of Computational Drug Discovery and Development Professor Kenji MIZUGUCHI +72-641-9890 [email protected] FAX +72-641-9881 http://mizuguchilab.org Professor Kenji MIZUGUCHI We are based in the National Institutes of Biomedical Innovation, Health and Nutrition, Japan and carry out bioinformatics and computational biology research into drug discovery and development. Our research ultimately aims to identify novel drug targets or drug candidates through the modeling and simulation of biological systems based on real-life experimental data. We develop software and databases to assist these processes and we also collaborate closely with a wide range of experimental groups. Several reasons can be attributed to costly late-phase attritions in drug development but the “wrong target” and the “wrong compound” are the main culprits. The former problem manifests itself in the poor correlation between in vitro and in vivo assay results and the difficulty in translating animal models to humans. Computational methods can improve target identification and validation at an early stage of drug development by providing a better understanding of the biology of the target network. We have developed TargetMine, an integrated data 0HWKRGVGHYHORSPHQW 3UHGLFWLRQDQGGDWDDQDO\VLV warehouse for assisting early stage drug 7DUJHWLGHQWLILFDWLRQ discovery and we aim to establish systems approaches to drug discovery. TargetMine 'DWDZDUHKRXVH 3XEOLFGDWDEDVHV 3URWHLQ and other computational biology techniques, &RPSRXQG utilizing information about protein sequence, $SSOLFDWLRQ structure and interactions, have been applied *HQRPH 'LVHDVH to specific drug discovery projects, including 1HZ hepatitis C virus (HCV) infection and breast 3UHGLFWLRQRIVWUXFWXUH 0RGHOLQJ IXQFWLRQDQGLQWHUDFWLRQ +\SRWKHVLV cancer, and led to several novel discoveries. )HHGEDFN To address the issue of the “wrong compound”, $QWLJHQ we develop computational resources for predicting ([SHULPHQWDO toxicity and pharmacokinetics of drug candi- $QWLERG\ YDOLGDWLRQ dates, as well as improving in silico screening In silico screening methods. We have also developed methods &DQGLGDWH 1RYHOWDUJHWV for predicting protein structure, function and FRPSRXQG 1RYHOGUXJV interaction. All these tools are being utilized in specific drug discovery projects such as the development of new vaccines and anti-cancer drugs. Research topics 1) Analysis of large-scale experimental data and the development of integrated databases for target discovery and safety prediction. 2) Prediction of protein-protein or protein-small molecule interactions, modeling of their impact on phenotypic responses and applications to drug design. Recent publications 1) Tsuchiya Y, et al. Ligand-induced Ordering of the C-terminal Tail Primes STING for Phosphorylation by TBK1. EBioMedicine, 9:87-96, 2016. 2) Chen YA, et al. Integrated pathway clusters with coherent biological themes for target prioritisation. PLoS One, 9:e99030 , 2014. 3) Yoshimaru T, et al. Targeting BIG3-PHB2 interaction to overcome tamoxifen resistance in breast cancer cells. Nat Commun, 4:2443, 2013. 4) Nystrom-Persson J, et al. Toxygates: interactive toxicity analysis on a hybrid microarray and linked data platform. Bioinformatics, 29:3080-3086, 2013. 32 Laboratory of Primate Research Professor Yasuhiro YASUTOMI +81-29-837-2053 (Tel & Fax) [email protected] Professor Yasuhiro YASUTOMI There are various important processes that must be carried out for the development of new drugs based on results of basic medical science research. Among the various processes, experiments using animals to determine the effectiveness and toxicity of new drugs are very important. Due to the dramatic progress in genetic engineering technology, comprehensive analyses have recently been carried out using genetically modified animals, particularly mice, with deficiencies or overexpression of genes. Although such experimental animals are very important for investigating functions and other aspects of genes, it is not clear whether results obtained by using experimental animals are applicable to humans for the development of new drugs. Primates have long been used as experimental animals that are similar to humans. Cynomolgus monkeys (Macaca fascicularis), unlike other animals, have various characteristics similar to those of humans such as a long life, single pregnancy and the same menstrual cycle. They are also valuable as experimental animals because they are affected by the same infectious diseases that occur in humans. Our research laboratory is in Tsukuba Primate Research Center, National Institutes of Biomedical Innovation, Health and Nutrition (NIBIOHN), the only primate center in Japan. The cynomolgus monkeys in our center are kept in an SPF facility, one of the few such facilities for cynomolgus monkeys in the world, and they are also genetically controlled as a closed colony. Research using a range of various experimental animals from mice to primates is being carried out in our laboratory. dilative myopathy macular degeneration iPS brain senile plaques AIDS Research topics 1) ＜Research on spontaneously occurring diseases＞ Cynomolgus monkeys in our center are being maintained as various models of spontaneously occurring diseases (including macular degeneration, dilative cardiomyopathy, Alzheimer’s disease, endometriosis and diabetes mellitus), and research on the onset and progression of those diseases as well as research for the development of therapeutic drugs are being carried out. 2) ＜Disease models＞ Research is being carried out using experimentally produced disease models to analyze the diseases and develop drugs and vaccines. In facilities for research on infectious diseases using primates, including ABSL2 and ABSL3 facilities, experiments are being carried out using more than 300 animals. Recent publications 1) Tsujimura Y., Yasutomi Y. Allergy vaccines using a Mycobacterium-secreted antigen, Ag85B, and IL-4 an antagonist. Vaccine Design 2016:pp723-738. 