| 1. |
Fadlina Aulia, Hiroaki Matsuba, Shoya Adachi, Takumi Yamada, Ikuhiko Nakase, Teruki Nii, Takeshi Mori, Yoshiki Katayama, and Akihiro Kishimura, Effective design of PEGylated polyion complex (PIC) nanoparticles for enhancing PIC internalisation in cells utilising block copolymer combinations with mismatched ionic chain lengths., J. Mater. Chem B., 10.1039/D3TB02049E, 12, 1826-1836, 2024.02, In nanomedicine, PEGylation of nanomaterials poses a dilemma since it inhibits their interaction with target cells and enables their retention in target tissues despite its biocompatibility and nonspecific internalisation suppression. PEGylated polypeptide-based polyion complexes (PICs) are fabricated via the self-assembly of PEGylated aniomers and homocatiomers based on electrostatic interactions. We propose that various parameters like block copolymer design and PIC domain characteristics can enhance the cell–PEGylated PIC interactions. Remarkably, the properties of the PIC domain were tuned by the matched/mismatched ionomer chain lengths, PIC domain crosslinking degree, chemical modification of cationic species after crosslinking, PIC morphologies (vesicles/micelles) and polyethylene glycol (PEG) chain lengths. Cellular internalisation of the prepared PICs was evaluated using HeLa cells. Consequently, mismatched ionomer chain lengths and vesicle morphology enhanced cell–PIC interactions, and the states of ion pairing, particularly cationic residues, affected the internalisation behaviours of PICs via acetylation or guanidinylation of amino groups on catiomers. This treatment attenuated the cell–PIC interactions, possibly because of reduced interaction of PICs with negatively charged species on the cell-surface, glycosaminoglycans. Moreover, morphology and PEG length were correlated with PIC internalisation, in which PICs with longer and denser PEG were internalised less effectively. Cell line dependency was tested using RAW 264.7 macrophage cells; PIC recognition could be maintained after capping amino groups on catiomers, indicating that the remaining anionic groups were still effectively recognised by the scavenger receptors of macrophages. Our strategy for tuning the physicochemical properties of the PEGylated PIC nanocarriers is promising for overcoming the PEG issue.. |
| 2. |
Chuya Tateishi, Akihiro Koga, Atsuhiro Matsuura, Ryosuke Kaneko, Kenta Tanito, Teruki Nii, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, A human cell orthogonal enzyme β-D-galacturonidase for sensitive detection of antigen proteins, Analyst, 10.1039/d3an00314k, 2023.04. |
| 3. |
Riku Saeki, Shingo Kobayashi, Rena Shimazui, Teruki Nii, Akihiro Kishimura, Takeshi Mori, Masaru Tanaka, Yoshiki Katayama, Characterization of polypropyleneimine as an alternative transfection reagent, Analytical Sciences, 10.1007/s44211-023-00284-x, 2023.03. |
| 4. |
Kenta Tanito, Teruki Nii, Yuta Yokoyama, Haruka Oishi, Mayuka Shibata, Shoichi Hijii, Ryosuke Kaneko, Chuya Tateishi, Shoko Ito, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Engineered macrophages acting as a trigger to induce inflammation only in tumor tissues, Journal of Controlled Release, 10.1016/j.jconrel.2023.04.010, 2023.04. |
| 5. |
Biplab K C, Teruki Nii, Takeshi Mori, Yoshiki Katayama, Akihiro Kishimura, Dynamic frustrated charge hotspots created by charge density modulation sequester globular proteins into complex coacervates, Chemical Science, 10.1039/d3sc00993a, 14, 24, 6608-6620, 2023.05, This study presents a simple strategy for the sequestration of globular proteins as clients into synthetic polypeptide-based complex coacervates as a scaffold, thereby recapitulating the scaffold-client interaction found in biological condensates. Considering the low net charges of scaffold proteins participating in biological condensates, the linear charge density (s) on the polyanion, polyethylene glycol-b-poly(aspartic acids), was reduced by introducing hydroxypropyl or butyl moieties as a charge- neutral pendant group. Complex coacervate prepared from the series of reduced-s polyanions and the polycation, homo-poly-L-lysine, could act as a scaffold that sequestered various globular proteins with high encapsulation efficiency (>80%), which sometimes involved further agglomerations in the coacervates. The sequestration of proteins was basically driven by electrostatic interaction, and therefore depended on the ionic strength and charges of the proteins. However, based on the results of polymer partitioning in the coacervate in the presence or absence of proteins, charge ratios between cationic and anionic polymers were maintained at the charge ratio of unity. Therefore, the origin of the electrostatic interaction with proteins is considered to be dynamic frustrated charges in the complex coacervates created by non-neutralized charges on polymer chains. Furthermore, fluorescence recovery after photobleaching (FRAP) measurements showed that the interaction of side-chains and proteins changed the dynamic property of coacervates. It also suggested that the physical properties of the condensate are tunable before and after the sequestration of globular proteins. The present rational design approach of the scaffold-client interaction is helpful for basic life-science research and the applied frontier of artificial organelles.. |
| 6. |
Akinori Goto, Yasutaka Anraku, Shigeto Fukushima and Akihiro Kishimura, Increased Enzyme Loading in PICsomes Via Controlling Membrane Permeability Improves Enzyme Prodrug Cancer Therapy Outcome, Polymers, https://doi.org/10.3390/polym15061368, 15, 6, 1368, 2023.03. |
| 7. |
Asmariah Ahmad, Tomoki Maruyama, Teruki Nii, Takeshi Mori, Yoshiki Katayama, and Akihiro Kishimura, Facile preparation of hexagonal nanosheets via polyion complex formation from α-helical polypeptides and polyphosphate-based molecules, Chemical Communications, 10.1039/d2cc05137k, 59, 12, 1657-1660, 2023.01. |
| 8. |
Takashi Miyano, Koichiro Kato, Ryosuke Kaneko, Toshihide Kimura, Misa Maruoka, Akihiro Kishimura, Michiko Furuta, and Yoshihisa Yamashita, Dietary Problems Are Associated with Frailty Status in Older People with Fewer Teeth in Japan, International Journal of Environmental Research and Public Health, https://doi.org/10.3390/ijerph192316260, 19, 16260, 2022.12. |
| 9. |
Asmariah Ahmad, Teruki Nii, Takeshi Mori, Yoshiki Katayama, Masanori Toyofuku, Akihiro Kishimura, Nanostructure Control of an Antibiotic-Based Polyion Complex Using a Series of Polycations with Different Side-Chain Modification Rates., Macromolecular rapid communications, 10.1002/marc.202200316, e2200316, 2022.06, Developing nanovehicles for delivering antibiotics is a promising approach to overcome the issue of antibiotic resistance. This study aims to utilize a polyion complex (PICs) system for developing novel nanovehicles for polymyxin-type antibiotics, which are known as last resort drugs. The formation of antibiotic-based PIC nanostructures is investigated using colistimethate sodium (CMS), an anionic cyclic short peptide, and a series of block catiomers bearing different amounts of guanidinium moieties on their side chains. In addition, only the modified catiomer, and not the unmodified catiomer, self-assembles with CMS, implying the importance of the guanidine moieties for enhancing the interaction between the catiomer and CMS via the formation of multivalent hydrogen bonding. Moreover, micellar and vesicular PIC nanostructures are selectively formed depending on the ratio of the guanidine residues. Size-exclusion chromatography reveals that the encapsulation efficiency of CMS is dependent on the guanidinium modification ratio. The antimicrobial activity of the PIC nanostructures is also confirmed, indicating that the complexation of CMS in the PICs and further release from the PICs successfully occurs.. |
| 10. |
Haitao Feng, Jeong-Hun Kang, Song Qi, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Preparation of a PEGylated liposome that co-encapsulates l-arginine and doxorubicin to achieve a synergistic anticancer effect., RSC advances, 10.1039/d1ra06514a, 11, 54, 34101-34106, 2021.10, Strategies that combine chemotherapies with unconventional agents such as nitric oxide (NO) have been shown to enhance cancer therapies. Compared with small molecule chemotherapy drugs, nanosized particles have improved therapeutic efficacies and reduced systemic side effects because of the enhanced permeability and retention effect. In this report, we prepared PEGylated liposomes (LP) that incorporated l-arginine (Arg) and the anticancer drug doxorubicin (Dox) to yield a co-delivery system (Dox-Arg-LP). On the basis of our previous research, we hypothesized that Dox-Arg-LP should achieve a synergistic anticancer effect because Arg conversion to NO by activated M1 macrophages augments the chemotherapeutic activity of Dox. Dox-Arg-LP showed comparable physical properties to those of conventional Dox-only liposomes (Dox-LP). In vitro assessment revealed that the cytotoxicity of Dox-Arg-LP toward cancer cells was significantly higher than that of Dox-LP. In vivo application of Dox-Arg-LP in mice enhanced the chemotherapeutic effect with a 2 mg kg-1 dose of Dox-Arg-LP achieving the same therapeutic efficacy as a two-fold higher dose of Dox-LP (i.e., 4 mg kg-1). Therefore, co-encapsulation of dual agents into a liposome formulation is an efficient strategy to enhance chemotherapy while reducing systemic toxicity.. |
| 11. |
Shunyi Li, Kyosuke Yakabe, Khadijah Zai, Yiwei Liu, Akihiro Kishimura, Koji Hase, Yun-Gi Kim, Takeshi Mori, Yoshiki Katayama, Specific adsorption of a β-lactam antibiotic in vivo by an anion-exchange resin for protection of the intestinal microbiota., Biomaterials science, 10.1039/d1bm00958c, 9, 21, 7219-7227, 2021.10, The fraction of antibiotics that are excreted from the intestine during administration leads to disruption of commensal bacteria as well as resulting in dysbiosis and various diseases. To protect the gut microbiota during treatment with antibiotics, use of activated carbon (AC) has recently been reported as a method to adsorb antibiotics. However, the antibiotic adsorption by AC is nonspecific and may also result in the adsorption of essential biological molecules. In this work, we reported that an anion exchange resin (AER) has better specificity than AC for adsorbing the β-lactam antibiotic cefoperazone (CEF). Because CEF has a negatively charged carboxylate group and a conjugated system, the AER was used to adsorb CEF through electrostatic and π-π interactions. The AER was specific for CEF over biological molecules such as bile acids and vitamins in the intestine. The AER protected Escherichia coli from CEF in vitro. Furthermore, oral administration of the AER reduced the fecal free CEF concentration, and protected the gut microbiota from CEF-induced dysbiosis.. |
| 12. |
Ryosuke Kaneko, Tsuyoshi Oda, Ryosuke Yoshida, Chuya Tateishi, Kenta Tanito, Teruki Nii, Akihiro Kishimura, Noriho Kamiya, Takeshi Mori, Yoshiki Katayama, alpha-L-Arabinofuranosidase as an Orthogonal Enzyme for Human Cells, CHEMISTRY LETTERS, 10.1246/cl.210231, 50, 8, 1493-1495, 2021.08, Here we proposed a-L-arabinofuranosidase from Clostridium thermocellum as an enzyme for signal amplification in bioanalysis whose activity does not exist in human cells. Combination of this enzyme and beta-galactosidase enabled simultaneous detection of two different antigens on live cells.. |
| 13. |
Chan Woo Kim, Riki Toita, Jeong-Hun Kang, Takeshi Mori, Akihiro Kishimura, Yoshiki Katayama, Protein Kinase C alpha-Responsive Gene Carrier for Cancer-Specific Transgene Expression and Cancer Therapy, ACS BIOMATERIALS SCIENCE & ENGINEERING, 10.1021/acsbiomaterials.1c00213, 7, 6, 2530-2537, 2021.06, The presence of intracellular signal transduction and its abnormal activities in many cancers has potential for medical and pharmaceutical applications. We recently developed a protein kinase C alpha (PKC alpha)-responsive gene carrier for cancerspecific gene delivery. Here, we demonstrate an in-depth analysis of cellular signal-responsive gene carrier and the impact of its selective transgene expression in response to malfunctioning intracellular signaling in cancer cells. We prepared a novel gene carrier consisting of a linear polyethylenimine (LPEI) main chain grafted to a cationic PKC alpha-specific substrate (FKKQGSFAKKK-NH2). The LPEI-peptide conjugate formed a nanosized polyplex with pDNA and mediated efficient cellular uptake and endosomal escape. This polyplex also led to successful transgene expression which responded to the target PKC alpha in various cancer cells and exhibited a 10-100-fold higher efficiency compared to the control group. In xenograft tumor models, the LPEI-peptide conjugate promoted transgene expression showing a clear-cut response to PKC alpha. Furthermore, when a plasmid containing a therapeutic gene, human caspase-8 (pcDNA-hcasp8), was used, the LPEIpeptide conjugate had significant cancer-suppressive effects and extended animal survival. Collectively, these results reveal that our method has great potential for cancer-specific gene delivery and therapy.. |
| 14. |
Kenta Tanito, Yunosuke Oshiro, Hiroshi Tagawa, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Comparative Evaluation of Natural Killer Cell-Mediated Cell Killing Assay Based on the Leakage of an Endogenous Enzyme or a Pre-Loaded Fluorophore., Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 10.2116/analsci.21P117, 2021.05. |
| 15. |
Jinting Li, Yunmei Mu, Yiwei Liu, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Effect of Size and Loading of Retinoic Acid in Polyvinyl Butyrate Nanoparticles on Amelioration of Colitis, POLYMERS, 10.3390/polym13091472, 13, 9, 2021.05, Butyrate has been used in the treatment of inflammatory bowel diseases (IBD). However, the controlled release of butyrate has been indicated to be necessary in order to avoid the side effects verified at high concentrations. We previously developed nanoparticles (NPs) of polyvinyl butyrate (PVBu) as an oral butyrate donor for the controlled release of butyrate for the treatment of colitis. To examine the effect of the size of NPs on the therapeutic effect of colitis, here we prepared PVBu NPs with different sizes (100 nm and 200 nm). Both sizes of PVBu NPs significantly suppressed the inflammatory response in macrophages in vitro. PVBu NPs with 200 nm showed better effects on the amelioration of colitis compared with the 100 nm-NPs. We found unexpectedly that 200 nm-NP incorporated with all-trans retinoic acid (ATRA) showed a much better therapeutic effect than those with unloaded 200 nm-NPs, although ATRA alone was reported to worsen the inflammation. The synergistic effect of ATRA with butyrate shows evidence of being a promising approach for IBD treatment.. |
| 16. |
Yiwei Liu, Tomoki Maruyama, K. C. Biplab, Takeshi Mori, Yoshiki Katayama, Akihiro Kishimura, Inducible Dynamic Behavior of Polyion Complex Vesicles by Disrupting Charge Balance, CHEMISTRY LETTERS, 10.1246/cl.210037, 50, 5, 1034-1037, 2021.05, The formation and dynamic behavior of polyion complex-based vesicles (PICsomes) were examined under charge-balance disrupting conditions, using homo-catiomer and pegylated-aniomer. Under catiomer-rich conditions, an unusual increase in the PICsome size was observed, which was primarily attributed to Ostwald ripening. This growth was triggered by the addition of a homo-catiomer to switch the composition of the PIC membrane. The results provide insight into the design of smart PIC-based systems for functional nanocapsules.. |
| 17. |
Kaneko, Ryosuke; Kawamura, Masumi; Kishimura, Akihiro; Mori, Takeshi; Katayama, Yoshiki, Effect of a Chloroacetyl Modification on the Suppression of Dissociation of a Fluorescent Molecule from Cells for Antigen- Specific Cell Staining, ANALYTICAL SCIENCES, 10.2116/analsci.20SCN03, 37, 3, 529-532, 2021.03. |
| 18. |
Takanobu Nobori, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, A FRET-based Protein Kinase Assay using Phos-tag-modified Quantum Dots and Fluorophore-labeled Peptides., Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 10.2116/analsci.20P443, 2021.03. |
| 19. |
Ryosuke Kaneko, Masumi Kawamura, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Effect of a Chloroacetyl Modification on the Suppression of Dissociation of a Fluorescent Molecule from Cells for Antigen- Specific Cell Staining, ANALYTICAL SCIENCES, 10.2116/analsci.20SCN03, 37, 3, 529-532, 2021.03, We previously developed a hydrolase-based fluorescence amplification method for antigen-specific cell labelling, in which fluorescent substrates stained cells by a non-covalent hydrophobic interaction. To improve the substrates retention in cells, we examined the effect of a chloroacetyl group modification on the substrate retention. We found that the chloroacetyl group suppressed the dissociation of the substrate after forming a covalent bond with intracellular proteins. However, the slow reaction speed of the chloroacetyl group allowed dissociation for cells in the early stage of the staining reaction.. |
| 20. |
Yunmei Mu, Yusuke Kinashi, Jinting Li, Takuma Yoshikawa, Akihiro Kishimura, Mitsuru Tanaka, Toshiro Matsui, Takeshi Mori, Koji Hase, Yoshiki Katayama, Polyvinyl Butyrate Nanoparticles as Butyrate Donors for Colitis Treatment, ACS APPLIED BIO MATERIALS, 10.1021/acsabm.0c01105, 4, 3, 2335-2341, 2021.03, Butyrate has been attracting attention for the suppression of inflammatory bowel disease (IBD). However, clinical trials of butyrate for IBD treatment have resulted in controversial outcomes, likely owing to the adverse effect of butyrate on the intestinal epithelium that was observed at high butyrate concentrations. Herein, we propose polyvinyl butyrate (PVBu) nanoparticles (NPs) as butyrate donors for delivery to the lower part of the intestine for the treatment of colitis. The PVBu NPs suppressed the inflammatory activation of macrophages in vitro, although sodium butyrate inversely further activated macrophages. Oral administration of NPs did not change the luminal concentration of free butyrate; however, NPs showed a therapeutic effect on a colitis mouse model. In addition, incorporation of vitamin D-3 into the NPs enhanced the therapeutic effect on colitis. Hence, PVBu NPs are a promising therapeutic for IBD treatment, not only as a butyrate donor but also as a carrier for hydrophobic drugs like vitamin D-3.. |
