ポスター
プログラムおよび要旨集
Welcome
Address
Dean Prof. Yoshinori
Moriyama
Faculty of Pharmaceutical Sciences,
Okayama University
First of all, I would like to express my sincere thanks to the members
of Sungkyunkwan University in South Korea who visited us at Okayama University.
When we visited School of Pharmacy, Sungkyunkwan University, in this September,
I found an agreement form which was exchanged more than 10 years ago on
display in a safe place in the dean’s room. I was very pleased to
learn that the faculty members of Sungkyunkwan University consider
us as an important partner. Now, we have opportunity to show off
our research to each other at Okayama University. I hope that
all of participants enjoy this symposium, providing important progress
for mutual understanding, collaboration and the development of both faculties.
1
(OU-1)
Molecular imaging technology on the
research of pharmaceutical sciences
Shuichi
Enomoto1,2,3
1 Professor, Department of Pharmaceutical Analytical Chemistry, Graduate
School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University,
Okayama, Japan,
2 Professor, Okayama Medical Innovation Center of Molecular Imaging,
Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama
University, Okayama, Japan,
3 Head, Next-generation Imaging Team, Center for Life Science Technologies, RIKEN Kobe
Institute, Japan
Rapidly occurring discoveries and innovations in molecular biology, imaging
and computing technology and driving a new scientific specialty referred
to as "molecular imaging". Although mmolecular imaging is broadly
defined as the characterization and measurement of biological processes
in living animals, model systems and humans at the cellular and molecular
level, using remote imaging detection methods, such as PET, SPECT, MRI
and optical imaging and is helping us to better understand many of the
basic molecular mechanisms of disease, the ability to use imaging to drive
"personalized medicine" in patients is still a distant goal.
Translational imaging research will facilitate the goal. Our mission is
to: a) Translate advances in cellular and molecular biology, chemistry,
physics, computer sciences, engineering, instrumentation and animal models
into improvements in care for patients, b) Provide further advances in
understanding of the molecular basis of disease and, c) Train student on
undergraduate or graduate school and investigators in the use of modern
molecular imaging techniques. The focus of our laboratory is the
dynamics of bio-molecules on living organisms, as measured using various
molecular imaging modalities, PET, SPECT, CT, MRI, Optical imaging technologies
and our newly developed modality “Gamma-ray Emission Imaging (GREI)” which development of RIKEN’s original
semiconductor Compton camera GREI, and the world’s first demonstration of multiple
molecular imaging. Recently we have successfully developed several new
anti-body- and peptide-based probes for diagnostic imaging of caner,
inflammation, Alzheimer disease and diabetes. And we also have successfully
developed aminobenzopyrano-xanthene (ABPX) dyes that exhibits fluorescence
emission in both dilute solution and the aggregates.
Our recent topics
1.
T. Miyamoto, S. Kamino, A.
Odani, M. Hiromura, S. Enomoto, Basicity of N-terminal
amine in TCUN peptide regulates stability constant of albumin-like Cu2+
complex.
Chem. Lett., 42,
1099-1101 (2013).
2.
Y. Shirasaki, S. Kamino, M. Tanioka, K.
Watanabe, Y. Takeuchi, S. Komeda, and S. Enomoto, New
aminobenzopyranoxanthene-based colorimetric sensor for Cu2+ with
dual-color signal detection system., Chem.
Asian J., 8, 2609-2613 (2013).
3.
S. Motomura, Y. Kanayama, M. Hiromura, T.
Fukuchi, T. Ida, H. Haba, Y. Watanabe, and S. Enomoto, Improved imaging
performance of semiconductor Compton camera GREI makes for a new methodology to
integrate bio-metal analysis and molecular imaging technology in living
organisms., J. Anal. At. Spectrom. , 28, 934-939(2013).
4.
S. Kadowaki, M. Munekane, Y. Kitamura,
M. Hiromura, S. Kamino, Y. Yoshikawa, H. Saji, S. Enomoto., Development
of new zinc dithiosemicarbazone complex for use as oral antidiabetic agent , Biol. Trace Elem. Res., 154, 111-119 (2013).
5.