2) Tsujimura Y, Inada H, Yoneda M, Fujita T, Matsuo K. and Yasutomi Y. Effects of Mycobacteria major secretion protein, Ag85B, on allergic inflammation in the lung. PLos One 2014;9: E-pub 3) Watanabe K., Matsubara A, Kawano M, Mizuno S, Okamura T, Tsujimura Y, Inada H, Nosaka T, Matsuo K. and Yasutomi Y. Recombinant Ag85B vaccine by taking advantage of characteristics of human parainfluenza type 2 virus vector showed Mycobacteria-specific immune responses by intranasal immunization. Vaccine 2014:32;1727-1735. 4) Shimozawa N, Ono R, Shimada M, Shibata H, Takahashi I, Inada H, Takada T, Nosaka T, Yasutomi Cynomolgus monkey induced pluripotent stem cells established by using exognous genes derived from the same monkey species. Differentiation 2013:85;131-139. 33 Laboratory of Medicinal Plant Resources Professor Nobuo KAWAHARA +81-29-838-0571 [email protected] FAX +81-29-838-0575 Professor Nobuo KAWAHARA As best represented by Kampo medicine, medicinal plants have been used as ingredients for pharmaceutical products from ancient times. The Research Center for Medicinal Plant Resources, the only comprehensive research center in Japan for medicinal plants, performs research and development on technology related to the cultivation and breeding of medicinal plants, chemical and biological evaluations, and cultivates and preserves more than 4,000 species and groups of medicinal plants at three divisions in Japan suitable to the vegetation of the area, to supply seeds and seedlings and offer guidance on cultivation technology to various research institutes. Furthermore, we disclose ‘Comprehensive Medicinal Plant Database’ including the above information. Laboratory Medicinal Plant Resources studies on development of new technology for preservation of seeds and seedlings, breed- ing of new varieties, hydroponic cultivation of medicinal plants and providing related information. Recently, we are constructing a large-scale extract library based on plant resources in Japan and abroad, so that plants can be used as a resource for drug discov- ery. This project is aimed at utilizing the extract library to screen for various biological activities and find innovative seeds for pharmaceutical products. Research topics 1) Study on discovery of innovative seeds for pharmaceuticals by using ‘Medicinal Plant Extract Library’ 2) Study on information and correlation analysis of ‘Comprehensive Medicinal Plant Database’ 3) Study on breeding of high quality and high productivity of new medicinal plant variety by using ‘Medicinal Plant Factory’ 4) Study on establishment of ‘Medicinal Plant EST library’ and utilization of EST information Recent publications 1) Yahara Y, et al., Pterosin B prevents chondrocyte hypertrophy and osteoarthritis in mice by inhibiting Sik3. Nature Communications, 7, 10959 (2016). 2) Wahyuni T. S, et al., Inhibition of Hepatitis C Virus Replication by Chalepin and Pseudane IX Isolated from Ruta angustifolia Leaves. Fitoterapia, 99 (12), 276-283 (2014). 3) Fuchino H, et al., Relationship between the inhibitory effect of ginger (Zingiber officinale Roscoe) on nitric oxide production and the drying conditions after harvest, Jpn. J. Food Chem. Safety, 21 (1) 8-14 (2014). 4) Adianti M, et al., Anti-hepatitis C virus compounds obtained from Glycyrrhiza uralensis and other Glycyrrhiza species. Microbiol Immunol. 58 (3), 180-187 (2014). 5) Fuchino H, et al., Two New Labdane Diterpenes from Fresh Leaves of Leonurus japonicus and Their Degradation during Drying. Chem., Pharm., Bull., 61 (5), 497-503 (2013). 34 Laboratory of Stem Cell Regulation Professor Kenji KAWABATA +81-72-641-9815 [email protected] FAX +81-72-641-9816 Professor Kenji KAWABATA Embryonic stem (ES) cells and induced pluripotent stem (iPS) cells show the characteristics of self-renewal and pluripotency. In our laboratory, functional cells, such as hematopoietic cells, immune cells, and vascular endothelial cells, are efficiently differenti- ated from stem cells. Our objectives are to use these differentiated cells for the development of novel drug screening systems and in vitro disease models. 'UXJGHYHORSPHQWE\XVLQJVWHPFHOO GHULYHGIXQFWLRQDOFHOOV 0DVWFHOOVGLIIHUHQWLDWHG IURPL36FHOOV LPPXQHFHOOV GLIIHUHQWLDWLRQ ,QYLWUREORRGEUDLQEDUULHUPRGHO XVLQJKXPDQL36FHOOV VWHPFHOOV YDVFXODUHQGRWKHOLDOFHOOV 䞉'HYHORSPHQWRIGUXJVFUHHQLQJV\VWHP 䞉'HYHORSPHQWRILQYLWURGLVHDVHPRGHO Research topics 1) Differentiation into hematopoietic and vascular endothelial cells from iPS cells. 2) Development of drug-screening system by using iPS cell-derived cells. 3) Development of in vitro disease model by using iPS cells. Recent publications 1) Minami H. et al., Generation of brain microvascular endothelial-like cells from human induced pluripotent stem cells by co-culture with C6 glioma cells. PLOS ONE, 10(6):e0128890, 2015. 2) Tashiro K. et al., Expression of coxsackievirus and adenovirus receptor separates hematopoietic and cardiac progenitor cells in fetal liver kinase 1-expressing mesoderm. Stem Cells Translation. Med., 4, 424, 2015. 3) Tashiro K. et al., Plasma elevation of vascular endothelial growth factor leads to the reduction of mouse hematopoietic and mesenchymal stem/progenitor cells in the bone marrow. Stem Cells Dev., 23, 2202, 2014. 4) Yamaguchi T. et al., Two-step differentiation of mast cells from induced pluripotent stem cells. Stem Cells Dev., 22, 726, 2013. 35 Laboratory of Clinical and Experimental Pathophysiology Professor Masahiro INOUE +81-6-6972-1181 [email protected] Staff Hiroko ENDO +81-6-6972-1181 endou-hi@ mc.pref.osaka.jp FAX +81-6-6973-5691 Our research aims at understanding cancer biology and developing novel therapeutics and diagnostics against cancer. We are currently focusing on 1) culture of primary cancer cells and 2) caner metabolism. 