| 21. |
Koichi Sasaki, Kyohei Muguruma, Rento Osawa, Akane Fukuda, Atsuhiko Taniguchi, Akihiro Kishimura, Yoshio Hayashi, Takeshi Mori, Yoshiki Katayama, Synthesis and biological evaluation of a monocyclic Fc-binding antibody-recruiting molecule for cancer immunotherapy, RSC MEDICINAL CHEMISTRY, 10.1039/d0md00337a, 12, 3, 406-409, 2021.03, Antibody-recruiting molecules (ARMs) are bispecific molecules composed of an antibody-binding motif and a target-binding motif that redirect endogenous antibodies to target cells to elicit immune responses. To enhance the translational potential of ARMs, it is crucial to design antibody/target-binding motifs that have strong affinity and are easy to synthesize. Here, we synthesized a novel Fc-binding ARM (Fc-ARM) that targets folate receptor (FR)-positive cancer cells, Reo-3, using a recently developed monocyclic peptide 15-Lys8Leu, which binds strongly to the Fc region of an antibody. Reo-3 bound to the Fc region of the antibody with a K-d of 5.8 nM, and recruited a clinically used antibody mixture to attack FR-positive IGROV-1 cells as efficiently as Fc-ARM2, in which a bicyclic Fc-binding peptide was used. These results indicate that 15-Lys8Leu, which can be synthesized readily, is suitable for various applications including the development of Fc-ARMs.. |
| 22. |
Koichi Sasaki, Minori Harada, Takuma Yoshikawa, Hiroshi Tagawa, Yui Harada, Yoshikazu Yonemitsu, Takaaki Ryujin, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Fc-Binding Antibody-Recruiting Molecules Targeting Prostate-Specific Membrane Antigen: Defucosylation of Antibody for Efficacy Improvement**, CHEMBIOCHEM, 10.1002/cbic.202000577, 22, 3, 496-500, 2021.02, Synthetic small molecules that redirect endogenous antibodies to target cells are promising drug candidates because they overcome the potential shortcomings of therapeutic antibodies, such as immunogenicity and the need for intravenous delivery. Previously, we reported a novel class of bispecific molecules targeting the antibody Fc region and folate receptor, named Fc-binding antibody-recruiting molecules (Fc-ARMs). Fc-ARMs can theoretically recruit most endogenous antibodies, inducing antibody-dependent cell-mediated cytotoxicity (ADCC) to eliminate cancer cells. Herein, we describe new Fc-ARMs that target prostate cancer (Fc-ARM-Ps). Fc-ARM-Ps recruited antibodies to cancer cells expressing prostate-specific membrane antigen but did so with lower efficiency compared with Fc-ARMs targeting the folate receptor. Upon recruitment by Fc-ARM-P, defucosylated antibodies efficiently activated natural killer cells and induced ADCC, whereas antibodies with intact N-glycans did not. The results suggest that the affinity between recruited antibodies and CD16a, a type of Fc receptor expressed on immune cells, could be a key factor controlling immune activation in the Fc-ARM strategy.. |
| 23. |
Xizheng Sun, Reika Tokunaga, Yoko Nagai, Ryo Miyahara, Akihiro Kishimura, Shigeru Kawakami, Yoshiki Katayama, Takeshi Mori, Ligand Design for Specific MHC Class I Molecules on the Cell Surface, BIOCHEMISTRY, 10.1021/acs.biochem.0c00735, 59, 49, 4646-4653, 2020.12, We have validated that ligand peptides designed from antigen peptides could be used for targeting specific major histocompatibility complex class I (MHC-I) molecules on the cell surface. To design the ligand peptides, we used reported antigen peptides for each MHC-I molecule with high binding affinity. From the crystal structure of the peptide/MHC-I complexes, we determined a modifiable residue in the antigen peptides and replaced this residue with a lysine with an e-amine group modified with functional molecules. The designed ligand peptides successfully bound to cells expressing the corresponding MHC-I molecules via exchange of peptides bound to MHC-I. We demonstrated that the peptide ligands could be used to transport a protein or a liposome to cells expressing the corresponding MHC-I. This strategy may be useful for targeted delivery to cells overexpressing MHC-I, which have been observed in autoimmune diseases.. |
| 24. |
Yunmei Mu, Yusuke Kinashi, Akihiro Kishimura, Takeshi Mori, Koji Hase, Yoshiki Katayama, Effect of polyvinyl butyrate nanoparticles incorporated with immune suppressing vitamins on alteration of population of intestinal immune cells, PROGRESS IN NATURAL SCIENCE-MATERIALS INTERNATIONAL, 10.1016/j.pnsc.2020.10.003, 30, 5, 707-710, 2020.10, The nanoparticles (NPs) from polyvinyl butyrate (PVBu) which can work as an orally applicable donor of butyrate for intestine were prepared. Immunotolerance inducing molecules such as vitamin D-3 and all-trans retinoic acid (ATRA) were incorporated into PVBu NPs. The alteration in populations and numbers of DC103(+) dendritic cells (DC) and regulatory T (Treg) cells in intestinal immune tissues were examined after oral administration of NPs. It was found that NPs reduced the population of CD103(+) DC and Treg cells in Peyer's patched in lower part of the intestine and inversely increased the population of CD103(+) DC in mesenteric lymph node (MLN), while the population of Treg cells in MLN was unchanged. These observations indicate that NPs may enhance the immunotolerance at MLN and lamina propria toward luminal antigens, indicating the promising features of PVBu NPs as therapeutics of allergy and autoimmune diseases.. |
| 25. |
Haitao Feng, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Evaluation of a Synergistic Effect of L-Arginine on the Anticancer Activity of Doxorubicin by Using a Co-culture System, ANALYTICAL SCIENCES, 10.2116/analsci.20P200, 36, 10, 1279-1283, 2020.10, In the early stage of tumor development, tumor-associated macrophages (TAM) works to suppress tumor growth by secreting soluble factors including nitric oxide (NO). L-Arginine (Arg) is a substrate of nitric oxide synthase (NOS) expressed in TAM. Here we examined whether NO produced from Arg by macrophages works to enhance the effect of the anti-cancer drug, doxorubicin (Dox) by using a co-culture system of cancer cells with macrophages. By employing colorimetric analyses methods (Griess Reagent and Cell Counting kit-8), we found that NO produced from Arg by co-cultured macrophages could enhance the cytotoxic effect of Dox to cancer cells. Moreover, we found that augmentation is affected by the order of the addition of Arg and Dox. A prior addition of Arg to Dox and simultaneous addition showed the same enhancement effect, but a prior addition of Dox to Arg abolished the augmentation. This suggests that the co-administration of Arg with Dox would be an effective treatment to improve chemo-therapies.. |
| 26. |
Kazuki Yuzuriha, Ayaka Yoshida, Shunyi Li, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Synthesis of peptide conjugates with vitamins for induction of antigen-specific immunotolerance, JOURNAL OF PEPTIDE SCIENCE, 10.1002/psc.3275, 26, 10, 2020.10, In this report, we designed conjugates of an antigen peptide with the immunosuppressive vitamins all-transretinoic acid (ATRA) and vitamin D3 for efficient induction of antigen-specific immunotolerance. We established a synthetic scheme for the preparation of the peptide-vitamin conjugates, which the chemically unstable vitamins tolerated. Among the obtained conjugates, the ATRA conjugate successfully suppressed inflammatory effects in macrophages and dendritic cells and induced antigen presentation in dendritic cells. This synthetic method of conjugate is conceivably applicable to other antigen peptides for induction of antigen-specific immunotolerance.. |
| 27. |
Beob Soo Kim, Mitsuru Naito, Hiroyuki Chaya, Mao Hori, Kotaro Hayashi, Hyun Su Min, Yu Yi, Hyun Jin Kim, Tetsuya Nagata, Yasutaka Anraku, Akihiro Kishimura, Kazunori Kataoka, and Kanjiro Miyata, Noncovalent Stabilization of Vesicular Polyion Complexes with Chemically Modified/Single-Stranded Oligonucleotides and PEG-b-guanidinylated Polypeptides for Intracavity Encapsulation of Effector Enzymes Aimed at Cooperative Gene Knockdown, Biomacromolecules, 10.1021/acs.biomac.0c01192, 21, 10, 4365-4376, 2020.09, For the simultaneous delivery of antisense oligonucleotides and their effector enzymes into cells, nanosized vesicular polyion complexes (PICs) were fabricated from oppositely charged polyion pairs of oligonucleotides and poly(ethylene glycol) (PEG)-b-polypeptides. First, the polyion component structures were carefully designed to facilitate a multimolecular (or secondary) association of unit PICs for noncovalent (or chemical cross-linking-free) stabilization of vesicular PICs. Chemically modified, single-stranded oligonucleotides (SSOs) dramatically stabilized the multimolecular associates under physiological conditions, compared to control SSOs without chemical modifications and duplex oligonucleotides. In addition, a high degree of guanidino groups in the polypeptide segment was also crucial for the high stability of multimolecular associates. Dynamic light scattering and transmission electron microscopy revealed the stabilized multimolecular associates to have a 100 nm sized vesicular architecture with a narrow size distribution. The loading number of SSOs per nanovesicle was determined to be ∼2500 using fluorescence correlation spectroscopic analyses with fluorescently labeled SSOs. Furthermore, the nanovesicle stably encapsulated ribonuclease H (RNase H) as an effector enzyme at ∼10 per nanovesicle through simple vortex-mixing with preformed nanovesicles. Ultimately, the RNase H-encapsulated nanovesicle efficiently delivered SSOs with RNase H into cultured cancer cells, thereby eliciting the significantly higher gene knockdown compared with empty nanovesicles (without RNase H) or a mixture of nanovesicles with RNase H without encapsulation. These results demonstrate the great potential of noncovalently stabilized nanovesicles for the codelivery of two varying bio-macromolecule payloads for ensuring their cooperative biological activity.. |
| 28. |
Yunmei Mu, Jinting Li, Jeong-Hun Kang, Hinako Eto, Khadijah Zai, Akihiro Kishimura, Fuminori Hyodo, Takeshi Mori, Yoshiki Katayama, A Lipid-Based Nanocarrier Containing Active Vitamin D-3 Ameliorates NASH in Mice via Direct and Intestine-Mediated Effects on Liver Inflammation, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 10.1248/bob.b20-00432, 43, 9, 1413-1420, 2020.09, The gut liver axis may be involved in non-alcoholic steatohepatitis (NASH) progression. Pathogen-associated molecular patterns leak through the intestinal barrier to the liver via the portal vein to contribute to NASH development. Active vitamin D-3 (1,25(OH)(2)D-3) is a potential therapeutic agent to enhance the intestinal barrier. Active vitamin D-3 also suppresses inflammation and fibrosis in the liver. However, the adverse effects of active vitamin D-3 such as hypercalcemia limit its clinical use. We created a nano-structured lipid carrier (NLC) containing active vitamin D-3 to deliver active vitamin D-3 to the intestine and liver to elicit NASH treatment. We found a suppressive effect of the NLC on the lipopolysaccharide-induced increase in permeability of an epithelial layer in vitro. Using mice in which NASH was induced by a methionine and choline-deficient diet, we discovered that oral application of the NLC ameliorated the permeability increase in the intestinal barrier and attenuated steatosis, inflammation and fibrosis in liver at a safe dose of active vitamin D-3 at which the free form of active vitamin D-3 did not show a therapeutic effect. These data suggest that the NLC is a novel therapeutic agent for NASH.. |
| 29. |
Haitao Feng, Le Thanh Nam, Takuma Yoshikawa, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Effect of an Endothelin B Receptor Agonist on the Tumor Accumulation of Nanocarriers, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 10.1248/bpb.b20-00367, 43, 9, 1301-1305, 2020.09, Enhancing blood flow to tumors is a prominent strategy for improving the tumor accumulation of macromolecular drugs through the enhanced permeability and retention (EPR) effect. IRL-1620 is an agonist of the endothelin B receptor, and is a promising molecule to enhance tumor blood flow by activating endothelial nitric oxide synthase. However, contradictory effects on tumor blood flow modulation have been reported because the effects of IRL-1620 may differ in different animal models. Here, we examined for the first time the effect of IRL-1620 on the EPR effect for PEGylated liposomes in a CT-26 murine colon cancer model. Co-injection of IRL-1620 at an optimum dose (3nmol/kg) nearly doubled the tumor accumulation of liposomes compared with controls, indicating that IRL-1620 enhanced the EPR effect in the present colon cancer model. Co-injection of IRL-1620 is a promising strategy to improve the therapeutic effects of macromolecular drugs while reducing their side effects.. |
| 30. |
Beob Soo Kim, Mitsuru Naito, Rimpei Kamegawa, Hyun Jin Kim, Ryo Iizuka, Takashi Funatsu, Shingo Ueno, Takanori Ichiki, Akihiro Kishimura and Kanjiro Miyata, Photo-reactive oligodeoxynucleotide-embedded nanovesicles (PROsomes) with switchable stability for efficient cellular uptake and gene knockdown, CHEMICAL COMMUNICATIONS, 10.1039/d0cc01750g, 56, 66, 9477-9480, 2020.08. |
| 31. |
Ryujin, T, Shimizu, T, Miyahara, R, Asai, D, Shimazui, R, Yoshikawa, T, Kishimura, A, Mori, T, Ishida, T, Katayama, Y, Blood retention and antigenicity of polycarboxybetaine-modified liposomes, Int. J. Pharm., in press, 2020.06. |
| 32. |
Takuma Yoshikawa, Khanh Quoc Phan, Hiroshi Tagawa, Koichi Sasaki, Haitao Feng, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Modification of nitric oxide donors onto a monoclonal antibody boosts accumulation in solid tumors, INTERNATIONAL JOURNAL OF PHARMACEUTICS, 10.1016/j.ijpharm.2020.119352, 583, 119352-119352, 2020.06, Although monoclonal antibodies (mAbs) have revolutionized cancer treatment, their accumulation in solid tumors is limited and requires improvement to enhance therapeutic efficacy. Here we developed a strategy to modify mAb with a donor of nitric oxide (NO) because NO functions to vasodilate as well as to enhance the permeability of vascular endothelium, which will contribute to enhancing the tumor accumulation of mAb. We selected S-nitrosothiol as a NO donor and established the procedure to modify S-nitrosothiol group on mAb under ambient conditions. The modified mAb (Ab-SNO) thus obtained released NO in a preferable speed and maintained its original properties such as binding affinity to a target antigen and efficacy to induce antibody-dependent cellular cytotoxicity. We demonstrated that Ab-SNO enhanced the tumor accumulation of co-administered proteins such as antibody and serum albumin.. |
| 33. |
Hiroshi Tagawa, Katsuya Maruyama, Koichi Sasaki, Natsuki Konoue, Akihiro Kishimura, Motomu Kanai, Takeshi Mori, Kounosuke Oisaki, Yoshiki Katayama, Induction of ADCC by a folic acid–mAb conjugate prepared by tryptophan-selective reaction toward folate-receptor-positive cancer cells, RSC Advances, 10.1039/d0ra03291c, 10, 28, 16727-16731, 2020.04, 有機ラジカルを用いたトリプトファン選択的反応によりモノクローナル後退（mAb）-葉酸複合体を常温下で調製したところ、抗体依存性細胞傷害性（ADCC）の有意な誘導能を示した。. |
| 34. |
Koichi Sasaki, Minori Harada, Yoshiki Miyashita, Hiroshi Tagawa, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Fc-binding antibody-recruiting molecules exploit endogenous antibodies for anti-tumor immune responses, Chemical Science, 10.1039/d0sc00017e, 11, 12, 3208-3214, 2020.03, Redirecting endogenous antibodies in the bloodstream to tumor cells using synthetic molecules is a promising approach to trigger anti-tumor immune responses. However, current molecular designs only enable the use of a small fraction of endogenous antibodies, limiting the therapeutic potential. Here, we report Fc-binding antibody-recruiting molecules (Fc-ARMs) as the first example addressing this issue. Fc-ARMs are composed of an Fc-binding peptide and a targeting ligand, enabling the exploitation of endogenous antibodies through constant affinity to the Fc region of antibodies, whose sequence is conserved in contrast to the Fab region. We show that Fc-ARM targeting folate receptor-α (FR-α) redirects a clinically used antibody mixture to FR-α+ cancer cells, resulting in cancer cell lysis by natural killer cells in vitro. Fc-ARMs successfully interacted with antibodies in vivo and accumulated in tumors. Furthermore, Fc-ARMs recruited antibodies to suppress tumor growth in a mouse model. Thus, Fc-ARMs have the potential to be a novel class of cancer immunotherapeutic agents.. |
| 35. |
穆 云妹, 李 晋廷, Zai Khadijah, 岸村 顕広, 兵藤 文紀, 森 健, 片山 佳樹, ビタミンD3含有脂質ナノ粒子によるリーキーガットの改善に基づくNASHの治療(Therapeutic effect of lipid-based nanocarriers containing vitamin D3 on NASH by ameliorating the leaky gut), 日本薬学会年会要旨集, 140年会, 27X-pm12S, 2020.03. |
| 36. |
Takanobu Nobori, Masumi Kawamura, Ryosuke Yoshida, Taisei Joichi, Kenta Kamino, Akihiro Kishimura, Eishi Baba, Takeshi Mori, Yoshiki Katayama, Fluorescence Signal Amplification by Using β-Galactosidase for Flow Cytometry; Advantages of an Endogenous Activity-Free Enzyme, Analytical chemistry, 10.1021/acs.analchem.9b04471, 92, 4, 3069-3076, 2020.02, We previously proposed using a hydrolysis enzyme for fluorescent signal amplification in flow cytometric detection of antigen proteins, which was named the catalyzed reporter penetration (CARP) method. In this method, antigen proteins are labeled with enzyme-modified antibodies, and then fluorophore-modified substrates stain cells by penetrating the cell membrane upon hydrolysis of the substrate. We proved the concept by using alkaline phosphatase (AP) as the hydrolysis enzyme. However, a required prior inactivation process of endogenous AP activity on the cell surface risked disrupting recognition of antigen proteins by antibodies. In this report, the CARP method was extended to β-galactosidase (β-gal) as an amplification enzyme, which circumvented the requirement of an initial inactivation process because endogenous β-gal activity on the surface of examined cells was found to be negligible. The substrate structure for β-gal was optimized and used for the CARP method. The CARP method showed significantly higher fluorescent signals than a conventional method using fluorophore-modified antibodies. Moreover, the degree of amplification of the fluorescence signal was higher for antigens with low expression levels, showing that the CARP method is a suitable signal amplification method over current conventional approaches.. |