M. Taniguchi, A. Fukunaka, M. Hagihara,
K. Watanabe, S. Kamino, T. Kambe, S. Enomoto, M. Hiromura, Essential
Role of the Zinc Transporter ZIP9/SLC39A9 in Regulating the Activations of Akt
and Erk in B-Cell Receptor Signaling Pathway in DT40 Cells, PLoSOne, vol. 8; 3 e58022. (2013).
6.
S. Kamino, A. Muranaka, M. Murakami, A. Tatsumi,
N. Nagaoka, Y. Shirasaki, K. Watanabe, K. Yoshida, J. Horigome, S. Komeda, M.
Uchiyama, S. Enomoto, A Red-Emissive aminobenzopyrano-xanthene
Dye: Elucidation of Fluorescence Emission Mechanism in Solution and Aggregate
State, Phys. Chem. Chem. Phys., 15, 2131-2140
(2013).
7.
S.
Takeda, H. Odaka, S. Ishikawa, S. Watanabe, H. Aono, T. Takahashi, Y. Kanayama,
M. Hiromura and S. Enomoto, Demonstration of in-vivo multi-probe tracker based on a Si/CdTe semiconductor
Compton camera, IEEE Trans. Nucl. Sci.,
59, 70-76 (2012).
8.
K. Higashikawa,
N. Akada, K. Yagi, K. Watanabe, S. Kamino, Y. Kanayama, M. Hiromura and S.
Enomoto, Exploration of target molecules for molecular imaging of
inflammatory bowel disease, Biochem.
Biophys. Res. Commun., 410, 416-421 (2011).
9.
H.
Fujishiro, M. Doi, S. Enomoto and S. Himeno, High sensitivity of RBL-2H3
cells to cadmium and manganese: an implication of the role of ZIP8, Metallomics, 3, 710-718 (2011).
10. A. Fukunaka, Y. Kurokawa, F. Teranishi, I.
Sekler, K. Oda, M. L. Ackland, V. Faundez, M. Hiromura, S. Masuda, M. Nagao, S. Enomoto and T. Kambe, Tissue
nonspecific alkaline phosphatase is activated via a two-step mechanism by zinc
transport complexes in the early secretory pathway, J. Biol. Chem., 286, 16363-16373 (2011).
11. T. Fukuchi, Y. Arai, F. Watanabe, S.
Motomura, S. Takeda, Y. Kanayama, H. Haba, Y. Watanabe and S. Enomoto, A
digital signal processing module for Ge semiconductor detectors, IEEE Trans. Nucl. Sci., 58, 461-467
(2011).
12. M. Kidera, Y. Seto, K. Takahashi, S.
Enomoto, S. Kishi, M. Makita, T. Nagamatsu, T. Tanaka and M. Toda, New
method for comprehensive detection of chemical warfare agents using an
electron-cyclotron-resonance ion-source mass spectrometer, Spectroc. Acta Pt. A-Molec. Biomolec. Spectr., 78, 1215-1219 (2011).
13. H. Fujishiro, K. Kubota, D. Inoue, A. Inoue,
T. Yanagiya, S. Enomoto and S. Himeno, Cross-resistance of
cadmium-resistant cells to manganese is associated with reduced accumulation of
both cadmium and manganese, Toxicology,
280, 118-125 (2011).
Name,
Ph.D. Shuichi Enomoto
(Email: senomoto@ pharm.okayama-u.ac.jp)
Research
Interests
Non-natural amino acids, Epigenetic drug discovery,
Molecular design, Fragment-based
drug discovery, In-silico screening for drug candidates, ADMET (absorption,
distribution, metabolism, excretion and toxicity, Single cell analysis,
Intracellular network, Discrimination between normal cells and abnormal cells,
Data analysis, Data base integration, Nanofabrication and nanodevice, Labeling chemistry, Analysis of metabolite dynamics, Clinical test/clinical trials, Comparative
study of the normal and disease-model animals, Comparative study of the
human and nonhuman-primates, Normal stem cells vs cancer stem cells, Central
Nervous System Disorder, Regenerative medicine, Microdosing and exploratory
clinical study, Pharmacokinetics, Efficacy evaluation, Biomarkers of pathophysiology,
Molecular imaging, Multiple molecular imaging, Development of quantitative
diagnosis of presbyopia, PET, SPECT, MRI, Optical imaging, Metallomics.