1) Culture of primary cancer cells We recently developed novel methods to prepare and culture cancer cells from patient tumor samples. The methods, CTOS (cancer tissue-originated spheroids), enable easy preparation of pure cancer cells from the biopsy samples with high efficiency. Cancer cells in CTOS preserve original characters even after long time culture. CTOS culture can be applicable to the personalized medicine as well as the development of new drugs. Figure 1 A Method for preparation and culture of 3D primary cancer cells CTOS: cancer tissue-originated spheroid Tumor samples &KDUDFWHULVWLFVRI&726 Organoid Pure cancer cells High yield Dissociation Filter CTOS Stable culture Passageable Flow through Tumorigenic in mice Retaining original characters 100mm 2) Cancer metabolism Hypoxia is highly related to the malignant and refractory features of cancer. The cancer cells adapt to the hypoxic microenviron- ment, altering energy metabolism as well as enhancing anti-death machinery. We are focusing on the mechanism how the cancer cells can survive under limited energy source. We have clarified that suppression of mTOR and c-Myc are involved in the hypoxia resistance of cancer cells. Tumor cells may circumvent critical decrease of oxygen concentration in their microenvironment by suppressing their oxygen consumption. Figure 2 Hypoxic microenvironment in solid cancer 3 Adoptation 1 Renovation of microenvironment Hypoxic region (Metabolism) (Anti-death) (Angiogenesis) Oxygenized region 2 Escape (invasion, matastasis) Cancer cells Blood vessel Xenograft of human colorectal cancer Research topics 1) Culture of primary cancer cells 2) Cancer metabolism Recent publications 1) Sato Y, et al. Generation of a monoclonal antibody recognizing the CEACAM glycan structure and inhibiting adhesion using cancer tissue-originated spheroid as an antigen. Sci Rep 2016; 6: 24823. 2) Okuyama H, et al. Dynamic Change of Polarity in Primary Cultured Spheroids of Human Colorectal Adenocarcinoma and Its Role in Metastasis. Am J Pathol 2016; 186: 899-911. 3) Endo H, et al. Dormancy of Cancer Cells with Suppression of AKT Activity Contributes to Survival in Chronic Hypoxia. PLoS One 2014; 9: e98858. 4) Kondo J, et al Retaining cell-cell contact enables preparation and culture of spheroids composed of pure primary cancer cells from colorectal cancer. Proc Natl Acad Sci USA, 2011; 108:6235-40. 36 Laboratory of Biopharmaceutical Science Professor Akiko ISHII-WATABE +81-3-3700-9064 [email protected] FAX +81-3-3700-9084 Division of Biological Chemistry and Biologicals, National Institute of Health Sciences 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501 Japan Professor Akiko ISHII-WATABE Biotechnological products are the drugs manufactured using biotechnologies such as genetic recombination and cell culture technologies. Most of them, the active pharmaceutical ingredients are recombinant proteins. To date, more than 120 biotechnological products including insulins, erythropoietins, interferons, interleukins, enzymes, Fc-fusion proteins and therapeutic monoclonal antibodies (mAbs) have been approved in Japan as well as overseas. Recently development of next-generation mAbs with uniquely engineered structure and/or novel functions are increasingly important especially in the fields of cancers and immune diseases. Our laboratory focuses mainly on regulatory science research to accelerate the development of next-generation therapeutic antibodies and to ensure their quality, safety and efficacies. Since mAbs have Fc domain as their common structure, which relates to their immune effector functions and pharmacokinetics, we are making efforts to reveal the role of Fc receptor family molecules in safety and efficacy of newly developed mAbs. Human Fc receptor family is different from rodents, therefore in vitro evaluation method is important to predict the safety of newly developed mAbs. In addition, studies on immunogenicity assessment and development of analytical methods using emerging technologies for characterization of biotechnological product including mAbs are also conducted. Research topics 1) Studies on roles of FcȖ receptor familes in regulating the safety and efficacy of mAbs, and development of their evaluation methods 2) Studies on roles of neonatal Fc receptor (FcRn) in regulating the pharmacokinetics and other aspects of mAbs 3) Development of in vitro safety evaluation method of next-generation mAbs 4) Studies on immunogenicity assessment of biotechnological products 5) Development of analytical method for characterizing the biotechnological products Recent publications 1) Ishii-Watabe A. et al. Approaches to Quality Risk Management When Using Single-Use Systems in the Manufacture of Biologics. AAPS PharmSciTech 16, 993-1001 (2015) 2) Tada M. et al. Characterization of anti-CD20 monoclonal antibody produced by transgenic silkworm (Bombyx mori). mAbs 7, 1138-1150 (2015) 3) Suzuki T et al. Fluorescent imaging method for analyzing the biodistribution of therapeutic monoclonal antibodies, which can distinguish intact antibodies from breakdown products. mAbs 7, 759-769 (2015) 7DGD0HWDO'HYHORSPHQWRIDFHOOEDVHGDVVD\PHDVXULQJWKHDFWLYDWLRQRI)FȖ5,,DIRUWKHFKDUDFWHUL]DWLRQRIWKHUDSHXWLF monoclonal antibodies PLOS ONE 9, e95787 (2014) 5) Ishii-Watabe A. et al. [Nonclinical Evaluation of Next-generation Therapeutic Monoclonal Antibodies]. YAKUGAKU ZASSHI 135, 857-866 (2015) (in Japanese) 37 Laboratory of Functional Pharmaceutics Professor Haruhiro OKUDA +81-3-3700-2859 [email protected] Associate Professor Yukio ASO +81-3-3700-8547 [email protected] Pharmaceuticals are industrial products with specific characteristics： They affect life of human directly. Nevertheless, consumer cannot obtain information about their quality easily. They have long life cycles as advanced industrial products. Tests for efficacy and safety are not conducted at the release of them. Tests of efficacy and safety of a pharmaceutical are not conducted at the release, since efficacy and safety of pharmaceuticals are confirmed by producing pharmaceuticals by well-controlled manufacturing process and conducting quality tests at the release. Therefore, it