| 37. |
Takanobu Nobori, Masumi Kawamura, Ryosuke Yoshida, Taisei Joichi, Kenta Kamino, Akihiro Kishimura, Eishi Baba, Takeshi Mori, Yoshiki Katayama, Fluorescence Signal Amplification by Using β-Galactosidase for Flow Cytometry; Advantages of an Endogenous Activity-Free Enzyme., Analytical chemistry, 10.1021/acs.analchem.9b04471, 92, 4, 3069-3076, 2020.02, We previously proposed using a hydrolysis enzyme for fluorescent signal amplification in flow cytometric detection of antigen proteins, which was named the catalyzed reporter penetration (CARP) method. In this method, antigen proteins are labeled with enzyme-modified antibodies, and then fluorophore-modified substrates stain cells by penetrating the cell membrane upon hydrolysis of the substrate. We proved the concept by using alkaline phosphatase (AP) as the hydrolysis enzyme. However, a required prior inactivation process of endogenous AP activity on the cell surface risked disrupting recognition of antigen proteins by antibodies. In this report, the CARP method was extended to β-galactosidase (β-gal) as an amplification enzyme, which circumvented the requirement of an initial inactivation process because endogenous β-gal activity on the surface of examined cells was found to be negligible. The substrate structure for β-gal was optimized and used for the CARP method. The CARP method showed significantly higher fluorescent signals than a conventional method using fluorophore-modified antibodies. Moreover, the degree of amplification of the fluorescence signal was higher for antigens with low expression levels, showing that the CARP method is a suitable signal amplification method over current conventional approaches.. |
| 38. |
Hiroko Nagai, Wataru Hatanaka, Masayoshi Matsuda, Akihiro Kishimura, Yoshiki Katayama, Takeshi Mori, Folate receptor-specific cell-cell adhesion by using a folate-modified peptide-based anchor., Journal of biomaterials science. Polymer edition, 10.1080/09205063.2019.1616975, 30, 11, 983-993, 2019.08, We report here a folate-modified membrane anchor for cell surface modification to induce cell adhesion to target cells. The membrane anchor region, which was consisted of cationic lysine residues and palmitoyl group-modified residues, was modified with folate through an oligoethlene glycol linker. The peptide anchor was modified on to the cell membrane by using β-cyclodextrin as a solubilizer of the peptide anchor. After modification, the peptide anchor disappeared from the cell membrane via endocytotic uptake or dissociation from the cell membrane. However, the endocytosed peptide was represented on the cell surface via recycling endosome pathway. The obtained folate-modified cells successfully adhered on to target cells which expressed folate receptor α via ligand-receptor specific interaction and adhesion continued at least 4 hours.. |
| 39. |
Hiroko Nagai, Wataru Hatanaka, Masayoshi Matsuda, Akihiro Kishimura, Yoshiki Katayama, Takeshi Mori, Folate receptor-specific cell-cell adhesion by using a folate-modified peptide-based anchor, Journal of Biomaterials Science, Polymer Edition, 10.1080/09205063.2019.1616975, 30, 11, 983-993, 2019.07, We report here a folate-modified membrane anchor for cell surface modification to induce cell adhesion to target cells. The membrane anchor region, which was consisted of cationic lysine residues and palmitoyl group-modified residues, was modified with folate through an oligoethlene glycol linker. The peptide anchor was modified on to the cell membrane by using β-cyclodextrin as a solubilizer of the peptide anchor. After modification, the peptide anchor disappeared from the cell membrane via endocytotic uptake or dissociation from the cell membrane. However, the endocytosed peptide was represented on the cell surface via recycling endosome pathway. The obtained folate-modified cells successfully adhered on to target cells which expressed folate receptor α via ligand-receptor specific interaction and adhesion continued at least 4 hours.. |
| 40. |
Takuma Yoshikawa, Yukina Mori, Haitao Feng, Khanh Quoc Phan, Akihiro Kishimura, Jeong-Hun Kang, Takeshi Mori, Yoshiki Katayama, Rapid and continuous accumulation of nitric oxide-releasing liposomes in tumors to augment the enhanced permeability and retention (EPR) effect, INTERNATIONAL JOURNAL OF PHARMACEUTICS, 10.1016/j.ijpharm.2019.05.043, 565, 481-487, 2019.06, The modulation of blood flow to tumors is a prominent strategy for improving the tumor accumulation of nanomedicines, resulting from the enhanced permeability and retention (EPR) effect. We previously reported a promising EPR enhancer-a nitric oxide (NO) donor-containing liposome (NO-LP)-which showed enhanced accumulation in tumor tissue. Herein, we study NO-LP in greater detail to clarify its practical use as an EPR enhancer. NO-LP was found to have advantages as a NO donor, including the ability to maintain NO donation over long periods of time, and a constant rate of NO-release irrespective of the environmental pH. NO-LP showed rapid accumulation in tumor tissue after injection (1 h), and then accumulation was continuously enhanced until 48 h. Enhanced NO-LP accumulation was observed specifically in tumor, while the accumulation in other organs remained relatively unchanged. The results obtained show the promising features of NO-LP as an EPR enhancer.. |
| 41. |
Yiwei Liu, Takeshi Mori, Yoshiki Katayama, Akihiro Kishimura, Yolk-shell assembly formation based on polyion complex of proteins, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 257, 2019.03. |
| 42. |
Kim BS, Chuanoi S, Suma T, Anraku Y, Hayashi K, Naito M, Kim HJ, Kwon IC, Miyata K, Kishimura A, Kataoka K, Self-Assembly of siRNA/PEG- b-Catiomer at Integer Molar Ratio into 100 nm-Sized Vesicular Polyion Complexes (siRNAsomes) for RNAi and Codelivery of Cargo Macromolecules., Journal of the American Chemical Society, 10.1021/jacs.8b13641, 141, 8, 3699-3709, 2019.02. |
| 43. |
Khadijah Zai, Narumi Ishihara, Hiroyuki Oguchi, Masato Hirota, Akihiro Kishimura, Takeshi Mori, Koji Hase, Yoshiki Katayama, Regulation of inflammatory response of macrophages and induction of regulatory T cells by using retinoic acid-loaded nanostructured lipid carrier, JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION, 10.1080/09205063.2018.1493671, 30, 1, 1-11, 2019.01, Immunomodulatory function of all-trans retinoic acid (ATRA) has been gathering much attention for the therapy of autoimmune diseases. ATRA is a chemically unstable molecule which requires proper formulation for targeted delivery. Here we examined nanostructured lipid carrier (NLC) for the formulation of ATRA. NLC is a representative nanoparticle formulation especially suited for oral delivery. We established the preparation procedures of ATRA-containing NLC (NLC-RA) which minimizes the degradation of ATRA during the preparation process. NLC-RA thus obtained was taken up by macrophages and induced anti-inflammatory response via suppressing NF-kappa B signaling as well as via enhancing the production of anti-inflammatory cytokines. Moreover, NLC-RA enhanced differentiation of naive T cells to regulatory T cells in the co-culture system with dendritic cells. These results suggest that NLC-RA is a promising alternative therapy for the autoimmune diseases especially intestinal bowel disease.[GRAPHICS].. |
| 44. |
Wataru Hatanaka, Hiroki Takeuchi, Minaho Koga, Taka Aki Ryujin, Akihiro Kishimura, Yoshiki Katayama, Shinya Tsukiji, Takeshi Mori, Synthesis of transmembrane molecules by click chemistry, Chemistry Letters, 10.1246/cl.190009, 48, 5, 433-436, 2019.01, We report here efficient synthesis of artificial transmembrane molecules (ATMs), via click chemistry of a transmembrane part and membrane-penetrating parts. Synthesized ATMs penetrated the cell membrane by Coulombic force, while they stayed on the cell membrane once they were complexed with streptavidin.. |
| 45. |
Masayuki Yokoyama, Kouichi Shiraishi, Akihiro Kishimura, Glassware cleaning, Drug Delivery System, 10.2745/dds.34.213, 34, 3, 213-215, 2019.01. |
| 46. |
Khadijah Zai, Kazuki Yuzuriha, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Preparation of Complexes between Ovalbumin Nanoparticles and Retinoic Acid for Efficient Induction of Tolerogenic Dendritic Cells, ANALYTICAL SCIENCES, 10.2116/analsci.18P252, 34, 11, 1243-1248, 2018.11, The induction of antigen-specific immunotolerance has been gathering attention concerning the therapy of allergy and autoimmune diseases. Tolerogenic dendritic cells (tDCs) play crucial roles in immunotolerance therapy because they induce anergic responses for auto-reactive helper T cells, and also enhance differentiation to regulatory T cells to maintain tolerance against auto-antigens. All-trans retinoic acid (ATRA) is one of the representative molecules used to induce tDCs. We have proposed a simple formulation of ovalbumin nanoparticles complexed with ATRA (OVA/RA NPs). OVA/RA NPs were taken up by DCs and successfully induced phenotypes of tDCs.. |
| 47. |
Hori Mao, Cabral Horacio, Toh Kazuko, Kishimura Akihiro, Kataoka Kazunori, Robust Polyion Complex Vesicles (PICsomes) under Physiological Conditions Reinforced by Multiple Hydrogen Bond Formation Derived by Guanidinium Groups, BIOMACROMOLECULES, 10.1021/acs.biomac.8b01097, 19, 10, 4113-4121, 2018.10. |
| 48. |
Masamitsu Suhara, Yutaka Miura, Horacio Cabral, Daisuke Akagi, Yasutaka Anraku, Akihiro Kishimura, Masaya Sano, Takuya Miyazaki, Noriko Nakamura, Ayako Nishiyama, Kazunori Kataoka, Hiroyuki Koyama, Katsuyuki Hoshina, Targeting ability of self-assembled nanomedicines in rat acute limb ischemia model is affected by size, JOURNAL OF CONTROLLED RELEASE, 10.1016/j.jconrel.2018.07.049, 286, 394-401, 2018.09, Peripheral artery disease (PAD) is one of the most spreading diseases all over the world. The treatment strategies are limited to surgical or endovascular procedures for final stage chronic PAD or acute limb ischemia, and no pharmacological approaches have been achieved to prevent the worsening of chronic PAD or to regenerate the tissues of acute limb ischemia. Therefore, the improvement of therapeutic strategy is strongly demanded in clinics. Here, we adopted an acute hindlimb ischemia model in rats, which provides concomitant inflammatory response, to evaluate the application of drug delivery system against PAD. Through comparative experiments by using different-sized nanomedicine analogues, polyion complex (PIC) micelles with 30 nm diameter and PIC vesicles with 100- and 200-nm diameter (PICs-30, -100, -200 respectively), we found the size-dependent accumulation and retention in the collateral arteries. In contrast to PICs-30 and -200, histological analysis showed that PICs-100 were around the arterioles and co-localized with macrophages, which indicates that the PICs-100 can achieve moderate interaction with phagocytes. Our data suggests that controlling the size of nanomedicines has promise for developing novel angiogenic treatments toward the effective management of collateral arteries.. |
| 49. |
Khadijah Zai, Masato Hirota, Takahiro Yamada, Narumi Ishihara, Takeshi Mori, Akihiro Kishimura, Koichiro Suzuki, Koji Hase, Yoshiki Katayama, Therapeutic effect of vitamin D-3-containing nanostructured lipid carriers on inflammatory bowel disease, JOURNAL OF CONTROLLED RELEASE, 10.1016/j.jconrel.2018.07.019, 286, 94-102, 2018.09, The active form of vitamin D-3, 1,25(OH)(2)D-3 has been found to exert multiple effects on the suppression of progression of inflammatory bowel disease (IBD). Vitamin D-3 has been gathering attention as a therapy for IBD. However, the clinical trials conducted to date revealed that a relatively high dosage of vitamin D-3 was required to see a significant therapeutic effect. Thus, effective formulation and delivery of vitamin D-3 to colonic inflammatory lesions will be required. Herein we describe the preparation of a nanostructured lipid carrier (NLC) for the encapsulation of 1,25(OH)(2)D-3 for colonic delivery via oral administration. The optimized fabrication procedure enabled the incorporation of 1,25(OH)(2)D-3 in the NLC by minimizing the destruction of chemically unstable 1,25(OH)(2)D-3. The obtained NLCs orally delivered 1,25(OH)(2)D-3 to the colon in mice and maintained a high concentration of 1,25(OH)(2)D-3 in the colonic tissue for at least 12 h. The NLC showed multiple effects on the suppression of symptoms of colitis induced by dextran sodium sulfate, namely maintaining crypt structure, reducing the tissue concentration of inflammatory cytokines, suppressing the infiltration of polymorphonuclear leukocytes, and augmenting anti-inflammatory CX3CR1high macrophages. Our NLCs containing 1,25(OH)(2)D-3 may be an alternative treatment for IBD therapy.. |
| 50. |
Hikari Sato, Yoshiki Miyashita, Koichi Sasaki, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Non-covalent Coating of Liposome Surface with IgG through Its Constant Region, CHEMISTRY LETTERS, 10.1246/cl.180181, 47, 6, 770-772, 2018.06, Here we present a liposome non-covalently coated with immunoglobulin G (IgG) by using a peptide ligand to the constant region of IgG. IgG coated on the liposome still maintained binding ability to a complementary antigen protein but it was not recognized by Fc gamma receptors (Fc gamma Rs) of monocytes. The stealth feature of the IgG-coated liposome may be useful to avoid clearance by macrophages in vivo to extend the blood half-life of the liposome.. |
| 51. |
Koichi Sasaki, Yoshiki Miyashita, Daisuke Asai, Daiki Funamoto, Kazuki Sato, Yoko Yamaguchi, Yuji Mishima, Tadafumi Iino, Shigeo Takaishi, Jun Nagano, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, A peptide inhibitor of antibody-dependent cell-mediated cytotoxicity against EGFR/folate receptor-α double positive cells., MedChemComm, 10.1039/c8md00010g, 9, 5, 783-788, 2018.05, Antibody-dependent cell-mediated cytotoxicity (ADCC) is caused by natural killer (NK) cells upon recognition of antigen-bound IgG via FcγRIIIa. This mechanism is crucial for cytolysis of pathogen-infected cells and monoclonal antibody (mAb)-mediated elimination of cancer cells. However, there is concern that mAb-based cancer therapy induces ADCC against non-target cells expressing antigens. To date, no strategy has been reported to enhance the selectivity of ADCC to protect non-target cells expressing antigens. Here, we introduce a model inhibitor which specifically blocks ADCC of anti-EGFR mAbs towards EGFR/folate receptor α (FRα) double positive cells. This inhibitor recruits mAbs on the FRα of the cell surface independent of Fab antigen recognition. The resulting ternary and/or quaternary complexes formed on the cell surface suppress signal transduction of FcγRIIIa in NK cells, consequently leading to more specific ADCC.. |
| 52. |
効率的な光線力学療法を指向したハイブリッド型光増感剤送達システム. |
| 53. |
Hikari Sato, Elnaz Nakhaei, Takahito Kawano, Masaharu Murata, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Ligand-Mediated Coating of Liposomes with Human Serum Albumin., Langmuir : the ACS journal of surfaces and colloids, 10.1021/acs.langmuir.7b04024, 34, 6, 2324-2331, 2018.02, Coating liposome surfaces with human serum albumin (HSA) can improve the colloidal stability and prevent opsonization. HSA coating via specific binding with alkyl ligands is promising because although the ligand-mediated coating is relatively stable it can spontaneously exchange with fresh HSA. However, to achieve surface coating with HSA, multiple hydrophobic ligands must be exposed to an aqueous medium prior to binding with HSA. This presents a challenge, as hydrophobic ligands tend to be buried in the liposomal membrane. Here we present the first HSA modification of liposome surfaces via alkyl ligands. We found that a relatively short alkyl ligand, or a long alkyl ligand with a terminal carboxylate, could be exposed on the liposome surface without causing aggregation of the liposomes and these ligands could subsequently bind HSA. The resulting HSA-coated liposomes were as inert as conventional PEGylated liposomes in terms of macrophage recognition.. |
| 54. |
Martin J. Hollamby, Catherine F. Smith, Melanie M. Britton, Ashleigh E. Danks, Zoe Schnepp, Isabelle Grillo, Brian R. Pauw, Akihiro Kishimura, Takashi Nakanishi, The aggregation of an alkyl-C60 derivative as a function of concentration, temperature and solvent type, Physical Chemistry Chemical Physics, 10.1039/c7cp06348b, 20, 5, 3373-3380, 2018.02, Contrast-variation small-angle neutron scattering (CV-SANS), small-angle X-ray scattering (SAXS), nuclear magnetic resonance (NMR) measurements of diffusion and isothermal titration calorimetry (ITC) are used to gain insight into the aggregation of an alkyl-C60 derivative, molecule 1, in n-hexane, n-decane and toluene as a function of concentration and temperature. Results point to an associative mechanism of aggregation similar to other commonly associating molecules, including non-ionic surfactants or asphaltenes in non-aqueous solvents. Little aggregation is detected in toluene, but small micelle-like structures form in n-alkane solvents, which have a C60-rich core and alkyl-rich shell. The greatest aggregation extent is found in n-hexane, and at 0.1 M the micelles of 1 comprise around 6 molecules at 25 °C. These micelles become smaller when the concentration is lowered, or if the solvent is changed to n-decane. The solution structure is also affected by temperature, with a slightly larger aggregation extent at 10 °C than at 25 °C. At higher concentrations, for example in solutions of 1 above 0.3 M in n-decane, a bicontinuous network becomes apparent. Overall, these findings aid our understanding of the factors driving the assembly of alkyl-π-conjugated hydrophobic amphiphiles such as 1 in solution and thereby represent a step towards the ultimate goal of exploiting this phenomenon to form materials with well-defined order.. |