2
(SKKU-1)
Recent Progress in C−H Activation and C−C Bond Formation
In Su Kim, Associate
Professor
School of
Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea
C−H bond functionalization has been
a longstanding goal in organic synthesis since it obviates the
prefunctionalization of substrates. The combination of transition metal and
directing group is a useful strategy to facilitate C−H bond cleavage, which affords
valuable transformations of C−H bond to C−C, C−X, C−O and C−N bonds.
Since the pioneering discovery of C–H bond activation by Murai, recent progress has been focused
on dehydrogenative cross-coupling between sp2 or sp3 C–H bonds and sp2 C–H bonds of arenes or alkenes. In this area
of research, various directing groups such as ketones, carboxylic acids, amides,
pyrrole/pyridine, anilides, carbamate, urea and azine-N-oxide can provide an anchor for catalytic ortho-metalation of aromatic rings. Although the reactions using arenes or alkenes as coupling partners have been
well documented, the reactions between the aromatic C–H bonds and aldehydes or α-keto acids remain relatively unexplored. From the
synthetic point of view, the direct and catalytic introduction of carbonyl
functional group into aromatic compounds via C−H bond cleavage is among the
greatest challenges in synthetic chemistry. This new method is complementary to
Friedel-Crafts acylation and directed lithiation/acylation process.
Recently, our groups reported the rhodium or palladium-catalyzed
oxidative acylation of benzamides, N-benzyltriflamides, oximes,
phenoxypyrdines and hydrazones with aldehydes or alcohols to afford the corresponding aryl ketones or 3-hydroxyisoindolin-1-ones, which are important structural
units and synthetic intermediates in pharmaceuticals, natural products, and
functional materials. Also, we found the palladium-catalyzed
decarboxylative acylation of phenylacetamides, O-phenylcarbamtes and oximes using α-keto acids in the presence of ammonium
persulfate as a convenient oxidant affording aryl ketones in moderate to good
yields.
In this
presentation, we describe our recent achievements about the development of
transition metal-catalyzed new C-C bond formation reactions including oxidative
acylation, decarboxylative acylation, tandem indole synthesis, C-2 allylation
of indoles, and etc.
In Su Kim, Ph.D.
(Email:
insukim@skku.edu)
Associate
Professor (School of
Pharmacy)
Research
Interests
C−H Activation, C−C Bond Formation, oxidative acylation,
decarboxylative
acylation, catalytic ortho-metalation
3 (OU-2)
Toward molecular
understanding of sensory systems by structural biology
Structural
Biology
[Introduction]
Sense,
such as vision, taste sensation, olfaction, audition, and touch sensation, is
an important function for living organisms to receive, transduce, integrate,
recognize and process the environmental information. For
the first step of a molecular-level approach to the question of how we
recognize environments, our lab are addressing the structure biology of sensory
receptor proteins at the front line toward the environment, which recognize
chemical or mechanical stimuli and respond to them. We are currently performing
the structural and functional analyses of taste receptors, as representatives
of chemoreceptors, and TRP channels, as representatives of mechanoreceptors.
[Methods]
The pivot of our study is structural analyses of the
proteins by X-ray crystallography. However, our target proteins are eukaryotic
membrane proteins, which are ones of the most difficult targets in the field of
structural biology. In order to achieve structural analyses of these targets,
we simultaneously carry out research and development of methods to facilitate
sample preparation, a bottleneck for membrane protein crystallography. Our main
strategies for this is utilizing the GFP-fusion technologies, recently applied
in the field of structural biology for sample evaluation and screening.
[Results
and Discussion]
Sample
preparation of our target proteins, taste receptors and TRP channels, has so
far been difficult even for the partial regions of the receptors, because of
the poor expression level and protein unstability. By making use of the
GFP-fusion strategies, we have succeeded the protein preparation of functional
regions of these receptors, and structural and functional analyses of them are
currently in progress. At this joint symposium, I will present our recent
achievement about the analyses of the regulatory region of a fungus TRP channel
(Ihara et al. J. Biol. Chem., 288, 15303, 2013), providing clues to understand the mechanism of multimodal
responses of TRP channels.