is important to identify quality attributes that affect the efficacy and safety of the pharmaceuticals (critical quality attributes, CQA), and it is essential to develop a manufacturing process that produce the pharmaceuticals with consistent CQA and to control the process appropriately. Laboratory of Functional Pharmaceutics is conducting researches on the quality of pharmaceuticals with advanced functionality, especially the quality of pharmaceuticals with improved solubility characteristics, in order to assure efficacy and safety of the pharmaceuticals. Recently, lead compounds of active pharmaceutical ingredient (API) are optimized by docking study with lipophilic active site, and poorly water soluble compounds are frequently selected as candidates of API. Poor water solubility of the lead compounds becomes a significant hurdle to commercialization of the lead compounds. To improve solubility of poorly water soluble lead compounds, the high energy form of lead compounds such as amorphous form, nano-particles and co-crystal form are studied for producing drug products. Technology of high energy form of lead compounds is going to be an indispensable part for suppling innovative pharmaceuticals to medical front, and a lot of researches are conducted especially for the lead compounds of amorphous state. On the other hand, the high energy form pharmaceuticals are inherently unstable, and very small variation of process parameter may cause problem of the quality, which may fail in expected bioavailability. We are studying to clarify the factors which affect the quality variation of high energy form pharmaceuticals, and promoting research on the regulatory science about quality of high energy form pharmaceuticals. (Figure) ƌLJƐƚĂůůŝnjĂƚŝŽŶŽĨĂŵŽƌƉŚŽƵƐŶŝĨĞĚŝƉŝŶĞ ĂƚƚŚĞƐƵƌĨĂĐĞ ŽďƐĞƌǀĞĚďLJĂƚŽŵŝĐĨŽƌĐĞŵŝĐƌŽƐĐŽƉĞ;Ϯϲ㼼ϭΥͿ Research topics 1) Elucidation and control of factors affecting on the quality of high energy form pharmaceuticals 2) Study on stability prediction based on the physico-chemical property of high energy form pharmaceuticals Recent publications 1) Miyazaki T. et al., Feasibility of atomic force microscopy for determining crystal growth rates of nifedipine at the surface of amorphous solids with and without polymers. J. Pharm. Sci., 100, 4413-4420, 2011. 2) Miyazaki T. et al., Differences in crystallization rate of nitrendipine enantiomers in amorphous solid dispersions with HPMC and HPMCP. Int. J. Pharm., 407, 111-118, 2011. 3) Miyazaki T. et al., Physical characterization of dibasic calcium phosphate dehydrate and anhydrate. J. Pharm. Sci., 98, 905-916, 2009. 4) Aso Y. et al., Feasibility of 19F-NMR for assessing the molecular mobility of flufenamic acid in solid dispersions. Chem. Pharm. Bull., 57, 61-64, 2009. 38 Laboratory of Food Safety Professor Hiroshi AKIYAMA +81-3-3700-2158 [email protected] Associate Professor Hiromi NABESHI +81-3-3700-9359 [email protected] Professor Hiroshi AKIYAMA In the field of pharmaceutical science, the domains of regulatory science (RS) include pharmaceuticals, foods, and living environ- ments. For pharmaceuticals, efficacy are given priority in the balance between efficacy and safety to cure a disease, and the distri- bution to the public is judged from the balance. For foods, on the other hand, safety is the first priority to the public acceptance. To ensure food safety, hazards in foods or hazardous foods with the potential to cause adverse health effects should be identified and characterized. Therefore, bacteria and virus which cause food poisoning, natural toxin, pesticide residues, veterinary drug residues, food additives, other chemical substances, radioactive substances, and allergens in foods should be identified and charac- terized. In addition, risk assessment which assesses the degree of the risk for human health and environmental health, and risk management which takes measures to reduce the risk should be conducted. The effectiveness of the measures by risk management are monitored, and if they did not work effectively, the risk assessment and the risk management should be reviewed. These proce- dures also need risk communication, the exchange of the opinions and information on the risk among producers, consumers, and the government. Above all measures to assure food safety is called risk analysis. Risk analysis consists of risk assessment, risk management, and risk communication. Risk analysis is a procedure to adjust the risk when the public are exposed to the risk, to prevent accidents in advance as possible, and to reduce the risk to the minimum. Risk analysis and measures to food chain are both necessary for the stable supply of safe foods. RS in the field of food safety is the science which supports appropriate prediction, assessment, and judgement of food for risk analysis, in other words, the science which supports risk analysis of foods. From a viewpoint of food safety, to assure the public health is most important. However, the establishment of rules on production and distribution of foods is also essential, because the proper system of agriculture and food industries is necessary to secure the supply of foods for the public. RS in the field of food safety plays a part in establishing rules to apply them to the field of living environments through risk analysis of food safety. Our major studies are as follows. i) Development of the analytical methods on pesticide residues, veterinary drug residues, environmental pollutants, and the pollutant such as heavy metal, and the assurances of the quality ii) Investigation of dietary intake of the pollutants iii) Analysis and investigation of substances generated by food processing and radioactive substances in foods iv) Development of the detection methods of irradiated foods In studies of i), we have already developed multi-residue methods for analy- sis of agricultural chemicals using a liquid chromatography time-of-flight mass spectrometry (LC-TOF/MS). Development of simple and cyclopedic