| 55. |
Elnaz Nakhaei, Ko Takehara, Hikari Sato, Khadijah Zai, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, A dual alkylated peptide-ligand enhances affinity to human serum albumin, analytical sciences, 10.2116/analsci.17P614, 34, 4, 501-504, 2018.01, Therapeutic peptides and diagnostic agents with their molecular size below the renal clearance threshold suffer from short blood circulation time. Here, we report a novel design of peptide-based ligand with a strong binding affinity to human serum albumin (HSA), which can be used as a tag to extend the blood circulation of small-size molecules. We designed ligands with dual alkyl groups connected with a negatively charged spacer. The ligands showed both higher binding affinity to HSA and a higher retention in mice blood than that of a single alkylated peptide.. |
| 56. |
Takanobu Nobori, Kenta Tosaka, Akira Kawamura, Taisei Joichi, Kenta Kamino, Akihiro Kishimura, Eishi Baba, Takeshi Mori, Yoshiki Katayama, Alkaline Phosphatase-Catalyzed Amplification of a Fluorescence Signal for Flow Cytometry., Analytical chemistry, 10.1021/acs.analchem.7b03893, 90, 2, 1059-1062, 2018.01, Despite the expanding use of flow cytometry, its detection limit is not satisfactory for many antigen proteins with low copy numbers. Herein, we describe an alkaline phosphatase (AP)-based technique to amplify the fluorescence signal for cell staining applications. We designed a fluorescent substrate that acquires membrane permeability upon dephosphorylation by AP. By using the substrate, the fluorescence signal of cells in flow cytometry could be successfully amplified to give a much stronger signal than the cells labeled using a conventional fluorophore-modified antibody.. |
| 57. |
Omer F. Mutaf, Yasutaka Anraku, Akihiro Kishimura, Kazunori Kataoka, Unilamellar polyion complex vesicles (PICsomes) with tunable permeabilities for macromolecular solutes with different shapes and sizes, POLYMER, 10.1016/j.polymer.2017.10.062, 133, 1-7, 2017.12, Polyion complex vesicles (PICsomes) are characterized by their unique three-layered semipermeable nanomembrane structures, in which a unilamellar PIC layer is sandwiched by poly(ethylene glycol) layers, and have gathered much attention as nano-scaled drug vehicles. Herein, the crosslinking degree of the nanomembrane in the PICsome was controlled systematically for the first time. Permeability of the PICsome nanomembrane was evaluated through a kinetic study of the release of macromolecular cargoes from the PICsome. The degree of crosslinking in the nanomembrane successfully regulated the release behavior. Moreover, the shape and size of the macromolecular solutes were found to be critical factors determining their transport from the inner aqueous phase of the PICsome to the external environment. The results indicate that the unique three-layered structure of PICsome membranes plays a key role in modulating solute transport. These findings will provide a rational strategy for the development of nanomembrane-based controlled-release systems. (C) 2017 Elsevier Ltd. All rights reserved.. |
| 58. |
Yuta Nakamura, Hikari Sato, Takanobu Nobori, Hotaru Matsumoto, Shoko Toyama, Tomohiro Shuno, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Modification of ligands for serum albumin on polyethyleneimine to stabilize polyplexes in gene delivery, Journal of Biomaterials Science, Polymer Edition, 10.1080/09205063.2017.1328730, 28, 13, 1382-1393, 2017.09, In this work, we have developed a new technique to stabilize a ternary complex composed of plasmid DNA, a linear polyethyleneimine (LPEI) and serum albumin. A stearoyl group was conjugated to LPEI as a specific ligand for serum albumin. The resultant ternary complex has excellent stability in physiological saline conditions, maintaining its initial diameter and preventing aggregation of red blood cells. The ternary complex has equivalent transfection ability to and significantly lower cytotoxicity than unmodified LPEI. Therefore, the ternary complex is potentially useful as a new gene carrier, possessing high blood stability.. |
| 59. |
Hengmin Tang, Yuki Sakamura, Takeshi Mori, Yoshiki Katayama, Akihiro Kishimura, Development of Enzyme Loaded Polyion Complex Vesicle (PICsome): Thermal Stability of Enzyme in PICsome Compartment and Effect of Coencapsulation of Dextran on Enzyme Activity, MACROMOLECULAR BIOSCIENCE, 10.1002/mabi.201600542, 17, 8, 2017.08, Applications of enzymes are intensively studied, particularly for biomedical applications. However, encapsulation or immobilization of enzymes without deactivation and long-term use of enzymes are still at issue. This study focuses on the polymeric vesicles "PICsomes" for encapsulation of enzymes to develop a hecto-nanometer-scaled enzyme-loaded reactor. The catalytic activity of a PICsome-based enzyme nanoreactor is carefully examined to clarify the effect of compartmentalization by PICsome. Encapsulation by PICsome provides a stability enhancement of enzymes after 24 h incubation at 37 degrees C, which is particularly helpful for maintaining the high effective concentration of beta-galactosidase. Moreover, to control the microenvironment inside the nanoreactor, a large amount of dextran, a neutral macromolecule, is encapsulated together with beta-galactosidase in the PICsome. The resulting dextran-coloaded nanoreactor contributes to the enhancement of enzyme stability, even after exposure to 24 h incubation at -20 degrees C, mainly due to the antifreezing effect.. |
| 60. |
Enzyme-catalyzed amplification of fluorescent immunolabeling of a single cell for high-sensitive flow cytometry. |
| 61. |
Md. Zahangir Hosain, Kazuki Yuzuriha, Khadijah, Masafumi Takeo, Akihiro Kishimura, Yoshihiko Murakami, Takeshi Mori, Yoshiki Katayama, Synergic modulation of the inflammatory state of macrophages utilizing anti-oxidant and phosphatidylserine-containing polymer-lipid hybrid nanoparticles, MEDCHEMCOMM, 10.1039/c7md00174f, 8, 7, 1514-1520, 2017.07, Inflammatory activation of macrophages is a key factor in chronic inflammatory diseases such as ulcerative colitis. The excessive production of reactive oxygen species (ROS)/reactive nitrogen species (RNS) by macrophages causes oxidative stress during the inflammatory response and exaggerates inflammatory lesions in ulcerative colitis. Inhibition of the inflammatory activation of macrophages is a promising treatment for chronic inflammatory diseases. Here, we prepared self-filling polymer-lipid hybrid nanoparticles (PST-PLNPs) consisting of poly DL-lactic acid as a hydrophobic biodegradable polymer core encapsulating alpha-tocopherol (T) and phosphatidylserine (PS) both on the surface and interior of the particle. We confirmed the anti-inflammatory response of these hybrid nanoparticles in activated murine macrophages. PS has anti-inflammatory effects on macrophages by modulating the macrophage phenotype, while alpha-tocopherol is an antioxidant that neutralizes ROS. We found that PS-containing (PS-PLNPs) and PS plus alpha-tocopherol-containing (PST-PLNPs) polymer-lipid hybrid nanoparticles significantly increased the viability of lipopolysaccharide (LPS)-treated macrophages compared with phosphatidylcholine-containing PLNPs. PST-PLNPs had a better effect than PS-PLNPs, which was attributed to the synergy between PS and alpha-tocopherol. This synergic action of PST-PLNPs reduced NO and pro-inflammatory cytokine (IL-6) production and increased anti-inflammatory cytokine (TGF-beta 1) production when incubated with activated macrophages. Thus, these self-filling biodegradable polymer-lipid hybrid nanoparticles (PST-PLNPs) containing antioxidant and anti-inflammatory molecules might be potential alternative drug carriers to liposomes and polymeric nanoparticles for the treatment of chronic inflammatory diseases such as ulcerative colitis.. |
| 62. |
Elnaz Nakhaei, Chan Woo Kim, Daiki Funamoto, Hikari Sato, Yuta Nakamura, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Design of a ligand for cancer imaging with long blood circulation and an enhanced accumulation ability in tumors, MEDCHEMCOMM, 10.1039/c7md00102a, 8, 6, 1190-1195, 2017.06, The use of imaging agents to visualize tumor cells is an advantageous technique to achieve a more efficient intraoperative diagnosis and effective debulking operations. Targeting of these agents to certain receptors that are overexpressed in cancer cells, such as the folate receptor, aids in tumor targeting. Several imaging probes have been developed using this strategy. However, these ligand-targeting cancer imaging probes are rapidly cleared during systemic delivery due to their small size, which compromises their biodistribution and circulation. Improving the detection of cancer requires higher accumulation and effective retention activities of imaging probes. Here we developed a new design for a folate-fluorophore conjugate that is modified with palmitoyl. Palmitic acid has a strong binding affinity with human serum albumin (HSA), which has the ability to form non-covalent host-guest complexes and has a blood half-life of 19 days. In this strategy, HSA is expected to serve as an endogenous nanocarrier for the designed probe in blood circulation. We hypothesized that via a reversible interaction with HSA, this simple palmitoyl modification on a folate-fluorophore conjugate can induce long blood circulation of the probe. Our folate-targeted probe could show longer blood circulation compared to the probe which lacks palmitoyl.. |
| 63. |
Cuicui Li, Masafumi Takeo, Masayoshi Matsuda, Hiroko Nagai, Sun Xizheng, Wataru Hatanaka, Akihiro Kishimura, Hiroyuki Inoue, Kenzaburo Tani, Takeshi Mori, Yoshiki Katayama, Facilitating the presentation of antigen peptides on dendritic cells for cancer immunotherapy using a polymer-based synthetic receptor, MEDCHEMCOMM, 10.1039/c7md00188f, 8, 6, 1207-1212, 2017.06, The introduction of proteins into dendritic cells (DCs) ex vivo is a critical step for the DC-based immunotherapy of cancer. Here, we developed a biotin-modified polymer with multiple hydrophobic membrane anchors for cells that functions as a synthetic receptor for an antigen protein, ovalbumin (OVA), to introduce it efficiently into DCs compared with the conventional pulsing method. Our method showed significant advantages, including the rapid incorporation of OVA and the activation of antigen-specific T cells in a MHC-restricted manner. When mice were immunized by DCs treated with our method, tumor growth was completely suppressed, indicating that our method can be used to prepare adjuvant DCs.. |
| 64. |
Akinori Goto, Hui Chi Yen, Yasutaka Anraku, Shigeto Fukushima, Ping-Shan Lai, Masaru Kato, Akihiro Kishimura, Kazunori Kataoka, Facile Preparation of Delivery Platform of Water-Soluble Low-Molecular-Weight Drugs Based on Polyion Complex Vesicle (PlCsome) Encapsulating Mesoporous Silica Nanoparticle, ACS BIOMATERIALS SCIENCE & ENGINEERING, 10.1021/acsbiomaterials.6b00562, 3, 5, 807-815, 2017.05, Polyion complex vesicles (PlCsomes) are polymeric hollow capsules composed of a unique semipermeable membrane, which may represent a versatile platform for constructing drug-loaded nanoformulation. However, it is difficult to retain water-soluble low-molecular weight compounds (LMWCs) in the inner space of PlCsome because of the high permeability of PIC membrane for LMWCs. Herein, we selected mesoporous silica nanoparticle (MSN) as a drug-retaining nanomatrix, and we demonstrated successful encapsulation of MSN into the PlCsome to obtain MSN@PlCsome. The efficacy of MSN loading, a ratio of the amount of MSN encapsulated in the PICsome to the amount of feed MSN, was at most 83%, and the diameter of resulting product was approximately 100 nm. The obtained MSN@PlCsome was stably dispersed under the physiological condition, and showed considerable longevity in blood circulation of mice. Furthermore, the surface of MSN in MSN@PlCsome can be modified without any deterioration of the vesicle structure, obtaining amino-functionalized and sulfonate-functionalized MSN@PlCsomes (A-MSN@PlCsome and S-MSN@PICsome, respectively). Both surface-modified MSN@PlCsomes were successfully loaded with charged water-soluble low-molecular-weight compounds (LMWCs). Particularly, S-MSN@PICsome kept 8 wt % gemcitabine (GEM) per S-MSN, and released it in a sustained manner. GEM-loaded S-MSN@PlCsome demonstrated marked cytotoxicity against cultured tumor cells, and achieved significant in vivo efficacy to suppress the growth of subcutaneously implanted lung tumor via intravenous administration.. |
| 65. |
Wataru Hatanaka, Miki Kawaguchi, Xizheng Sun, Yusuke Nagao, Hiroyuki Ohshima, Mitsuru Hashida, Yuriko Higuchi, Akihiro Kishimura, Yoshiki Katayama, Takeshi Mori, Use of Membrane Potential to Achieve Transmembrane Modification with an Artificial Receptor, BIOCONJUGATE CHEMISTRY, 10.1021/acs.bioconjchem.6b00449, 28, 2, 296-301, 2017.02, We developed a strategy to modify cell membranes with an artificial transmembrane receptor. Coulomb force on the receptor, caused by the membrane potential, was used to achieve membrane penetration. A hydrophobically modified cationic peptide was used as a membrane potential sensitive region that was connected to biotin through a transmembrane oligoethylene glycol (OEG) chain. This artificial receptor gradually disappeared from the cell membrane via penetration despite the presence of a hydrophilic OEG chain. However, when the receptor was bound to streptavidin (SA), it remained on the cell membrane because of the large and hydrophilic nature of SA.. |
| 66. |
Yu Tahara, Takuma Yoshikawa, Hikari Sato, Yukina Mori, Md Hosain Zahangir, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Encapsulation of a nitric oxide donor into a liposome to boost the enhanced permeation and retention (EPR) effect, MEDCHEMCOMM, 10.1039/c6md00614k, 8, 2, 415-421, 2017.02, We propose a method to improve the enhanced permeability and retention (EPR) effect of nanomedicines based on tumor-specific vasodilation using a nitric oxide (NO) donor-containing liposome. NONOate, a typical NO donor, was incorporated into a PEGylated liposome to retard the protonation-induced release of NO from NONOate by the protecting lipid bilayer membrane. The NONOate-containing liposome (NONOate-LP) showed similar blood retention to an empty PEGylated liposome but almost twice the amount accumulated within the tumor. This improvement in the EPR effect is thought to have been caused by specific vasodilation in the tumor tissue by NO released from the NONOate-LP accumulated in the tumor. The improved EPR effect by NONOate-LP will be useful for the accumulation of co-administered nanomedicines.. |
| 67. |
Naoki Sasaki, Mariko Tatanou, Tomoko Suzuki, Yasutaka Anraku, Akihiro Kishimura, Kazunori Kataoka, Kae Sato, A Membrane-integrated Microfluidic Device to Study Permeation of Nanoparticles through Straight Micropores toward Rational Design of Nanomedicines, ANALYTICAL SCIENCES, 10.2116/analsci.32.1307, 32, 12, 1307-1314, 2016.12, Nanoparticles have been widely utilized to deliver drugs from blood vessels to target tissues. A crucial issue concerning nanoparticle-based drug delivery is to discuss the relationship between experimentally-obtained permeability and physical parameters. Although nanoparticles can permeate vascular pores, because the size and shape of the pores are essentially non-uniform, conventional animal testing and recent cell-based microfluidic devices are unable to precisely evaluate the effects of physical parameters (e.g. pore size and nanoparticle size) on permeation. In this study, we present a membrane integrated microfluidic device to study permeation of nanoparticles through straight micropores. Porous membranes possessing uniform straight pores were utilized. The effects of pore size and pressure difference across the pores on nanoparticle permeation were examined. The experimentally determined permeability coefficient of 1.0 mu m-pore membrane against 100 nm-diameter nanoparticles agreed well with the theoretical value obtained for convectional permeation. Our method can be utilized to clarify the relationship between the experimentally-obtained permeability and physical parameters, and will help rational design of nanomedicines.. |
| 68. |
次世代医療を支えるバイオマテリアルのイノベーション 酸応答性無機有機ハイブリッド型光増感剤デリバリーシステム. |
| 69. |
Kenshiro Naoyama, Takeshi Mori, Yoshiki Katayama, Akihiro Kishimura, Fabrication of Dendrimer-Based Polyion Complex Submicrometer-Scaled Structures with Enhanced Stability under Physiological Conditions, MACROMOLECULAR RAPID COMMUNICATIONS, 10.1002/marc.201600171, 37, 13, 1087-1093, 2016.07, Submicrometer-scaled (sub-) self-assembled materials have been developed based on polyion complex (PIC) formation, in particular for biomedical-applications. However, sufficient stability under physiological conditions is required for their practical use. In this study, PIC formation behavior is examined using a block aniomer, poly(ethylene glycol)-b-poly(aspartic acid), and homocatiomers, poly(l-lysine) (LPK) and dendritic poly(l-lysine) (DPK) with different generations, to elucidate the contribution of the dendritic architecture to stability enhancement. LPK-based PIC shows a sub-vesicular structure only at 25 degrees C in the absence of NaCl; in contrast, DPK-based PIC forms a sub-structure under physiological salt concentration and temperature conditions, even when the number of charges of a single molecule is much smaller than that of LPK. Moreover, the formation of sub-vesicular and -spherical micellar structures is dependent on DPK generation. Thus, the molecular backbone architecture of the PIC component plays an important role not only in expanding the preparation conditions and enhancing stability, but also in controlling the self-assembled structures, mainly due to the spatially restricted structures of dendrimers.. |