Atsuko Yamashita, Ph.D. (Email: a_yama@ pharm.okayama-u.ac.jp)
Professor (Structural Biology)
Research
Interests
Membrane Proteins, Sensory Receptors, Transporters, Ion Channels, Taste,
Mechanosensation
4
(SKKU-2)
Structure-based Virtual Screening of Chemical Database to Discover Novel Ligands
Hyun-Ju
Park, Professor
School of Pharmacy, Sungkyunkwan University
Virtual screening is generally recognized as a
valuable tool to reduce the size of a chemical library containing
a huge number of compounds to a target-focused compound library. To overcome the major bottlenecks in this area is to find
suitable method for database filtering and scoring methods for docking. Using
various available computational programs, we setup a reliable strategy for
virtual screening and applied it to the identification of small molecule hit ligands for the targets of interest.
Promising results obtained from our study will be introduced. We identified
ligands for histone deacetylase, one of epigenetic regulators; RNA secondary structures involved in -1 ribosomal frameshifting of
many viruses; and G-quadruplex DNA
structure in the
c-Myc
oncogenic promoter.
Hyun-Ju Park, Ph.D. (Email: hyunju85@
skku.edu)
School of Pharmacy, Sungkyunkwan University
Research
Interests
structure-based virtual screening, high-content fluorescence-based screening, QSAR/QSPR modeling
5 (OU-3)
Network dysfunction by
Amyloid β
oligomers
Tsuyoshi Inoue, Associate Professor
Biophysical
Chemistry
[Introduction]
Alzheimer
disease is a common form of dementia, characterized by memory impairment.
Amyloid β (Aβ), a peptide of 42 amino acids, is the cause of
Alzheimer disease. Although Aβ peptides are highly aggregated in the brain of
Alzheimer disease, recent studies have shown that a low aggregated form of Aβ,
called Aβ
oligomers, is toxic to the brain. Actually, direct injection
of Aβ
oligomers into the mouse brain induces memory impairment (Cleary et al, Nat
Neurosci, 2005), and impairs long-term potentiation in the hippocampus, a
synaptic process of memory (Walsh et al, Nature, 2002). However, it is still unknown
how neural activities are impaired by Aβ oligomers.
[Methods]
To address this question, we used electrophysiological techniques in
awake mice. Under anesthesia, a recording electrode was implanted into the
hippocampus, and an injection cannula was implanted into the lateral ventricle.
Several days later, neural activities in the hippocampus were recorded in an
awake condition, Aβ oligomers
were injected into the lateral ventricle, and changes in the hippocampal
activities by Aβ oligomers were observed. To examine learning ability in mice, we used
a novel object recognition test as a behavioral test.
[Results
and Discussion]
Theta oscillations (5-8 Hz) are characteristic
electrical activities in the hippocampus, which are important for memory
acquisition. We found that the theta oscillations were reduced by direct
injection of Aβ oligomers. We further found a peptide, which
could recover the reduced theta oscillations by Aβ oligomers. Interestingly,
learning ability in mice was impaired by Aβ oligomers, and the memory impairment
was also recovered by the peptide. Our results indicate that Aβ oligomers
reduce theta oscillations in the hippocampus, and also that the network
dysfunction and the memory impairment by Aβ oligomers can be both recovered by
the peptide we found.
Tsuyoshi Inoue, Ph.D. (Email: tinoue@pharm.okayama-u.ac.jp)
Associate Professor (Biophysical Chemistry)
Research Interests
Electrophysiology, Neuroscience, Alzheimer disease
6 (SKKU-3)
Host-pathogen
interaction during host invasion of pneumococcus
Dong-Kwon Rhee, Professor
Molecular Microbiology Lab
[Overview]
Streptococcus
pneumoniae (pneumococcus) is the
major cause of community-associated pneumonia, otitis media, septicemia, and meningitis. Pneumonia has
one of the highest morbidity and mortality rates
from infections (≥ 2 million deaths every year), and is the
sixth-leading cause of death in the United States. It is
also responsible for a large number of deaths following influenza epidemics.