analytical methods for all chemicals is our current important mission. Our main studies are categorized into the science which mainly supports risk management and our goal is to reflect the results of studies to the national regulatory policies. While the studies on dietary intake surveys of chemical substances are to provide the scientific evidence to the risk assessment. We could say that our studies which provide scientific evidence to risk management and risk assessment are RS in the field of food safety. We conduct the studies daily for the development of evaluation methods on food safety, considering implementation of novel innovated methods and conducting collaborative studies with other research institutes. Research topics 1) Studies on development of the evaluation methods of the sensitization for food additives 2) Studies on establishment of standards for safety assurance of existing food additives 3) Studies on development of the analytical methods of pesticide residues and veterinary drugs in foods 4) Studies on evaluation of dietary intake of harmful substances such as dioxins via food and development of the analytical methods 5) Studies on the analysis and investigation of radioactive substances and harmful chemical substances caused by the Great East Japan Earthquake Recent publications 1) Akiyama H., Matsuoka H., et al, The acute encephalopathy induced by intake of Sugihiratake mushroom in the patients with renal damage might be associated with the intoxication of cyanide and thiocyanate, Food Safety, 3, 16-25, 2015. 2) Koizumi D, Akiyama H, et al, Development and validation of a lateral flow assay for the detection of crustacean protein in processed foods, Food Chemistry, 150, 348-352, 2014. 3) Saito S., Nemoto S., Matsuda R., Multi-residue analysis of pesticides in agricultural products by liquid chromatography time- of flight mass spectrometry. Food Hyg. Saf. Sci., 53, 255-263, 2012. 4) Kojima H., Takeuchi S., Tsutsumi T. et al., Determination of dioxin concentrations in fish and seafood samples using a highly sensitive reporter cell line, DR-EcoScreen cells, Chemosphere, 83, 753-759, 2011. 5) Watanabe T., Maitani T., Matsuda R., Analysis of the distribution of analyte concentration in lot, sample size, and number of analytical runs on food-testing results. J. Agric.Food Chem, 60, 10702-10708, 2012. 39 Laboratory of Pharmaceutical and Food Microbiology Associate Professor Yutaka KIKUCHI +81-3-3700-9496 [email protected] (Lead Scientist) FAX +81-3-3700-9496 Associate Professor Yutaka KIKUCHI The research interests are focused on microbiological safety in the pharmaceutical filed. Transmissible spongiform encephalopathy (TSE) is a neurodegenerative disease characterized by spongiform degeneration and accumulation of an infectious isoform (PrPSc) of the prion protein in the central nervous system. Some studies reported the possible case of variant Creutzfeldt-Jakob disease (vCJD), human prion disease originates from bovine spongiform encephalopathy (BSE), acquired by blood transfusion. Another route of transmission of vCJD could be via cell substrates using contaminated blood or serum. We attempted to develop a method of quantitative analysis of PrPSc that are used in the manufacture of biotechnological/biological products. In addition, we interested in physical role of prion protein. The human prion protein (PrP) is a glycoprotein with a glycosylphosphatidylinositol (GPI) anchor at its C-terminus. We report alternative splicing within exon 2 of the PrP gene (PRNP) in the human glioblastoma cell line T98G. An open reading frame of the alternatively spliced mRNA lacked the GPI anchor signal sequence and encoded a 230-amino acid polypeptide. Its product, GPI-anchorless PrP (GPI¯PrPSV), was unglycosylated and nonionic detergent-soluble and was found in the cytosolic fraction. We also detected low levels of alternatively spliced mRNA in human brain and non-neuronal tissues. When long-term passaged T98G cells were placed in a low-oxygen environment, alternatively spliced mRNA expression increased while ordinary PrP mRNA expression decreased. These findings imply that oxygen tensions regulate GPI¯PrPSV expression in T98G cells. We have been also studied about the methods for ensuring microbiological safety including bacterial endotoxin test, sterility test and mycoplasma test for biotechnological, biological and regenerative medicine products. Furthermore, we focus on the pathogenic mechanisms of unidentified foodborne disease associated with fresh food. Therefore, specific antibodies against causative agents of unidentified foodborne disease have been developed that would support the identification of pathogens. Research topics 1) Studies on the quantitative methods for analysis of PrPSc that are used in the manufacture of biotechnological/biological products. 2) Studies on the methods for ensuring microbiological safety of biotechnological, biological and regenerative medicine products. 3) Immunochemical detection of causative agents of unidentified foodborne disease associated with fresh food. Recent publications 1) Kanayasu-Toyoda, T. et al.: Cell-Surface MMP-9 Protein Is a Novel Functional Marker to Identify and Separate Proangio- genic Cells from Early Endothelial Progenitor Cells Derived from CD133(+) Cells. Stem Cells, 34, 1251-1262, 2016. 2) Watanabe, M., Kikuchi, Y.: Study on the Detection and Identification Method of the Fungal Test Based on Nucleic Acid Amplification Test (NAT). Pharmaceutical and Medical Device Regulatory Science, 47, 150-157, 2016. 3) Uchida, E. et al.: Collaborative Study on the PCR Detection Method of Mycoplasma Testing for Cell Substrates; The Revision for General Information of JP. Pharmaceutical and Medical Device Regulatory Science, 45, 442-451, 2014. 4) Kikuchi, Y., et al.: ELISA Detection of Kudoa septempunctata in Raw Paralichthys olivaceus (Olive Flounder) using a Chicken Anti-Kudoa Antiserum. Biocontrol Science, 18, 193-197, 2013. 