| 70. |
Ryosuke Taniguchi, Yutaka Miura, Hiroyuki Koyama, Tsukasa Chida, Yasutaka Anraku, Akihiro Kishimura, Kunihiro Shigematsu, Kazunori Kataoka, Toshiaki Watanabe, Adequately-Sized Nanocarriers Allow Sustained Targeted Drug Delivery to Neointimal Lesions in Rat Arteries, MOLECULAR PHARMACEUTICS, 10.1021/acs.molpharmaceut.6b00219, 13, 6, 2108-2116, 2016.06, In atherosclerotic lesions, the endothelial barrier against the bloodstream can become compromised, resulting in the exposure of the extracellular matrix (ECM) and intimal cells beneath. In theory, this allows adequately sized nanocarriers in circulation to infiltrate into the intimal lesion intravascularly. We sought to evaluate this possibility using rat carotid arteries with induced neointima. Cy5-labeled polyethylene glycol-conjugated polyion complex (PIC) micelles and vesicles, with diameters of 40, 100, or 200 nm (PICs-40, PICs-100, and PICs-200, respectively) were intravenously administered to rats after injury to the carotid artery using a balloon catheter. High accumulation and long retention of PICs-40 in the induced neointima was confirmed by in vivo imaging, while the accumulation of PICs-100 and PICs-200 was limited, indicating that the size of nanocarriers is a crucial factor for efficient delivery. Furthermore, epirubicin-incorporated polymeric micelles with a diameter similar to that of PICs-40 showed significant curative effects in rats with induced neointima, in terms of lesion size and cell number. Specific and effective drug delivery to pre-existing neointimal lesions was demonstrated with adequate size control of the nanocarriers. We consider that this nanocarrier-based drug delivery system could be utilized for the treatment of atherosclerosis.. |
| 71. |
KAWAMURA Wataru, KAWAMURA Wataru, MIURA Yutaka, KOKURYO Daisuke, AOKI Ichio, KANO Mitsunobu R, NISHIYAMA Nobuhiro, SAGA Tsuneo, KISHIMURA Akihiro, KATAOKA Kazunori, KATAOKA Kazunori, SPIO‐loaded and cyclic RGD installed polyion complex vesicles for the neovascular imaging of orthotopic glioblastomas, JSMI Report, 9, 2, 152, 2016.04. |
| 72. |
Takahiro Nomoto, Peng Mi, Kazuko Toh, Yu Matsumoto, Yuji Morimoto, Akihiro Kishimura, Nobuhiro Nishiyama, Kazunori Kataoka, Calcium phosphate-based organic-inorganic hybrid nanocarriers with pH-responsive on/off switch for photodynamic therapy, BIOMATERIALS SCIENCE, 10.1039/c6bm00011h, 4, 5, 826-838, 2016.03. |
| 73. |
Chan Woo Kim, Daisuke Asai, Jeong-Hun Kang, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Reversal of efflux of an anticancer drug in human drug-resistant breast cancer cells by inhibition of protein kinase C alpha (PKC alpha) activity, TUMOR BIOLOGY, 10.1007/s13277-015-3963-4, 37, 2, 1901-1908, 2016.02, P-glycoprotein (Pgp) is a 170-kDa transmembrane protein that mediates the efflux of anticancer drugs from cells. Pgp overexpression has a distinct role in cells exhibiting multidrug resistance (MDR). We examined reversal of drug resistance in human MDR breast cancer cells by inhibition of protein kinase C alpha (PKC alpha) activity, which is associated with Pgp-mediated efflux of anticancer drugs. PKC alpha activity was confirmed by measurement of phosphorylation levels of a PKC alpha-specific peptide substrate (FKKQGSFAKKK-NH2), showing relatively higher basal activity in drug-resistant MCF-7/ADR cells (84 %) than that in drug-sensitive MCF-7 cells (63 %). PKC alpha activity was effectively suppressed by the PKC inhibitor, Ro-31-7549, and reversal of intracellular accumulation of doxorubicin was observed by inhibition of PKC alpha activity in MCF-7/ADR cells compared with their intrinsic drug resistance. Importantly, increased accumulation of doxorubicin could enhance the therapeutic efficacy of doxorubicin in MDR cells significantly. These results suggest a potential for overcoming MDR via inhibition of PKC alpha activity with conventional anticancer drugs.. |
| 74. |
Md Zahangir Hosain, Takeshi Mori, Akihiro Kishimura, Yoshiki Katayama, Synergy between phenotypic modulation and ROS neutralization in reduction of inflammatory response of hypoxic microglia by using phosphatidylserine and antioxidant containing liposomes, JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION, 10.1080/09205063.2015.1125565, 27, 3, 290-302, 2016.02, Neuroinflammation caused by microglial activation is a key contributing factor in neurological disorders such as those involving ischaemia. Excess production of reactive oxygen species (ROS) and nitric oxide (NO) stimulates the inflammatory response during ischaemia, significantly damaging cells. Inhibition of inflammatory activation of microglia is a promising potential treatment approach for neurological diseases. In this study, we introduce -tocopherol and phosphatidylserine (PS) containing liposomes (PST-liposomes) to inhibit the microglial inflammatory response. PS is known to have anti-inflammatory effects on microglia by modulating the microglial phenotype, while -tocopherol is an antioxidant, known to neutralize ROS. We found that both PS-containing liposomes (PS-liposomes) and PST-liposomes, as compared with phosphatidylcholine containing liposomes, significantly increased viability of hypoxia-treated microglia. The PST-liposomes functioned better than the PS-liposomes and we attribute this superior effect to a synergy between PS and -tocopherol. This synergic action of PST-liposomes was illustrated in their ability, when incubated with microglia, to reduce NO and pro-inflammatory cytokine (TNF-) production and increase anti-inflammatory cytokine (TGF-1) production. Thus, the improved viability of hypoxia-treated microglia when treated with PST-liposomes involved anti-inflammatory effects, including ROS neutralization, as well as induction of a microglial phenotypic change. Our results suggest that PST-liposomes represent a potential therapeutic approach to reducing ischaemic injury in the brain.. |
| 75. |
Yasutaka Anraku, Akihiro Kishimura, Mako Kamiya, Sayaka Tanaka, Takahiro Nomoto, Kazuko Toh, Yu Matsumoto, Shigeto Fukushima, Daiki Sueyoshi, Mitsunobu R. Kano, Yasuteru Urano, Nobuhiro Nishiyama, Kazunori Kataoka, Systemically Injectable Enzyme-Loaded Polyion Complex Vesicles as In Vivo Nanoreactors Functioning in Tumors, ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 10.1002/anie.201508339, 55, 2, 560-565, 2016.01, The design and construction of nanoreactors are important for biomedical applications of enzymes, but lipid-and polymeric-vesicle-based nanoreactors have some practical limitations. We have succeeded in preparing enzyme-loaded polyion complex vesicles (PICsomes) through a facile protein-loading method. The preservation of enzyme activity was confirmed even after cross-linking of the PICsomes. The crosslinked beta-galactosidase-loaded PICsomes (beta-gal@PICsomes) selectively accumulated in the tumor tissue of mice. Moreover, a model prodrug, HMDER-beta Gal, was successfully converted into a highly fluorescent product, HMDER, at the tumor site, even 4 days after administration of the beta-gal@PICsomes. Intravital confocal microscopy showed continuous production of HMDER and its distribution throughout the tumor tissues. Thus, enzyme-loaded PICsomes are useful for prodrug activation at the tumor site and could be a versatile platform for enzyme delivery in enzyme prodrug therapy.. |
| 76. |
Akihiro Kishimura, An emerging material “PICsome”
A hot zone between “PEG” and “PEG”, Drug Delivery System, 10.2745/dds.31.308, 31, 4, 308-319, 2016.01, Recently, nanomaterials constructed by molecular self-assembly have gathered much attention to develop nano-devices incorporated with many types of drugs. Particularly, hollow capsules are one of promising materials, and recently, we have developed polyion complex vesicles, PICsomes, as novel polymeric vesicles. The most advantageous feature of PICsomes is its simple preparation process: Typically, they can be prepared by simple mixing of oppositely charged block copolymers consisting of poly(ethylene glycol) (PEG) and charged poly(amino acid)s in an aqueous medium. Moreover, many other unique properties of PICsomes have been reported, such as facile tuning of vesicle sizes ranging from 100-400 nm while keeping monodispersed size distribution, semipermeable vesicle membrane, facile loading of various water-dispersed materials, long blood circulation after crosslinking, excellent tumor accumulation based on the enhanced permeability and retention (EPR) effect, and so on. The present review article describes basic design and synthetic strategy of PICsomes, fundamental properties of PICsomes, and recent applications of PICsomes to drug delivery system.. |
| 77. |
Y. Anraku, A. Kishimura, M. Kamiya, S. Tanaka, T. Nomoto, K. Toh, Y. Matsumoto, S. Fukushima, D. Sueyoshi, M. R. Kano, Y. Urano, N. Nishiyama, K. Kataoka, Systemically injectable enzyme-loaded polyion complex vesicles as in vivo nanoreactors functioning in tumors, Angewandte Chemie International Edition, 10.1002/ange.201508339, 128, 2, 570-575, 2016.01. |
| 78. |
Purnima Kumar, Md Zahangir Hosain, Jeong-Hun Kang, Masafumi Takeo, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Suppression of atopic dermatitis in mice model by reducing inflammation utilizing phosphatidylserine-coated biodegradable microparticles, JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION, 10.1080/09205063.2015.1100844, 26, 18, 1465-1474, 2015.12, Controlling inflammatory response is important to avoid chronic inflammation in many diseases including atopic dermatitis (AD). In this research, we tried using a phosphatidylserine (PS)-coated microparticles in the AD mouse model for achieving the modulation of the macrophage phenotype to an anti-inflammatory state. Here, we prepared poly (D,L-lactic acid) microparticle coated with PS on the outside shell. We confirmed the cellular uptake of the PS-coated microparticle, which leads to the significant downregulation of the inflammatory cytokine production. In the mouse model of AD, the PS-coated microparticle was injected subcutaneously for a period of 12days. The mice showed significant reduction in the development of AD symptoms comparing with the mice treated with the PC-coated microparticle.. |
| 79. |
Omer F. Mutaf, Akihiro Kishimura, Yuki Mochida, Ahram Kim, Kazunori Kataoka, Induction of Secondary Structure through Micellization of an Oppositely Charged Pair of Homochiral Block-and Homopolypeptides in an Aqueous Medium, MACROMOLECULAR RAPID COMMUNICATIONS, 10.1002/marc.201500368, 36, 22, 1958-1964, 2015.11, Polyion complex (PIC) formation is an attractive method for obtaining molecular assemblies owing to their facile fabrication process in aqueous media, but more insights are required in order to control the higher-dimensional structures of polypeptide-based PICs. Herein, the PIC formation behavior of oppositely charged homochiral polypeptides, poly-L-lysine and poly(ethylene glycol)-b-poly(L-glutamate) (PEG-PLG), and their secondary structures are carefully studied in water. PIC formation takes place in a polymer concentration-dependent manner, and clear beta-sheet formation is observed at polymer concentrations >= 0.3 mg mL(-1). The results also confirm that multimolecular aggregation is a prerequisite for beta-sheet formation, which indicates that the inner hydrophobic environment of PICs is favorable for beta-sheet formation. Furthermore, the PEG weight fraction, stereoregularity of the polypeptide, and ionic strength of the solutions are found to be key factors for generating a secondary structure, presumably because these factors can contribute to the tuning of the inner environment of PICs. This method of producing water-soluble nanoassemblies from oppositely charged polypeptides may expedite self-assembly studies in biological systems and be incorporated into various molecular systems to exploit protein-mimicking features.. |
| 80. |
Kai Li, Hikari Sato, Chan Woo Kim, Yuta Nakamura, Guo Xi Zhao, Daiki Funamoto, Takanobu Nobori, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Tumor accumulation of protein kinase-responsive gene carrier/DNA polyplex stabilized by alkanethiol for intravenous injection, JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION, 10.1080/09205063.2015.1054922, 26, 11, 657-668, 2015.07, We synthesized polymeric gene carriers consisting of poly-L-lysine (PLL) main chain modified both with substrate peptide for protein kinase C (PKC) and alkanethiol (pentadecanethiol). Due to the grafted substrate peptide, the polyplex prepared from these carriers is expected to show gene expression triggered by the phosphorylation of the peptide by intracellular PKC. The modified alkanethiol on the main chain stabilized the polyplex both via disulfide crosslinking and hydrophobic interaction. The polyplex found to show gene expression in vitro when the alkanethiol content in the main chain was enough low (4-mol%-modification of PLL's epsilon-amine group) to minimize cytotoxic effect. Even though the content of alkanethiol is low, the polyplex had significant stability in a model serum solution and showed longer blood circulation in vivo. The polyplex clearly accumulated in tumor after intravenous injection.. |
| 81. |
Wataru Kawamura, Yutaka Miura, Daisuke Kokuryo, Kazuko Toh, Naoki Yamada, Takahiro Nomoto, Yu Matsumoto, Daiki Sueyoshi, Xueying Liu, Ichio Aoki, Mitsunobu R. Kano, Nobuhiro Nishiyama, Tsuneo Saga, Akihiro Kishimura, Kazunori Kataoka, Density-tunable conjugation of cyclic RGD ligands with polyion complex vesicles for the neovascular imaging of orthotopic glioblastomas, SCIENCE AND TECHNOLOGY OF ADVANCED MATERIALS, 10.1088/1468-6996/16/3/035004, 16, 3, 035004, 2015.06, Introduction of ligands into 100 nm scaled hollow capsules has great potential for diagnostic and therapeutic applications in drug delivery systems. Polyethylene glycol-conjugated (PEGylated) polyion complex vesicles (PICsomes) are promising hollow nano-capsules that can survive for long periods in the blood circulation and can be used to deliver water-soluble macromolecules to target tissues. In this study, cyclic RGD (cRGD) peptide, which is specifically recognized by alpha(V)beta(3) and alpha(v)beta(5) integrins that are expressed at high levels in the neovascular system, was conjugated onto the distal end of PEG strands on PICsomes for active neovascular targeting. Density-tunable cRGD-conjugation was achieved using PICsomes with definite fraction of end-functionalized PEG, to substitute 20, 40, and 100% of PEG distal end of the PICsomes to cRGD moieties. Compared with control-PICsomes without cRGD, cRGD-PICsomes exhibited increased uptake into human umbilical vein endothelial cells. Intravital confocal laser scanning microscopy revealed that the 40%-cRGD-PICsomes accumulated mainly in the tumor neovasculature and remained in the perivascular region even after 24 h. Furthermore, we prepared superparamagnetic iron oxide (SPIO)-loaded cRGD-PICsomes for magnetic resonance imaging (MRI) and successfully visualized the neovasculature in an orthotopic glioblastoma model, which suggests that SPIO-loaded cRGD-PICsomes might be useful as a MRI contrast reagent for imaging of the tumor microenvironment, including neovascular regions that overexpress alpha(V)beta(3) integrins.. |
| 82. |
Daiki Funamoto, Daisuke Asai, Kazuki Sato, Yoko Yamaguchi, Chan Woo Kim, Hikari Sato, Elnaz Nakhaei, Shingo Matsumoto, Takuma Yoshikawa, Koichi Sasaki, Tatsuhiro Yamamoto, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Antibody Internalization into Living Cells via Cross linker-mediated Endocytosis, CHEMISTRY LETTERS, 10.1246/cl.141157, 44, 4, 468-470, 2015.04, We reported here an alternative strategy of antibody internalization by crosslinker-mediated endocytosis. This crosslinker consists of IgG binding peptide and folic acid as cancer targeting ligands toward the folate receptor, which highly expresses in many cancer cells surface. This crosslinker successfully facilitated antibody internalization into cancer cell by the folate receptor-mediated endocytosis.. |
| 83. |
Masafumi Takeo, Cuicui Li, Masayoshi Matsuda, Hiroko Nagai, Wataru Hatanaka, Tatsuhiro Yamamoto, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Optimum design of amphiphilic polymers bearing hydrophobic groups for both cell surface ligand presentation and intercellular cross-linking, JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION, 10.1080/09205063.2015.1007414, 26, 6, 353-368, 2015.04, Amphiphilic polymers bearing hydrophobic alkyl groups are expected to be applicable for both ligand presentation on the cell surface and intercellular crosslinking. To explore the optimum design for each application, we synthesized eight different acyl-modified dextrans with varying molecular weight, alkyl length, and alkyl modification degree. We found that the behenate-modified polymers retained on the cell surface longer than the palmitate-modified ones. Since the polymers were also modified with biotin, streptavidin can be presented on the cell surface through biotin-streptavidin recognition. The duration of streptavidin on the cell surface is longer in the behenate-modified polymer than the palmitate-modified one. As for the intercellular crosslinking, the palmitate-modified polymers were more efficient than the behenate-modified polymers. The findings in this research will be helpful to design the acyl-modified polymers for the cell surface engineering.. |
| 84. |
Daiki Funamoto, Daisuke Asai, Kazuki Sato, Yoko Yamaguchi, Chan Woo Kim, Hikari Sato, Elnaz Nakhaei, Shingo Matsumoto, Takuma Yoshikawa, Koichi Sasaki, Tatsuhiro Yamamoto, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Antibody Internalization into Living Cells via Cross linker-mediated Endocytosis, CHEMISTRY LETTERS, 10.1246/cl.141157, 44, 4, 468-470, 2015.04, We reported here an alternative strategy of antibody internalization by crosslinker-mediated endocytosis. This crosslinker consists of IgG binding peptide and folic acid as cancer targeting ligands toward the folate receptor, which highly expresses in many cancer cells surface. This crosslinker successfully facilitated antibody internalization into cancer cell by the folate receptor-mediated endocytosis.. |