Pneumococci produce a thick polysaccharide capsule that shields pneumococci
from host phagocytes, however this
capsule is removed during invasion into the host cells. Moreover, the host
cells respond rapidly to the invading pathogens to avoid or neutralize
invasion of the pneumococcus. Thus understanding of pathogen-host interaction could provide information on chemotherapeutic measures useful
for prevention and treatment of pneumococcal
diseases.
Our lab found that 1) Pep27, an autolysis-inducing factor of S. pneumoniae, was thus expected to effect cytotoxicity. The loss
of Pep27 had a much larger than expected decrease in toxicity and has made the pep27 mutant strain sufficiently
non-toxic to be used as a live vaccine. 2)
ClpL, a
major heat shock protein (HSP) in pneumococci, generates antibiotic resistance;
Penicillin resistance in S. pneumoniae
was induced by ClpL. A mutant lacking ClpL was more susceptible to
penicillin and had a thinner cell wall than
the parental type, whereas a ClpL overexpressing strain
shows a higher
resistance to penicillin and a
thicker cell wall in the wild type. Heat shock induced a
ClpL-dependent increase in the mRNA levels and
protein synthesized by the major cell wall synthesis gene pbp2x. Fractionation
and electron micrograph data revealed that ClpL induced by heat shock is localized at the cell wall, and the DclpL
showed significantly reduced net translocation
of PBP2x into the
cell wall. 3) ClpL
inhibits pneumococcal adherence to A549 cells through activation
of the small Rho-GTPases via Rap1 up-regulation.
ClpL-induced Rap1 activates Rac1, which in turn phosphorylates cofilin and inactivates actin
cytoskeleton rearrangement. These findings may shed light on how the HSP100 family
member ClpL can modulate virulence at the early stages of infection. Taken
together, genes induced by pneumococcal invasion or by the host cells could be
valuable biomarkers for diagnosis, preventive, and therapeutic measures to
overcome pneumococcal diseases.
Dong-Kwon Rhee, Ph.D. (Email: dkrhee@skku.edu)
Professor (Molecular Microbiology Lab, School of Pharmacy)
Research
Interests
Pneumococcal
pathogenesis, Host-cell invasion, Signaling, Host-pathogen interaction, Stress
response
7 (OU-4)
A novel antimicrobial agent
from Nuphar japonicum
Teruo KURODA, Associate Professor
Department
of Environmental and Applied Microbiology
[Introduction]
The
emergence of drug resistant bacteria is a growing clinical problem that can
cause therapeutic failures throughout the world. In particular, methicillin-resistant
Staphylococcus aureus (MRSA) has recently
become one of the most important pathogenic bacteria. To develop anti-MRSA
drugs, we have been screening active compounds which have anti-MRSA activity,
and investigate their action mechanisms.
[Methods]
The active compound named “compound K” was isolated
from methanol extract of Nuphar rhizome (the rhizome of Nuphar japonicum) with liquid-liquid separation and silica gel
column chromatography. Minimum inhibitory concentration of compound K was
determined microdilution method according to the recommendations of the
Japanese Society of Chemotherapy.
[Results & Discussion]
Compound K showed potent growth inhibition of S. aureus. MICs of compound K against
several MRSA and methicillin-sensitive S.
aureus (MSSA) were 1-4 mg/mL. In addition, compound K showed antimicrobial
activity for vancomycin intermediate-resistant S. aureus (VISA). It indicated that compound K was effective
against S. aureus irrespective of
sensitivity to oxacillin and vancomycin. When compound K was added at
one-fourth concentration of MIC, MICs of gentamicin and/or arbekacin were
significantly decreased. This synergy effect was observed for several clinical
isolated MRSA. Compound K also showed inhibition for topoisomerase IV of S. aureus which have essential role in
DNA replication. However, compound K did not exhibit cross-resistance to
norfloxacin-resistant S. aureus, which indicated that the action site in topoisomerase IV was different
from that of quinolone resistance-determining regions (QRDR) and that it
inhibited other targets besides topoisomerase IV.