5) Ohnishi, T., et al.: Kudoa septempunctata Invasion Increases the Permeability of Human Intestinal Epithelial Monolayer. Foodborne Pathog Dis., 10, 137-142, 2013. 6) Yusa, S., et al.: Cellular prion protein: from physiology to pathology. Viruses, 4, 3109-3131, 2012. 40 Laboratory of Medical Device Safety Professor Yuji HAISHIMA +81-3-3700-4842 [email protected] FAX +81-3-3707-6950 Professor Yuji HAISHIMA For development and industrialization of medical devices, there are many tasks for clearing a hurdle in the needs grasp of the healthcare field or the correspondence to a pharmaceutical approval. In addition, the problems such as reclamation or the standard- ization strategy of the market pile up after marketing exceedingly. As one of national strategy to break off these problems, act to advance health and medicine strategy was established, and the promotion of “Regulatory Science” for properly predicting and evaluating the quality, effectiveness, and safety of medical devices based on scientific basis is being exhorted to occasion of practi- cal use as the results of research and development in medical field. Material selection for the development of medical devices with innovative technology is important for exhibiting the intended function, and novel testing methods for evaluating the safety and effectiveness are often required. It becomes possible to distribute a safer medical device to patients by developing the methods according to the characteristic of the products. Moreover, the existing methods also need to be revised by the content corresponding to the present science level. Thus, the importance of a nonclinical test is now increasing for the safety assessment of medical devices applied to human body, and hence we powerfully promote the study on a development and standardization of the new examining method. In addition, as a part for accelerating the life cycle of medical devices, we are promoting the preparation and standardization of a standard, study on development of novel medical devices and biomaterials, participation to support network for development of medical devices, human resource training, and public-relations / public awareness campaign under the cooperation with industry, government, and the local community including a lot of university. The content that should be shown in a clinical trial is often decided according to the results yielded in a nonclinical study, and the items that should be observed in a clinical trial will be decided if a hazard and risk is fully detected and presumed by a nonclinical test. Thus, a nonclinical test is very important for considering the design of a clinical trial. We have mainly developed and improved horizontal testing methods for medical devices and biomaterials, but these general methods are often inapplicable for the evaluation of medical devices with innovative technology. Therefore, performance test methods that can contribute to design of clinical trial and evaluate the parameters non-detectable in animal experiment also need to develop for each product group. We are also promoting the development and improvement of medical devices and biomaterials on the basis of knowledge yielded in the safety evaluation study. Research topics 1) Development of chemical, physical, biological, microbial and mechanical safety evaluation method of medical devices. 2) Development of novel blood containers and circuits made of polyvinyl chloride with alternative plasticizers. 3) Development of novel sterilizer with the mixed gas of ozone and hydrogen peroxide. 4) Development of novel functional biomaterials using RNA aptamer. 5) Development of surgical navigation system for real-time safe and secure surgery. Recent publications 1) Haishima Y, et al., Endotoxin limit of biomaterials for bone regeneration in cranial and femoral implantation of rats. J. Biomed. Mater. Res. Part B. in press, 2016. 2) Haishima Y, et al., Characterization of alternative plasticizers in polyvinyl chloride sheets for blood containers. J. Vinyl Add. Technol. in press, 2016. 3) Haishima Y, et al., Screening study on hemolysis suppression effect of an alternative plasticizer for the development of a novel blood container made of polyvinyl chloride. J. Biomed. Mater. Res. Part B. 102, 721-728, 2014. 4) Haishima Y, et al., Development and performance evaluation of a positive reference material for hemolysis testing. J. Biomed. Mater. Res. Part B. 102, 1809-1816, 2014. 41 Project of Clinical Pharmacy Research and Education Professor Etsuko UEJIMA +81-6-6879-8251 [email protected] Associate Professor Kazuyuki NIKI +81-6-6879-8250 [email protected] Specially Appointed Assistant Professor Asuka HATABU +81-6-6879-8250 [email protected] Advanced Project of Clinical Pharmacology Professor Yasushi FUJIO +81-6-6879-8258 [email protected] Clinical pharmacology is the research field that addresses and clinical profiles, including backgrounds of the patients, PK pharmaco-kinetic and -dynamic properties of drugs in human parameters and clinical outcomes. The goal of this project is to in order to design sophisticated pharmacotherapy based on establish individualized medicine of anti-cancer drug therapies. scientific data. The aim of this project is to develop the human [Project 2] PGx studies for smoking cessation. resources that contribute to optimization of drug therapies Smoking is considered an intractable disease designated as though clinical pharmacology. Two projects are now on-going. nicotine dependence. We previously reported that nicotine [Project 1] Clinical pharmacogenomics (PGx) studies for dependence is closely associated with the genetic polymor- pharmacokinetics (PK)-related genes. Though much infor- phisms of CYP2A6, a metabolizing enzyme of nicotine, and mation upon genetic polymorphisms of PK-related genes has that the patients with genotypes of high CYP2A6 activities been accumulated, genetic information remains to be fully exhibit serious nicotine dependence. In this project, we are utilized in medical examination and treatment in practice. In planning to perform genome-wide association study (GWAS) this project, we focus on anti-cancer drugs. We analyze the to explore the genetic variants that are associated with nicotine relation between genetic polymorphisms of PK-related genes dependence or adverse events of anti-smoking drugs. Recent publications 1) Nakagawa, M. et al. Four cases of investigational therapy with interleukin-11 against acute myocardial infarction. Heart and Vessels. 