| 85. |
腫瘍においてクスリを「つくる」酵素封入PICsomeの創製. |
| 86. |
Eun Kyung Lee, Chan Woo Kim, Hiroyuki Kawanami, Akihiro Kishimura, Takuro Niidome, Takeshi Mori, Yoshiki Katayama, Utilization of a PNA-peptide conjugate to induce a cancer protease-responsive RNAi effect, RSC Advances, 10.1039/c5ra17737e, 5, 104, 85816-85821, 2015.01, Small interfering RNA (siRNA) is regarded as a promising tool for cancer therapy because of the wide applicability to various cancer-related genes. However, non-specific delivery of siRNA is one of the major causes of adverse effects. To access the issue, here we designed a new siRNA system which turns on RNAi responding to a cancer cell-specific protease, cathepsin B. The system uses a peptide nucleic acid (PNA)-peptide conjugate to provide this protease-responsive activation. The PNA-peptides were found to form hybrids with double-stranded RNAs with complementary protruding regions, which then affected the susceptibility of dsRNA to Dicer. The dsRNA/PNA-peptide hybrids were activated in cancer cells with a high cathepsin B activity to show RNAi.. |
| 87. |
Masayoshi Matsuda, Wataru Hatanaka, Masafumi Takeo, Chan Woo Kim, Takuro Niidome, Tatsuhiro Yamamoto, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Short Peptide Motifs for Long-Lasting Anchoring to the Cell Surface, BIOCONJUGATE CHEMISTRY, 10.1021/bc500465j, 25, 12, 2134-2143, 2014.12, A rational design strategy has been developed for the construction of stable peptide-based anchors for the efficient modification of cell surfaces. Six types of peptide composed of five residues with divalent hydrophobic groups have been designed using this new strategy. Among them, a peptide with a sequence of NBD-Lys-Lys(X)-Lys-Lys-Lys(X)-NH2 (NBD: fluorophore, Lys(X): N-e-palmitoyl-l-lysine) was found to show the highest modification efficacy and longevity in culture medium. The good performance of this peptide was attributed to (1) its high aqueous solubility, which allowed it to partition from the medium to the cell surface, and (2) the high binding affinity of the saturated palmitoyl groups to the cell membrane. We found that the distribution of the peptide was affected by recycling endosome, which enabled the representation of the peptide following its endocytotic disappearance from the cell membrane. Biotin was also presented on the cell surface using this peptide-based anchor to examine its recognition by streptavidin. The efficacy of the recognition process increased as the length of the oligoethylene glycol spacer increased, indicating that it was necessary for the biotin tag to move away from the membrane glycoproteins on the cell surface to facilitate its efficient recognition by streptavidin.. |
| 88. |
Hikari Sato, Yuta Nakamura, Elnaz Nakhaei, Daiki Funamoto, Chan Woo Kim, Tatsuhiro Yamamoto, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, A Liposome Reversibly Coated with Serum Albumin, CHEMISTRY LETTERS, 10.1246/cl.140432, 43, 9, 1481-1483, 2014.09, Herein, we reported a design of liposome affording a reversible coating of serum albumin on its surface. We synthesized a lipid modified with a serum albumin-specific endogenous ligand and prepared liposome, including this lipid. We successfully confirmed the coating of the liposome surface with serum albumin. The liposome presented here would be applicable to a drug carrier with enhanced blood circulation.. |
| 89. |
Kensuke Osada, Horacio Cabral, Yuki Mochida, Yasutaka Anraku, Akihiro Kishimura, Nobuhiro Nishiyama, Kazunori Kataoka, Metallosomes as bioactive polymersomes formed by supramolecular assembly of block copolymer-metal complexation, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 248, 2014.08. |
| 90. |
D. Sueyoshi, A. Kishimura, H. Oana, Y. Anraku, M. Takai, M. Washizu, K. Kataoka, Microchannel-assisted preparation of polyion complex vesicles and real-time observation of their dynamic responses to external electric fields, MicroTAS, 3, 1882-1884, 2014.08. |
| 91. |
Development of SPIO-loaded Unilamellar Polyion Complex Vesicles (SPIO-Cy5-PICsomes) as a High Relaxivity Contrast Agent for Early-stage Tumor Detection. |
| 92. |
Sayan Chuanoi, Yasutaka Anraku, Mao Hori, Akihiro Kishimura, Kazunori Kataoka, Fabrication of Polyion Complex Vesicles with Enhanced Salt and Temperature Resistance and Their Potential Applications as Enzymatic Nanoreactors, BIOMACROMOLECULES, 10.1021/bm500127g, 15, 7, 2389-2397, 2014, 2014.07, Integrating catalytic functions into polymeric vesicles through enzyme entrapment is appealing for bioreactor fabrication, yet there are critical issues regarding the regulation of solute transport through membranes and enzyme loading without denaturation. Polyion complex vesicles (PICsomes) with semipermeable membranes and the propensity to form in water can overcome these issues; however, cross-linking is required for sufficient physiological stability. Herein, we report the first successful fabrication of non-cross-linked PICsomes with sufficient stability at physiological salinity and temperature by tuning the hydrophobicity of the aliphatic side chains in the pendant group of the constituent polyelectrolytes. Dynamic light scattering and transmission electron microscopy revealed that the intervesicular fusion and disintegration of the PICsomes was prevented and a narrow distribution was maintained at physiological salinity and temperatures. Furthermore, their application as enzymatic nanoreactors was verified even in the presence of proteases. As such, the potential utility of the PICsomes in biomedical fields was established.. |
| 93. |
Horacio Cabral, Kanjiro Miyata, Akihiro Kishimura, Nanodevices for studying nano-pathophysiology, ADVANCED DRUG DELIVERY REVIEWS, 10.1016/j.addr.2014.06.003, 74, 35-52, 2014.07, Nano-scaled devices are a promising platform for specific detection of pathological targets, facilitating the analysis of biological tissues in real-time, while improving the diagnostic approaches and the efficacy of therapies. Herein, we review nanodevice approaches, including liposomes, nanoparticles and polymeric nanoassemblies, such as polymeric micelles and vesicles, which can precisely control their structure and functions for specifically interacting with cells and tissues. These systems have been successfully used for the selective delivery of reporter and therapeutic agents to specific tissues with controlled cellular and subcellular targeting of biomolecules and programmed operation inside the body, suggesting a high potential for developing the analysis for nano-pathophysiology. (C) 2014 Elsevier B.V. All rights reserved.. |
| 94. |
Takanobu Nobori, Shujiro Shiosaki, Takeshi Mori, Riki Toita, Chan Woo Kim, Yuta Nakamura, Akihiro Kishimura, Takuro Niidome, Yoshiki Katayama, Fluorescent Polyion Complex Nanoparticle That Incorporates an Internal Standard for Quantitative Analysis of Protein Kinase Activity, BIOCONJUGATE CHEMISTRY, 10.1021/bc500142j, 25, 5, 869-872, 2014.05, We demonstrate a polyion complex (PIC) nanoparticle that contains both a responsive fluorophore and an "internal standard" fluorophore for quantitative measurement of protein kinase (PK) activity. The PK-responsive fluorophore becomes more fluorescent with PK-catalyzed phosphorylation of substrate peptides incorporated in the PIC, while fluorescence from the internal standard remains unchanged during phosphorylation. This new concept will be useful for quantitative PK assays and the discovery of PK inhibitors.. |
| 95. |
Arie Wibowo, Kensuke Osada, Hiroyuki Matsuda, Yasutaka Anraku, Haruko Hirose, Akihiro Kishimura, Kazunori Kataoka, Morphology Control in Water of Polyion Complex Nanoarchitectures of Double-Hydrophilic Charged Block Copolymers through Composition Tuning and Thermal Treatment, MACROMOLECULES, 10.1021/ma500314d, 47, 9, 3086-3092, 2014.05, Polyion complexes (PICs) are attractive as eco-friendly materials, because they offer simple and fast preparation methods to exert various functionalities in aqueous medium. However, control of nanoarchitectures in PIC materials has not been fully realized, except for the case of micelles and unilamellar vesicles formed from block ionomers. Here, the procedure to control PIC nanoarchitectures with various morphologies was established for the first time by careful tuning in the composition of PICs made from PEG-based block-ionomers with a varying amount of homoionomers as additive to modulate the PEG weight fraction (f(PEG)) in the obtained PICs. Accordingly, the variation in f(PEG) from 12.1% to 6.5% induced vigorous transition in the microphase separated structures of PICs basically from micelle to lamella via cylindrical network. Notably, uniformed lamella with alternative layers of PEG and PIG domains was found at elevated temperature (70 degrees C), which, by lowering temperature, reversibly transformed to cylindrical P1C network apparently with connected aqueous channel in mesoscopic scale.. |
| 96. |
KAWAMURA Wataru, MIURA Yutaka, KOKURYO Daisuke, AOKI Ichio, KISHIMURA Akihiro, KATAOKA Kazunori, Cyclic RGD‐linked Polyion Complex Vesicles (PICsomes) for Neovascular Targeting and Glioblastoma Imaging, 高分子学会予稿集(CD-ROM), 63, 1, ROMBUNNO.3PC125, 2014.05. |
| 97. |
Akihiro Kishimura, Morphology Control in Water of Polyion Complex Nanoarchitectures of Double-Hydrophilic Charged Block Copolymers through Composition Tuning and Thermal Treatment. , American Chemical Society, 10.1021/ma500314d , 47, 9, 3086-3092, 2014, 2014.04. |
| 98. |
Daisuke Kokuryo, Yasutaka Anraku, Akihiro Kishimura, Sayaka Tanaka, Mitsunobu R. Kano, Jeff Kershaw, Nobuhiro Nishiyama, Tsuneo Saga, Ichio Aoki, Kazunori Kataoka, SPIO-PICsome: Development of a highly sensitive and stealth-capable MRI nano-agent for tumor detection using SPIO-loaded unilamellar polyion complex vesicles (PICsomes) (vol 169, pg 220, 2013), JOURNAL OF CONTROLLED RELEASE, 10.1016/j.jconrel.2014.01.021, 178, 125-125, 2014.03. |
| 99. |
Guo Xi Zhao, Hiroyuki Tanaka, Chan Woo Kim, Kai Li, Daiki Funamoto, Takanobu Nobori, Yuta Nakamura, Takuro Niidome, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Histidinylated poly-L-lysine-based vectors for cancer-specific gene expression via enhancing the endosomal escape, JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION, 10.1080/09205063.2013.879562, 25, 5, 519-534, 2014.03, In this work, we synthesized a series of poly-L-lysine (PLL)-based polymers for gene delivery, by modifying the PLL with both cationic peptide and histidine. The peptide moieties serve as cationic centers for polyplex formation, and also as substrates for protein kinase C alpha (PKC alpha), which is specifically activated in many types of cancer cells, to achieve cancer-specific gene expression. The histidine groups serve as buffering moieties to increase the ability of the plasmid DNA (pDNA)-polymer complex (polyplex) to escape the endosome and thus to promote expression of the pDNA in the transfected cells. The facile synthesis of the polymers proceeded by modifying the PLL with side-group-protected peptide and protected histidine, followed by deprotection of the functional groups. The synthesized polymers showed significant buffering capacity over the neutral to acidic pH range and showed less cytotoxicity in vitro compared with histidine-unmodified polymers. The polyplexes successfully showed PKC alpha-responsive gene expression immediately after their introduction into cancer cells and the gene expression continued for at least 24 h. These PLL-based carriers thus show promise for cancer-targeted gene therapy.. |
| 100. |
Kyohei Tobinaga, Cuicui Li, Masafumi Takeo, Masayoshi Matsuda, Hiroko Nagai, Takuro Niidome, Tatsuhiro Yamamoto, Akihiro Kishimura, Takeshi Mori, Yoshiki Katayama, Rapid and serum-insensitive endocytotic delivery of proteins using biotinylated polymers attached via multivalent hydrophobic anchors, JOURNAL OF CONTROLLED RELEASE, 10.1016/j.jconrel.2013.12.024, 177, 27-33, 2014.03, We have designed biotinylated polymers as synthetic receptors that have multiple alkyl groups for endocytotic delivery of target proteins. The polymers were stably attached to a cell surface via multivalent anchoring. The presented biotin was bound to streptavidin (SA) on the cell surface, and, via an endocytotic pathway, the cell rapidly internalized the biotinylated polymer/SA complex. The cell's uptake of the complex was not inhibited by the presence of 10% fetal bovine serum, and its efficacy for the uptake of SA was the highest when compared with commercial reagents and single-anchored-type synthetic receptors. The synthetic receptor-mediated endocytosis can be used generally for other kind of protein by using SA as an adaptor molecule between a target protein and the cell-surface presented biotin. (C) 2013 Elsevier B.V. All rights reserved.. |
| 101. |
Christopher V. Synatschke, Takahiro Nomoto, Horacio Cabral, Melanie Foertsch, Kazuko Toh, Yu Matsumoto, Kozo Miyazaki, Andreas Hanisch, Felix H. Schacher, Akihiro Kishimura, Nobuhiro Nishiyama, Axel H. E. Mueller, Kazunori Kataoka, Multicompartment Micelles with Adjustable Poly(ethylene glycol) Shell for Efficient in Vivo Photodynamic Therapy, ACS NANO, 10.1021/nn4028294, 8, 2, 1161-1172, 2014.02, We describe the preparation of well-defined multicompartment micelles from polybutadiene-block-poly(1-methyl-2-vinyl pyridinium methyl sulfate)-block-poly(methacrylic acid) (BVqMAA) triblock terpolymers and their use as advanced drug delivery systems for photodynamic therapy (PDT). A porphyrazine derivative was incorporated into the hydrophobic core during self-assembly and served as a model drug and fluorescent probe at the same time. The initial micellar corona is formed by negatively charged PMAA and could be gradually changed to poly(ethylene glycol) (PEG) in a controlled fashion through interpolyelectrolyte complex formation of PMAA with positively charged poly(ethylene glycol)-block-poly(L-lysine) (PLL-b-PEG) diblock copolymers. At high degrees of PEGylation, a compartmentalized micellar corona was observed, with a stable bottlebrush-on-sphere morphology as demonstrated by cryo-TEM measurements. By in vitro cellular experiments, we confirmed that the porphyrazine-loaded micelles were PDT-active against A549 cells. The corona composition strongly influenced their in vitro PDT activity, which decreased with increasing PEGylation, correlating with the cellular uptake of the micelles. Also, a PEGylation-dependent influence on the in vivo blood circulation and tumor accumulation was found. Fully PEGylated micelles were detected for up to 24 h in the bloodstream and accumulated in solid subcutaneous A549 tumors, while non- or only partially PEGylated micelles were rapidly cleared and did not accumulate in tumor tissue. Efficient tumor growth suppression was shown for fully PEGylated micelles up to 20 days, demonstrating PDT efficacy in vivo.. |
| 102. |
Sayan Chuanoi, Akihiro Kishimura, Wen-Fei Dong, Yasutaka Anraku, Yuichi Yamasaki, Kazunori Kataoka, Structural factors directing nanosized polyion complex vesicles (Nano-PICsomes) to form a pair of block aniomer/homo catiomers: studies on the aniomer segment length and the catiomer side-chain structure, POLYMER JOURNAL, 10.1038/pj.2013.82, 46, 2, 130-135, 2014.02, Much attention has been devoted to precise control of the size and morphology in nanosized molecular assemblies for a wide range of materials applications. Recently, we reported observing submicron/nanosized polyion complex vesicles (Nano-PICsomes) with a narrow size distribution, synthesized using specific types of homocatiomers and polyethylene glycol (PEG)-based block aniomers. However, only one example of Nano-PICsomes has been reported to date. Here, the role of the chemical composition of PEG-based block aniomers and the chemical structures of the side chains of homocatiomers were carefully examined to better understand the formation of Nano-PICsomes. Transmission electron microscopy and dynamic light scattering analyses of Nano-PICsomes revealed that a longer length of ionic segments in the block aniomers or a PEG weight fraction (f(PEG)) 10% produced spherical micelles. In addition, the homocatiomers containing longer aliphatic side chains (e. g., five or six carbon atoms) favored the formation of Nano-PICsomes, whereas those containing shorter aliphatic side chains produced irregularly shaped PIC micelles. Accordingly, f(PEG) and the length of the side chain were found to be the key factors that control the morphologies of Nano-PICsomes. Insights gained from this study can broaden the spectrum of the design of Nano-PICsomes for use in a diverse range of material applications.. |
| 103. |
Huabing Chen, Ling Xiao, Yasutaka Anraku, Peng Mi, Xueying Liu, Horacio Cabral, Aki Inoue, Takahiro Nomoto, Akihiro Kishimura, Nobuhiro Nishiyama, Kazunori Kataoka, Polyion Complex Vesicles for Photoinduced Intracellular Delivery of Amphiphilic Photosensitizer, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 10.1021/ja406992w, 136, 1, 157-163, 2014, 2014.01, Polymer vesicles formed by a pair of oppositely charged poly(ethylene glycol) (PEG)-based block aniomer and homocatiomer, termed "PICsomes", have tunable size, and are characterized by unique semipermeable property due to the flexible and tunable hydrophilicity of polyion complex (PIC) membranes. The PICsomes can encapsulate a variety of molecules in an inner aqueous phase just by a simple vortex mixing of solution, expecting their utility as nanocontainers of substances with biomedical interests. Here, we report on a new functionality of the PICsomes: photoinduced release of photoactive agents for intracellular drug delivery. A potent photosensitizer, Al(III) phthalocyanine chloride disulfonic acid (AlPcS2a), was efficiently incorporated into the PICsomes (11%(w/w)), and its quick release was induced by photoirracliation possibly due to the photochemical damage of the PIG membranes. The combination of a high-resolution fluorescent confocal microscopy and a lysosome membrane-specific staining method revealed that such photoinduced release of AlPcS2a occurred even in the lysosomes of living cells after endocytic internalization. Simultaneously, the released AlPcS2a photochemically affected the integrity of the lysosomal membranes, leading to the translocation of AlPcS2a and PICsomes themselves to the cytoplasm. Consequently, the AlPcS2a-encapsulated PICsomes (AIPcS2a-PICsomes) exhibited appreciably stronger photocytotoxicity compared with free AlPcS2a alone. Thus, the AlPcS2a-PICsomes have promising feasibility for the photodynamic therapy or the photoinduced cytoplasmic delivery of therapeutic molecules.. |