Teruo KURODA, Ph.D. (Email:
tkuroda@cc.okayama-u.ac.jp)
Associate Professor (Department
of Environmental and Applied Microbiology)
Research
Interests
MRSA,
antimicrobial agents, antiseptic agents, multidrug efflux pump, Pseudomonas aeruginosa
8 (SKKU-4)
Organic-inorganic
hybrid nanoparticles for siRNA delivery
Ji Hoon Jeong, Associate Professor
School
of Pharmacy, Sungkyunkwan University, Republic of Korea
Gene therapy based on small interference RNA
(siRNA) holds enormous potential for therapeutic intervention of a broad range
of genetic diseases, including infectious diseases, gene-related disorders, and
cancer. A number of cationic carriers forming nanoparticulates mostly via
electrostatic interactions with oppositely charged nucleic acid have been
developed for siRNA delivery. However, due to its rigid double-stranded
structure, siRNA has often failed to generate nanoparticulates as tight as
those formed with plasmid DNA with conventional cationic carriers. Therefore,
improving transfection efficiency siRNA is prerequisite for its successful use
in clinical settings. Inorganic materials such as hydroxyapatite and gold have
a wide range of applications in biomedical systems. In this study, we
demonstrated bio-organic templated inorganic nanoparticles for siRNA delivery.
The new modalities of the organic-inorganic hybrid nanoparticles would give a
chance to cope with the siRNA encapsulation problem and enable concurrent
cellular siRNA delivery and imaging. The possibilities of using the
nanoparticles for therapeutic and imaging purposes were assessed.
Ji Hoon Jeong, Ph.D. (Email: jhjeong@skku.edu)
Associate Professor (School of Pharmacy)
Research
Interests
gene
and drug delivery, drug dissolution and stabilization, microneedles, stem cell
engineering
9
(OU-5)
Mast
Cell: A critical regulator of inflammation
Satoshi
Tanaka
Professor, Immunobiology
[Introduction]
Mast
cells are found in nearly all the vascularized tissues, although the
physiological roles of mast cells remain to be fully clarified. Accumulating
evidence indicates that mast cells play critical roles in inflammatory
responses, such as immediate allergy, whereas mast cells have recently been
found to be involved in suppression of immune responses, such as immune
tolerance. Mast cells originate in the hematopoietic stem cells in the bone
marrow and undergo terminal differentiation in the tissues, in which they are
ultimately resident. The characteristics of tissue mast cells are, therefore,
profoundly affected by their microenvironmental factors, including the
neighboring cells, extracellular matrix, and local cytokine/growth factor
levels. Although recent studies demonstrated using the genetically-mast
cell-deficient models that mast cells should be required for a variety of
physiological and pathological responses, it remains unknown how tissue mast
cells regulate these responses.
[Results and Discussion]
We
established a culture model of murine cutaneous mast cells, in which
IL-3-dependent bone marrow-derived cultured mast cells (BMMCs) are co-cultured
with Swiss 3T3 fibroblasts in the presence of stem cell factor (Takano et al., FEBS Lett., 2008). This model reflects
the characteristics of cutaneous mast cells, such as sensitivity to substance P
and increased granule storage of histamine, heparin and neutral proteases. We
extracted genes, of which expression levels were drastically changed during the
co-culture period. We focused on the up-regulation of CD44, one of the primary
receptor for hyaluronan, because hyaluronan is abundant in the cutaneous
tissues and might be associated with cutaneous mast cells. We demonstrated that
CD44 regulates the process of proliferation of mast cells in the cutaneous
tissues (Takano et al., Lab. Invest., 2009). We previously found the impaired granule maturation in the mature
peritoneal mast cells obtained from the gene-targeted mice that lacks the
ability of histamine synthesis. By using our cutaneous mast cell model,
we revealed that histamine synthesis should be required for granule maturation
of mast cells, of which process is largely independent of specific histamine
receptors (Nakazawa et al., Eur. J.
Immunol., 2013). Our findings will contribute to development of novel therapeutic
compounds that prevent proliferation and maturation of tissue mast cells
for chronic inflammatory diseases.
Satoshi
Tanaka, Ph.D. (Email:
tanaka@ pharm.okayama-u.ac.jp)
Professor, Department of Immunobiology
Research
Interests
Mast
Cell, Inflammation, Allergy, Histamine, Microenvironment