2016; 31: 1574-1578. 2) Takimoto, T., et al. Polymorphisms of CYP2D6 gene and gefitinib-induced hepatotoxicty. Clin. Lung Cancer. 2013; 14:502-7. 3) Azuma, J. et al. NAT2 genotype guided regimen reduces isoniazid-induced liver injury and early treatment failure in the 6-month four-drug standard treatment of tuberculosis: A randomized controlled trial for pharmacogenetics-based therapy. Eur. J. Clin. Pharmacol. 2013; 69, 1091-101. 4) Yamada, T. et al. Genetic polymorphisms of CYP17A1 in steroidogenesis pathway are associated with risk of progression to castration-resistant prostate cancer in Japanese men receiving androgen deprivation therapy. Int. J. Clin. Oncol. 2013; 18, 711-7. 5) Kijima, T., et al. Safe and successful treatment with erlotinib after gefitinib-induced hepatotoxicity: difference in metabolism as a possible mechanism. J. Clin. Oncol. 2011; 29, e588-90. 42 Education and Research Project of Pharm.D/ Laboratory of Advanced Research of Medical and Pharmaceutical Sciences Professor (coordinator) Tadayuki UNO +81-6-6879-8205 [email protected] Professor (coordinator) Yasushi FUJIO +81-6-6879-8258 [email protected] Associate Professor Shinichiro MAEDA +81-6-6879-3372 [email protected] FAX +81-6-6879-3379 Education and Research Project of Pharm.D was newly established in 2013, and focused on propulsive the educational and research curriculum of Pharm.D (Osaka University), first established in Japan. Pharm.D course had started in USA from the latter part of the twentieth century; specialized in clinical pharmacy education curriculum. Our project aimed to producing quality graduates of clinical pharmacists, project manager of translational research, and public administrative officer of pharmaceutical affairs. To these ends, various practical training was designed; such as, community pharmacy, hospital pharmacy, acute phase hospital ward, medical center for translational research, lectures of clinical investiga- tion and medical statistics, public administration internship, and overseas training. Laboratory of Advanced Research of Medical and Pharmaceutical Sciences was focused on research of clinical pharmacy discipline, such as pharmacokinetics, clinical pharmacology, and drug-drug interaction. Research topics 1) Therapeutic drug monitoring (TDM) of anticancer agent, immune-suppressive agent. 2) Pharmacokinetics study of anticancer agent, immune-suppressive agent. 3) Clinical genome pharmacology study of drug-metabolizing enzyme system 4) Drug-drug interaction study of immune-suppressive agent. Recent publications 1) Maeda S. et al., Multicomponent high-performance liquid chromatography/tandem mass spectrometry analysis of ten chemo- therapeutic drugs in wipe samples. J Chromatogr B Analyt Technol Biomed Life Sci 921-922, 43-48 (2013). ஜന૯֚ͣȂࢯ̦ͭसईࣣୋުྩ̤̫ͥͅհ͈બȡ10 ਅ႒͈सͬచય̱̹͂ࢯ̦ͭसईࣣୋশ (2 .(அऔȡ 48 (10), 2415-2421 (2012ޏ۪̫̤ͥͅ 3) Maeda S. et al., Environmental Contaminations and Occupational Exposures Involved in Preparation of Chemotherapeutic Drugs. Environmental Contamination (InTech), chapter 5, 81-92 (2012). 4) Maeda, S., et al., Evaluation of environmental contaminations and occupational exposures involved in preparation of chemo- therapeutic drugs. Yakugaku Zasshi 130 (6), 903-910 (2010). Project for Integrative Education in Pharmaceutical Sciences and Regional Medical Treatment Professor Kazumasa HIRATA +81-6-6879-8236 [email protected] Specially Appointed Assistant Professor Misa MURAOKA +81-6-6879-8236 [email protected] Project for Integrative Education in Pharmaceutical Sciences and Regional Medical Treatment is a division of Center for Advanced Education and Research in Pharmaceutical Sciences founded in 2005. This project aims to clarify the responsibility for planning and management of education programs in conformity with “Education Philosophy and Objective” of the School and Graduate School of Pharmaceutical Sciences. This project develops and manages the education projects promoted by Ministry of Education, Culture, Sports, Science and Technology under collaboration with other universities and medical organizations like hospitals and pharmacy in Osaka Prefecture and Kinki Region. Research topics 1) Collaborative Development of Advanced Practical Education Program to Train and Support Pharmacists with Prominent- Leadership (2016-2021) 2) Problem-solving Oriented Training Program for Advanced Medical Personnel (2014-2018) Recent publications 1) Hirata, K. et al. Collaborative development of advanced practical education program to train pharmacists with leadership under the joint operation of pharmaceutical departments of fourteen national universities. Yakugaku Zasshi, 132, 365-368 (2012). 2) Takiguchi Y., Hirata, K. et al. Development of educational evaluation methods in practical experience in national universities. 132, 365-368 (2012). 