| 104. |
Peng Mi, Daisuke Kokuryo, Horacio Cabral, Michiaki Kumagai, Takahiro Nomoto, Ichio Aoki, Yasuko Terada, Akihiro Kishimura, Nobuhiro Nishiyama, Kazunori Kataoka, Hydrothermally synthesized PEGylated calcium phosphate nanoparticles incorporating Gd-DTPA for contrast enhanced MRI diagnosis of solid tumors, JOURNAL OF CONTROLLED RELEASE, 10.1016/j.jconrel.2013.10.038, 174, 63-71, 2014.01, Organic-inorganic hybrid nanoparticles with calciumphosphate (CaP) core and PEGylated shell were developed to incorporatemagnetic resonance imaging (MRI) contrast agent diethylenetriaminepentaacetic acid gadolinium (III) (Gd-DTPA) for noninvasive diagnosis of solid tumors. A two-step preparation method was applied to elaborate hybrid nanoparticleswith a z-average hydrodynamic diameter about 80 nm, neutral surface xi-potential and high colloidal stability in physiological environments by self-assembly of poly(ethylene glycol)-b-poly(aspartic acid) block copolymer, Gd-DTPA, and CaP in aqueous solution, followed with hydrothermal treatment. Incorporation into the hybrid nanoparticles allowed Gd-DTPA to show significant enhanced retention ratio in blood circulation, leading to high accumulation in tumor positions due to enhanced permeability and retention (EPR) effect. Moreover, Gd-DTPA revealed above 6 times increase of relaxivity in the nanoparticle system compared to free form, and eventually, selective and elevated contrast enhancements in the tumor positions were observed. These results indicate the high potential of Gd-DTPA-loaded PEGylated CaP nanoparticles as a novel contrast agent for noninvasive cancer diagnosis. (C) 2013 Elsevier B.V. All rights reserved.. |
| 105. |
D. Sueyoshi, Akihiro Kishimura, H. Oana, Y. Anraku, M. Takai, M. Washizu, K. Kataoka, Microchannel-assisted preparation of polyion complex vesicles and real-time observation of their dynamic responses to external electric fields, 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2014
18th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2014, 1882-1884, 2014, Polyion complex (PIC) vesicles, PICsomes, are polymeric vesicles which can form through electrostatic interaction between an oppositely charged polyethylene glycol (PEG)-based block ionomers and homo ionomers. In this study, for further utilization and understanding of their characteristic dynamic properties in aqueous media, we conducted real-time observation of their responses to external electric fields (EFs). This observation was facilitated by a newly designed microfluidic channel, which enabled preparation of micrometer-sized PICsomes and observation of their behaviors under controlled EF conditions. Their non-trivial responses and their EF dependency were carefully evaluated, and some dynamic behaviors were observed in several EF conditions.. |
| 106. |
Akihiro Kishimura, Development of polyion complex vesicles (PICsomes) from block copolymers for biomedical applications, POLYMER JOURNAL, 10.1038/pj.2013.33, 45, 9, 892-897, 2013.09, Polyion complex (PIC) formation is one of the most powerful techniques for obtaining molecular self-assemblies in aqueous media. The simple preparation process based on multiple electrostatic interactions is quite attractive for material syntheses, as well as biomedical applications. Therefore, it is desirable to control PIC architectures at the nanoscale in order to expand the scope of PIC materials. In this review article, recent progress on PIC vesicles (PICsomes) is summarized. PICsomes were first developed by my research group, and we recently succeeded in controlling the sizes and structural uniformity of the vesicles. Furthermore, the characteristic dynamic nature of PICs was revealed: PICs were found to exhibit reversible association/dissociation and structural transformation. We demonstrated that crosslinking the PIC layers of PICsomes is a powerful method for tuning properties such as stability and permeability. Finally, the potential utility of PICsomes for drug delivery nanocarriers was examined, and their future biomedical application is discussed.. |
| 107. |
Akihiro Kishimura, Development of polyion complex vesicles (PICsomes) from block copolymers for biomedical applications, Polymer Journal, 10.1038/pj.2013.33, 45, 9, 892-897, 2013.09, Polyion complex (PIC) formation is one of the most powerful techniques for obtaining molecular self-assemblies in aqueous media. The simple preparation process based on multiple electrostatic interactions is quite attractive for material syntheses, as well as biomedical applications. Therefore, it is desirable to control PIC architectures at the nanoscale in order to expand the scope of PIC materials. In this review article, recent progress on PIC vesicles (PICsomes) is summarized. PICsomes were first developed by my research group, and we recently succeeded in controlling the sizes and structural uniformity of the vesicles. Furthermore, the characteristic dynamic nature of PICs was revealed: PICs were found to exhibit reversible association/dissociation and structural transformation. We demonstrated that crosslinking the PIC layers of PICsomes is a powerful method for tuning properties such as stability and permeability. Finally, the potential utility of PICsomes for drug delivery nanocarriers was examined, and their future biomedical application is discussed. © 2013 The Society of Polymer Science, Japan (SPSJ) All rights reserved.. |
| 108. |
Daisuke Kokuryo, Yasutaka Anraku, Akihiro Kishimura, Sayaka Tanaka, Mitsunobu R. Kano, Jeff Kershaw, Nobuhiro Nishiyama, Tsuneo Saga, Ichio Aoki, Kazunori Kataoka, SPIO-PICsome: Development of a highly sensitive and stealth-capable MRI nano-agent for tumor detection using SPIO-loaded unilamellar polyion complex vesicles (PICsomes), JOURNAL OF CONTROLLED RELEASE, 10.1016/j.jconrel.2013.03.016, 169, 3, 220-227, 2013.08, Size controllable polyion complex vesicles (PICsomes), composed of biocompatible poly(ethylene glycol) (PEG) and poly(amino acid) s, have an extremely prolonged lifetime in the bloodstream that enables them to accumulate effectively in tumors via the enhanced permeability and retention (EPR) effect. The purpose of this study was to use PICsomes to synthesize a highly sensitive MRI contrast agent for more precise tumor detection. We synthesized SPIO-Cy5-PICsomes (superparamagnetic iron oxide nanoparticle-loaded Cy5-cross-linked Nano-PICsomes) and characterized them using dynamic light scattering and transmission electron microscopy in vitro and evaluated their ability to detect subcutaneously grafted tumors in vivo with MRI. The transverse relaxivity (r(2)) of the SPIO-Cy5-PICsomes (r(2) = 663 +/- 28 mM(-1) s(-1)) was 2.54 times higher than that of bare clinically-used SPIO. In in vivo MRI experiments on mice subcutaneously grafted with colon-26 tumor cells, the tumor signal was significantly altered at 3 h after SPIO-Cy5-PICsome administration and persisted for at least 24 h. Small and early-stage in vivo tumors (3 days after grafting, approximately 4 mm(3)) were also clearly detected with MRI. SPIO-loaded PICsomes are sensitive MRI contrast agents that can act as a powerful nanocarrier to detect small tumors for early diagnosis. (C) 2013 Elsevier B. V. All rights reserved.. |
| 109. |
N. Sasaki, M. Tatanou, Y. Anraku, A. Kishimura, K. Kataoka, K. Sato, Characterization of nanoparticle permeability on a membrane-integrated microfluidic device, MicroTAS, 3, 1818-1820, 2013.08. |
| 110. |
Hidehiro Oana, Mutsuki Morinaga, Akihiro Kishimura, Kazunori Kataoka, Masao Washizu, Direct formation of giant unilamellar vesicles from microparticles of polyion complexes and investigation of their properties using a microfluidic chamber, Soft Matter, 10.1039/c3sm00089c, 9, 22, 5448-5458, 2013.06, Although hollow microscopic capsules have a variety of potential biomedical applications, reports of organic-solvent-free methods for their preparation are rather limited. Herein, a novel approach is demonstrated for organic-solvent-free preparation of giant unilamellar vesicles utilizing the unique response of polyion complexes (PICs) to changes in additive salt concentration. A microfluidic device consisting of a main channel bearing side pockets that work as microscale reaction chambers is designed for facilitating the preparation process under an optical microscope. With this device, real-time observation of morphological transformation of individual PIC microparticles is carried out during rapid reduction of the additive salt concentration and direct formation of giant vesicles from PIC microparticles is shown. There is a quasilinear relationship between the surface areas of the formed vesicles and the volumes of the PIC microparticles, and the thickness of the vesicle membrane estimated by the relationship is indicative of the formation of a uniform unilamellar structure of the PIC membrane. Furthermore, detailed properties of the formed PIC vesicles with regard to salt response, loading of guest molecules, and permeability of the PIC membrane with/without modification of the PIC membrane by cross-linking are investigated using the microfluidic chamber. Thus, the usefulness of the microfluidic chamber for visualization and investigation of dynamic responses of microscale soft materials during changes in surrounding conditions is also demonstrated.. |
| 111. |
Development of Nano-Sized Polyion Complex Vesicle "PICsomes" and its Application to Bioimaging and Nano-Physiology. |
| 112. |
汎用性の高い酵素プロドラッグ療法用キャリアを指向した血中滞留型中空ナノ粒子(PICsome)の構築. |
| 113. |
PIC型ベシクルの特徴を活かした酵素プロドラッグ療法用キャリアの構築. |
| 114. |
超常磁性酸化鉄微粒子を内包したポリイオンコンプレックス型中空粒子SPIO‐PICsomeの開発と緩和能・腫瘍集積性評価. |
| 115. |
D. Kokuryo, Y. Anraku, A. Kishimura, M. R Kano, S. Tanaka, T. Saga, I. Aoki, K. Kataoka, SPIO-loaded unilamellar polyion complex vesicles (SPIO-Cy5-PICsomes) as a high relaxivity contrast agent for tumor detection, Proc. 21th ISMRM Scientific Meeting and Exhibition, 1869-1869, 2013.04. |
| 116. |
A. Kasuya, S. Hirakawa, K. Liu, A. Kishimura, Y. Anraku, K. Kataoka, M. Ogawa, S. Mori, H. K. Takahashi, J. Sakabe, Y. Tomono, M. Nishibori, Y. Tokura, Anti-HMGB1 antibody attenuates vascular hyperpermeability and promotes wound healing during ischemia-reperfusion injury model in mouse skin, Journal of Dermatological Science, 10.1016/j.jdermsci.2012.11.481, 69, 2, e59, 2013.02. |
| 117. |
N. Sasaki, M. Tatanou, Y. Anraku, Akihiro Kishimura, K. Kataoka, K. Sato, Characterization of nanoparticle permeability on a membrane-integrated microfluidic device, 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013
17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013, 3, 1818-1820, 2013.01, We present a microfluidic platform to characterize nanoparticle permeability, which determines the efficiency of nanoparticle-based drug delivery to tumors. We integrated porous membranes that mimic tumor vascular walls into microfluidic devices. Permeation of the nanoparticles through the membranes was studied under control of transmembrane flow, which corresponds to outward flow from blood vessels to tumors in vivo. The obtained results agreed with a theoretical model considering the effect of particle/pore size ratio on the permeation. These results indicate that the device can be a simple model to estimate the permeability of the nanoparticles.. |
| 118. |
Yasutaka Anraku, Akihiro Kishimura, Yuichi Yamasaki, Kazunori Kataoka, Living Unimodal Growth of Polyion Complex Vesicles via Two-Dimensional Supramolecular Polymerization, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 10.1021/ja3096587, 135, 4, 1423-1429, 2013.01, Understanding the dynamic behavior of molecular self-assemblies with higher-dimensional structures remains a key challenge to obtaining well-controlled and monodispersed structures. Nonetheless, there exist few systems capable of realizing the mechanism of supramolecular polymerization at higher dimensions. Herein, we report the unique self-assembling behavior of polyion complexes (PICs) consisting of poly(ethylene glycol)-polyelectrolyte block copolymer as an example of two-dimensional supramolecular living polymerization. Monodispersed and submicrometer unilamellar PIC vesicles (nano-PICsomes) displayed time-dependent growth while maintaining a narrow size distribution and a unilamellar structure. Detailed analysis of the system revealed that vesicle growth proceeded through the consumption of unit PICs (uPICs) composed of a single polycation/polyanion pair and was able to restart upon the further addition of isolated uPICs. Interestingly, the resulting vesicles underwent dissociation into uPICs in response to mechanical stress. These results clearly frame the growth as a two-dimensional supramolecular living polymerization of uPICs.. |
| 119. |
Noha Gouda, Kanjiro Miyata, R. James Christie, Tomoya Suma, Akihiro Kishimura, Shigeto Fukushima, Takahiro Nomoto, Xueying Liu, Nobuhiro Nishiyama, Kazunori Kataoka, Silica nanogelling of environment-responsive PEGylated polyplexes for enhanced stability and intracellular delivery of siRNA, BIOMATERIALS, 10.1016/j.biomaterials.2012.09.077, 34, 2, 562-570, 2013.01, In this study, poly(ethylene glycol) (PEG)-block-polycation/siRNA complexes (PEGylated polyplexes) were wrapped with a hydrated silica, termed "silica nanogelling", in order to enhance their stability and functionality. Silica nanogelling was achieved by polycondensation of soluble silicates onto the surface of PEGylated polyplexes comprising a disulfide cross-linked core. Formation of silica nanogel layer on the PEGylated cross-linked polyplexes was confirmed by particle size increase, surface charge reduction, and elemental analysis of transmission electron micrographs. Silica nanogelling substantially improved polyplex stability against counter polyanion-induced dissociation under non-reductive condition, without compromising the reductive environment-responsive siRNA release triggered by disulfide cleavage. Silica nanogelling significantly enhanced the sequence-specific gene silencing activity of the polyplexes in HeLa cells without associated cytotoxicity, probably due lower endosomal entrapment (or lysosomal degradation) of delivered siRNA. The lower endosomal entrapment of the silica nanogel system could be explained by an accelerated endosomal escape triggered by deprotonated silanol groups in the silica (the proton sponge hypothesis) and/or a modulated intracellular trafficking, possibly via macropinocytosis, as evidenced by the cellular uptake inhibition assay. Henceforth, silica nanogelling of PEGylated siRNA polyplexes is a promising strategy for preparation of stable and functional siRNA delivery vehicles. (C) 2012 Elsevier Ltd. All rights reserved.. |
| 120. |
Yuta Maeyoshi, Akinori Saeki, Shotaro Suwa, Masaaki Omichi, Hiromi Marui, Atsushi Asano, Satoshi Tsukuda, Masaki Sugimoto, Akihiro Kishimura, Kazunori Kataoka, Shu Seki, Fullerene nanowires as a versatile platform for organic electronics, Scientific reports, 10.1038/srep00600, 2, 2012.09, The development of organic semiconducting nanowires that act as charge carrier transport pathways in flexible and lightweight nanoelectronics is a major scientific challenge. We report on the fabrication of fullerene nanowires that is universally applicable to its derivatives (pristine C 60, methanofullerenes of C 61 and C 71, and indene C 60 bis-adduct), realized by the single particle nanofabrication technique (SPNT). Nanowires with radii of 811 nmwere formed via a chain polymerization reaction induced by a high-energy ion beam. Fabrication of a poly(3-hexylthiophene) (P3HT): [6,6]-phenyl C 61 butyric acid methyl ester (PC 61BM) bulk heterojunction organic photovoltaic cell including PC 61BM nanowires with precisely-controlled length and density demonstrates how application of this methodology can improve the power conversion efficiency of these inverted cells. The proposed technique provides a versatile platform for the fabrication of continuous and uniform n-type fullerene nanowires towards a wide range of organic electronics applications.. |
| 121. |
Kensuke Osada, Horacio Cabral, Yuki Mochida, Sangeun Lee, Kazuya Nagata, Tetsuya Matsuura, Megumi Yamamoto, Yasutaka Anraku, Akihiro Kishimura, Nobuhiro Nishiyama, Kazunori Kataoka, Bioactive Polymeric Metallosomes Self-Assembled through Block Copolymer-Metal Complexation, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 10.1021/ja304615y, 134, 32, 13172-13175, 2012.08, Spontaneous formation of polymeric metallosomes with uniform size (similar to 100 nm) was found to occur in aqueous medium through the reaction of an anticancer agent, (1,2-diaminocyclohexane)platinum(II) (DACHPt), with a Y-shaped block copolymer of omega-cholesteroyl-poly(L-glutamic acid) and two-armed poly(ethylene glycol) (PEGasus-PLGA-Chole). Circular dichroism spectrum measurements revealed that the PLGA segment forms an alpha-helix structure within the metallosomes, suggesting that secondary-structure formation of metallocomplexed PLGA segment may drive the self-assembly of the system into vesicular structure. These metallosomes can encapsulate water-soluble fluorescent macromolecules into their inner aqueous phase and eventually deliver them selectively into tumor tissues in mice, owing to the prolonged blood circulation. Accordingly, fluorescent imaging of the tumor was successfully demonstrated along with an appreciable antitumor activity by DACHPt moieties retained in the vesicular wall of the metallosomes, indicating the potential of metallosomes as multifunctional drug carriers.. |