43 Platform Project for Drug Discovery Professor Tadayuki UNO +81-6-6879-8205 [email protected] Assistant Professor Hirofumi TSUJINO +81-6-6879-8208 [email protected] FAX +81-6-6879-8209 Professor Tadayuki UNO Our platform project for drug discovery is financially supported by MEXT, and plays a key role in drug discovery and develop- ment in Osaka University. This project is aimed at construction of an innovative process for drug discovery with our high- throughput screening facilities, supporting academic researchers who have highly original drug seeds. In addition, we construct a research and education system, cooperating with research institutes and pharmaceutical companies in Japan, to train young researchers who will contribute to innovative drug development in the future. Unique research projects are now going on, and we welcome students who are interested in drug discovery and development in academia. Research topics 1) Development of a drug which inhibits immune escape of cancer cells 2) Construction of a high-throughput assay system for ldrug metabolism Recent publications 1) Miyamoto M. et al., Membrane anchor of cytochrome P450 reductase suppresses the uncoupling of cytochrome P450. Chem. Pharm. Bull. 63, 286-294, 2015 2) Tsujino H. et al., Disulfide Bonds Regulate Binding of Exogenous Ligand to Human Cytoglobin. J. Inorg. Biochem. 135, 20-27, 2014 Lead Exploration Units Professor Kazutake TSUJIKAWA +81-6-6879-8190 [email protected] Professor Shuji AKAI +81-6-6879-8210 [email protected] Specially Appointed Professor Jun-ichi HARUTA +81-6-6879-5779 [email protected] Guest Professor Ryu NAGATA +81-6-6879-5781 [email protected] Specially Appointed Associate Professor Masahiro SAKAGAMI +81-6-6879-5781 [email protected] Specially Appointed Associate Professor Iichiro KAWAHARA +81-6-6879-5781 [email protected] Specially Appointed Associate Professor Koji MATSUMURA +81-6-6879-5781 [email protected] Specially Appointed Associate Professor Kozo YOSHIDA +81-6-6879-5781 [email protected] Specially Appointed Associate Professor Hiroshi NAKAGAWA +81-6-6879-5781 [email protected] Specially Appointed Assistant Professor Shinsuke KOMAGAWA +81-6-6879-5781 [email protected] Specially Appointed Assistant Professor Masayuki AZUMA +81-6-6879-5781 [email protected] Specially Appointed Assistant Professor Shohei SATO +81-6-6879-5781 [email protected] Specially Appointed Assistant Professor Shintaro FUJII +81-6-6879-5781 [email protected] Specially Appointed Researcher Kazuto NUNOMURA +81-6-6879-5781 [email protected] Specially Appointed Researcher Bangzhong LIN +81-6-6879-5781 [email protected] The goal in this Units is to create clinical candidate low-molecular compounds for treating intractable and rare diseases based on biological research outcomes at academia. The leading-edge sciences such as chemistry, biology, medicine and physics are extremely important for the research of drug discovery without doubt. Specifically, we are engaged in drug design and synthesis of compounds by experts in medicinal chemistry, and drug metabolism and pharmacokinetics (DMPK) through the optimization of lead compounds. In this way, we make an effort to get candidates having good balance among biological activity, physical property, DMPK and toxicity. After that, the candidates obtained in this Units will be transferred to pharmaceutical companies from academia. Finally, this Units shall also educate young researchers who will lead medicinal chemistry. 44 Structural Biology Project Associate Professor Hiroshi AOYAMA +81-6-6879-4095 [email protected] X-ray crystallography is the best technique for determination of three-dimensional structure of molecules, such as organic molecules, DNA duplexs and proteins. The main aim of our research is X-ray structure determination of biological macromolecular assemblies including membrane protein complexes. In addition to the understanding of the relationship of structure and function, our aim is the application and development of structures, such as drug discovery. To achieve these aims, we employ a varied experimental approach that includes molecular biology, analytical chemistry, physical chemistry and computer science. We welcome motivated students who are interested in our Crystal structure of cytochrome c oxidase research field. (PDB ID : 3ABM, 2009) Research topics 1) X-ray crystallography of mitochondrial cytochrome c oxidase, other proteins, organic molecules and DNA duplexs 2) Structure and mechanism of cell-cell interactions Recent publications 1) Bai D. et al., Engineered Cx26 variants established functional heterotypic Cx26/Cx43 and Cx26/Cx40 gap junction channels Biochem J., 473, 1391-1403, 2016 2) Murai K. et al., Oxidative rearrangement via in situ generated N-chloroamine : Synthesis of fused tetrahydroisoquinolines Org. Lett., 18, 1314-1317, 2016 3) Takashima S. et al., Higd1a is a positive regulator of cytochrome c oxidase Proc. Natl. Acad. Sci. USA, 112, 1553-1558 2015 Unit for Pharmaceutical Education in Community Medical Care Professor Kazumasa HIRATA +81-6-6879-8236 [email protected] Specially Appointed Professor Takao NISHINO +81-6-6879-8239 [email protected] Specially Appointed Assistant Professor Misa MURAOKA +81-6-6879-8236 [email protected] Unit for Pharmaceutical Education in Community Medical Care is a division of Center for Pharmaceutical Education and Research in Community Medical Care founded in 2015. This unit aims to clarify the responsibility for planning and management of education programs in conformity with “Education Philosophy and Objective” of the School and Graduate School of Pharmaceutical Sciences. The objectives of this unit include development of advanced education and research programs, management of pharmacist-training programs in hospital and pharmacy, planning of Faculty Development programs, and management of the education projects promoted by Ministry of Education, Culture, Sports, Science and Technology. Research topics 1) Collaborative Development of Advanced Practical Education Program to Train and Support Pharmacists with Prominent- Leadership (2016-2021) 2) Problem-solving Oriented Training Program for Advanced Medical Personnel (2014-2018) Recent publications 1) Hirata, K. et al. Collaborative development of advanced practical education program to train pharmacisits with leadership under the joint operation of pharmaceutical departments of fourteen national universities. Yakugaku Zasshi, 132, 365-368 (2012). 2) Takiguchi Y., Hirata, K. et al. Development of educational evaluation methods in practical experience in national universities. 132, 365-368 (2012). 45