| 122. |
Shuhei Murayama, Baowei Su, Kohki Okabe, Akihiro Kishimura, Kensuke Osada, Masayuki Miura, Takashi Funatsu, Kazunori Kataoka, Masaru Kato, NanoPARCEL
A method for controlling cellular behavior with external light, Chemical Communications, 10.1039/c2cc32718j, 48, 67, 8380-8382, 2012.08, We developed a simple preparation procedure for the protein encapsulated nanoparticle and used the nanoparticle for spatiotemporal activity control of various proteins. We succeeded in the local protein activation within cells by light using the nanoparticle.. |
| 123. |
Christopher V. Synatschke, Takahiro Nomoto, Akihiro Kishimura, Axel H. E. Muller, Kazunori Kataoka, Towards multifunctional drug delivery systems from ionic triblock terpolymer micelles, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 244, 2012.08. |
| 124. |
In vivoナノリアクターへの展開を指向した酵素封入PICsomeの機能評価. |
| 125. |
粒径の制御された高分子ベシクルPICsomeの体内動態と生体内イメージング応用. |
| 126. |
超常磁性酸化鉄微粒子を内包したポリイオンコンプレックス型中空ナノ粒子PICsomeを用いたin vivo MRイメージング. |
| 127. |
架橋率・サイズの異なるポリイオンコンプレックス型中空粒子(Nano‐PICsome)の基礎物性評価および体内動態評価. |
| 128. |
インテリジェントDDSキャリアを指向したポリイオンコンプレックス型中空粒子(Nano‐PICsome)の開発. |
| 129. |
Formation of polymer vesicles from microdroplets of polyion complex and examination of their physicochemical properties in microfluidic chamber
This paper presents a direct observation of a non-trivial formation process of individual giant polymer vesicles, i.e., direct formation of vesicles from microdroplets of polyion complex (PIC) and the optimum condition for the formation of the PIC vesicles, providing a basis for an efficient mass-production. In addition, encapsulation of guest polymers and their retention within the PIC vesicle are demonstrated as well as modification of property of the PIC membrane by crosslinking of the constituent polymers. All experiments were achieved in a microfluidic chamber which can control solution conditions with keeping targeted objects under the view field of the optical microscope.. |
| 130. |
ブロック共重合体からなる自己組織化中空粒子PICsomeの開発とin vivo応用. |
| 131. |
架橋率の異なるポリイオンコンプレックス型中空粒子(Nano‐PICsome)の基礎物性評価と体内動態評価. |
| 132. |
Yasutaka Anraku, Akihiro Kishimura, Atsushi Kobayashi, Makoto Oba, Kazunori Kataoka, Size-controlled long-circulating PICsome as a ruler to measure critical cut-off disposition size into normal and tumor tissues, Chemical Communications, 10.1039/c1cc11465d, 47, 21, 6054-6056, 2011.06, Selective disposition of nanocarriers into target tissue is an essential issue in drug delivery. Critical size of nanocarriers (∼150 nm) discriminating the permeability into normal and tumor tissues was determined by the use of size-tunable, polyion complex hollow vesicles (PICsome) as a ruler.. |
| 133. |
Enjoy Conferences!. |
| 134. |
Kosuke Okeyoshi, Daisuke Suzuki, Akihiro Kishimura, Ryo Yoshida, Photoinduced Hydrogen-Generating Nanogel Systems, SMALL, 10.1002/smll.201000872, 7, 3, 311-315, 2011.02. |
| 135. |
H. Oana, M. Morinaga, Akihiro Kishimura, M. Gel, K. Kataoka, M. Washizu, Formation of polymer vesicles from microdroplets of polyion complex and examination of their physicochemical properties in microfluidic chamber, 15th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2011, MicroTAS 2011
15th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2011, MicroTAS 2011, 3, 1588-1590, 2011, This paper presents a direct observation of a non-trivial formation process of individual giant polymer vesicles, i.e., direct formation of vesicles from microdroplets of polyion complex (PIC) and the optimum condition for the formation of the PIC vesicles, providing a basis for an efficient mass-production. In addition, encapsulation of guest polymers and their retention within the PIC vesicle are demonstrated as well as modification of property of the PIC membrane by crosslinking of the constituent polymers. All experiments were achieved in a microfluidic chamber which can control solution conditions with keeping targeted objects under the view field of the optical microscope.. |
| 136. |
Sachiko Kaida, Horacio Cabral, Michiaki Kumagai, Akihiro Kishimura, Yasuko Terada, Masaki Sekino, Ichio Aoki, Nobuhiro Nishiyama, Toru Tani, Kazunori Kataoka, Visible Drug Delivery by Supramolecular Nanocarriers Directing to Single-Platformed Diagnosis and Therapy of Pancreatic Tumor Model, CANCER RESEARCH, 10.1158/0008-5472.CAN-10-0303, 70, 18, 7031-7041, 2010.09, Nanoparticle therapeutics are promising platforms for cancer therapy. However, it remains a formidable challenge to assess their distribution and clinical efficacy for therapeutic applications. Here, by using multifunctional polymeric micellar nanocarriers incorporating clinically approved gadolinium (Gd)-based magnetic resonance imaging contrast agents and platinum (Pt) anticancer drugs through reversible metal chelation of Pt, simultaneous imaging and therapy of an orthotopic animal model of intractable human pancreatic tumor was successfully performed without any serious toxicity. The strong tumor contrast enhancement achieved by the micelles correlated with the 24 times increase of r(1) of the Gd chelates, the highest for the formulations using clinically approved Gd chelates reported to date. From the micro-synchrotron radiation X-ray fluorescence spectrometry scanning of the lesions, we confirmed that both the Gd chelates and Pt drugs delivered by the micelles selectively colocalized in the tumor interior. Our study provides new insights for the design of theranostic micelles with high contrast enhancement and site-specific clinical potential. Cancer Res; 70(18); 7031-41. (C)2010 AACR.. |
| 137. |
Structural control of nano-/micro-capsules through the simple preparation methods based on macromolecular self-assemblies and their application to biomaterials. |
| 138. |
Yasutaka Anraku, Akihiro Kishimura, Makoto Oba, Yuichi Yamasaki, Kazunori Kataoka, Spontaneous Formation of Nanosized Unilamellar Polyion Complex Vesicles with Tunable Size and Properties, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 10.1021/ja908350e, 132, 5, 1631-1636, 2010.02, Fabrication of monodispersed, submicrometer-sized vesicles (nanosomes) that form through self-assembly possessing a thin and permeable membrane remains a significant challenge. Conventional fabrication of nanosomes through self-assembly of amphiphilic molecules often requires cumbersome processes using organic solvents combined with physical procedures (e.g., sonication, thermal treatment, and membrane filtration) to obtain unilamellar structures with a controlled size distribution. Herein, we report the first example of spontaneously formed submicrometer-sized unilamellar polyion complex vesicles (Nano-PICsomes) via self-assembly of a pair of oppositely charged PEG block aniomer and homocatiomer in an aqueous medium. Detailed dynamic light scattering and transmission electron microscopic analysis revealed that vesicle sizes can be controlled in the range of 100-400 nm with a narrow size distribution, simply by changing the total polymer concentration. Also, each Nano-PICsome was composed of a uniform single PIC membrane, the thickness of which is around 10-15 nm, regardless of its size. Fluorescence correlation spectroscopy measurement verified that Nano-PICsomes were able to encapsulate water-soluble fluorescent macromolecules in the inner water phase and release them slowly into the exterior. Moreover, cross-linking of the vesicle membrane allows tuning of permeability, enhancement in stability under physiological conditions, and preservation of size and structure even after freeze-drying and centrifugation treatment. Finally, Nano-PICsomes showed a long circulation time in the bloodstream of mice. Precise control of the particle size and structure of hollow capsules through simple aqueous self-assembly and easy modification of their properties by cross-linking is quite novel and fascinating in terms of ecological, low-cost, and low-energy fabrication processes as well as the potential utility in the biomedical arena.. |
| 139. |
Spontaneous formation of giant unilamellar vesicles from microdroplets of a polyion complex by focused infrared laser irradiation
Spontaneous formation of a giant polymer vesicle from a single micrometer-sized droplet of polyion complex (PIC) of diblock copolymers and its derivative by thermal perturbation, which is achieved by irradiation with a focused infrared laser is presented. The thermal perturbation induces a microphase separation inside of the PIC droplet and the generated water rich phase in the PIC droplet becomes a content of the vesicle and the PIC is deformed into a self-assembled membrane of the vesicle. The size of the giant unilamellar vesicles formed is determined on the basis of the initial size of the PIC droplets. ©2009 IEEE.. |
| 140. |
Hidehiro Oana, Akihiro Kishimura, Kei Yonehara, Yuichi Yamasaki, Masao Washizu, Kazunori Kataoka, Spontaneous formation of giant unilamellar vesicles from microdroplets of a polyion complex by thermally induced phase separation, Angewandte Chemie - International Edition, 10.1002/anie.200900721, 48, 25, 4613-4616, 2009.06, Water pump: Polyion complex (PIC) vesicles are spontaneously formed from PIC microdroplets, which are formed by mixing cationic and anionic polymers (see picture). The formation process can be reversibly controlled by local heating with a focused infrared laser that triggers microphase separation and subsequent water influx. The size of the resulting giant unilamellar vesicles is determined by the initial size of the PIC droplets.. |
| 141. |
CABRAL Horacio, ANRAKU Yasutaka, YAMAMOTO Megumi, KISHIMURA Akihiro, OSADA Kensuke, NISHIYAMA Nobuhiro, KATAOKA Kazunori, Polymeric vesicles assembled by metal complexation as versatile theranostic nanodevices, 高分子学会予稿集(CD-ROM), 58, 1 Disk1, 2L03, 2009.05. |
| 142. |
Wen-Fei Dong, Akihiro Kishimura, Yasutaka Anraku, Sayan Chuanoi, Kazunori Kataoka, Monodispersed Polymeric Nanocapsules: Spontaneous Evolution and Morphology Transition from Reducible Hetero-PEG PICmicelles by Controlled Degradation, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 10.1021/ja808419b, 131, 11, 3804-+, 2009.03, In this communication, a novel "self-templating" strategy was used to prepare uniform and biocompatible nanocapsules by the addition of a reduction agent (i.e., DTT) into a solution of highly monodispersed PICmicelles bearing a heterodetachable PEG Corona. PEG chains were released from PICmicelle shells following disulfide reduction which leads a spontaneous and drastic morphology evolution from micelles to vesicles induced by the decrease of the PEG weight fraction. Formation of uniform nanocapsules with controllable capsule size was achieved by careful control of the micelle composition and molecular weight of homo-P[Asp(DET)].. |
| 143. |
Hidehiro Oana, Akihiro Kishimura, Yuichi Yamasaki, Masao Washizu, Kazunori Kataoka, Spontaneous formation of giant unilamellar vesicles from microdroplets of a polyion complex by focused infrared laser irradiation, 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science
20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science, 10.1109/MHS.2009.5352051, 155-160, 2009, Spontaneous formation of a giant polymer vesicle from a single micrometer-sized droplet of polyion complex (PIC) of diblock copolymers and its derivative by thermal perturbation, which is achieved by irradiation with a focused infrared laser is presented. The thermal perturbation induces a microphase separation inside of the PIC droplet and the generated water rich phase in the PIC droplet becomes a content of the vesicle and the PIC is deformed into a self-assembled membrane of the vesicle. The size of the giant unilamellar vesicles formed is determined on the basis of the initial size of the PIC droplets.. |
| 144. |
Akihiro Kishimura, Sittipong Liamsuwan, Hiroyuki Matsuda, Wen Fei Dong, Kensuke Osada, Yuichi Yamasaki, Kazunori Kataoka, PH-dependent permeability change and reversible structural transition of PEGylated polyion complex vesicles (PICsomes) in aqueous media, Soft Matter, 10.1039/b815884c, 5, 3, 529-532, 2008.11, The acidic pH-sensitivity of polyion complex vesicles (PICsomes) was investigated, using dynamic light scattering (DLS) and confocal laser scanning microscopy (CLSM). PICsomes showed pH-dependent and reversible structural transition, and also underwent a change in permeability by sensing acidic pH. Increased membrane permeability at the pH corresponding to cellular endosomes may be useful for future applications of PICsomes as a delivery vehicle of biologically active compounds to intracellular compartment.. |
| 145. |
Spontaneous formation of polymer vesicles from microdroplets of polyion complex via microphase separation
This paper presents a new method for the formation of giant polymer vesicles, which utilizes a microphase separation occurring in microdroplets of polyion complex (PIC), and continuous production of the giant polymer vesicles is demonstrated using a microfluidic channel. © 2008 CBMS.. |
| 146. |
H. Oana, K. Yonehara, Akihiro Kishimura, Y. Yamasaki, K. Kataoka, M. Washizu, Spontaneous formation of polymer vesicles from microdroplets of polyion complex via microphase separation, 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008
12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference, 631-633, 2008, This paper presents a new method for the formation of giant polymer vesicles, which utilizes a microphase separation occurring in microdroplets of polyion complex (PIC), and continuous production of the giant polymer vesicles is demonstrated using a microfluidic channel.. |
| 147. |
Yuan Li, Woo-Dong Jang, Nobuhiro Nishiyama, Akihiro Kishimura, Satoko Kawauchi, Yuji Morimoto, Sayaka Miake, Takashi Yamashita, Makoto Kikuchi, Takuzo Aida, Kazunori Kataoka, Dendrimer generation effects on photodynamic efficacy of dendrimer porphyrins and dendrimer-loaded supramolecular nanocarriers, CHEMISTRY OF MATERIALS, 10.1021/cm071451m, 19, 23, 5557-5562, 2007.11, A series of poly(benzyl ether) dendrimer porphyrins (DPs) (Gn = n-generation dendrimer, n = 1-3) was examined as potential photosensitizers for photodynamic therapy (PDT). Polyion complexes (PICs) between the DPs and poly(ethylene glycol)-block-poly(L-lysine) (PEG-beta-PLL) were formed via an electrostatic interaction between the positively charged poly(L-lysine) (PLL) segment and negatively charged periphery of the DPs. Dynamic light scattering (DLS) measurements and transmission electron microscopy. (TEM) showed that G3 formed a core-shell-type nanocarrier micelle, whereas G1 and G2 formed irregular-shaped nanoparticles with a relatively high polydispersity. The photophysical properties of the DP-loaded PIC nanocarriers strongly depend on the generation of the DPs. In the case of GI and G2, their fluorescence lifetime and oxygen consumption ability were significantly reduced by the formation of the PIC nanocarriers, whereas the G34oaded PIC nanocarrier exhibited almost comparable fluorescence lifetimes and oxygen consumption abilities to the free G3. The incorporation of DPs into PIC nanocarriers resulted in an appreciable increase in the cellular uptake, yet inversely correlated with the generation. Alternatively, the photocytotoxicity of the DPs within the nanocarriers increased with an increase in the generation despite a decrease in the cellular uptake. By correlating the effects of the uptake amount with the photocytotoxicity, the PIC nanocarriers showed remarkable enhancement of the PDT efficacy dependent on the generation of DPs.. |
| 148. |
Akihiro Kishimura, Aya Koide, Kensuke Osada, Yuichi Yamasaki, Kazunori Kataoka, Encapsulation of myoglobin in PEGylated polyion complex vesicles made from a pair of oppositely charged block ionomers
A physiologically available oxygen carrier, Angewandte Chemie - International Edition, 10.1002/anie.200701776, 46, 32, 6085-6088, 2007.08, (Figure Presented) Take your PIC: Biologically active polyion complex vesicles (PICsomes) with encapsulated myoglobin (Mb) can be prepared by the self-assembly of a pair of oppositely charged block ionomers with polyethylene glycol (PEG) segments (see picture; metMb: metmyoglobin). The loaded Mb maintains reversible oxygenation even in the presence of trypsin.. |
| 149. |
Thermally Rewritable Phosphorescent Paper. |
| 150. |
A Koide, A Kishimura, K Osada, WD Jang, Y Yamasaki, K Kataoka, Semipermeable polymer vesicle (PICsome) self-assembled in aqueous medium from a pair of oppositely charged block copolymers: Physiologically stable micro-/nanocontainers of water-soluble macromolecules, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 10.1021/ja057993r, 128, 18, 5988-5989, 2006.05. |
| 151. |
Tomovuki Kishi, Joon Sik Park, Akihiro Kishimura, Kazunori Kataoka, Synthesis of poly(2-oxazoline)s block copolymers with different functional groups at both terminal and the evaluation of their thermo-sensitive behaviors, 55th Society of Polymer Science Japan Symposium on Macromolecules
55th SPSJ Symposium on Macromolecules, 55, 5048-5049, 2006, In this study, using thermo-sensitive poly (2-isopropyl-2-oxazoline)s (PiPrOx), we prepared a series of block copolymers (PEtOx-e-PiPrOx). Two different functional groups at both terminal ends were introduced to the block copolymers. We investigated the thermo-sensitive behavior of block copolymers and the effect of their compositions and the end functional groups.. |
| 152. |
Tomovuki Kishi, Joon Sik Park, Akihiro Kishimura, Yuichi Yamasaki, Kazunori Kataoka, Synthesis of the thermosensitive poly (oxazolines) and formation of the aggregates, 55th SPSJ Annual Meeting
Polymer Preprints, Japan - 55th SPSJ Annual Meeting, 55, 567, 2006, In this study, we synthesized poly (2-ethyl-2-oxazolines) (PEtOx). And using thermosensitive polymer (poly (2-isopropyl-2-oxazolines) (PiPrOx)), we prepared a series of block copolymers composed of PEtOx and PiPrOx. These copolymers form the aggregates by the hydrophobic interaction near the LCST We investigated the thermosensitive behavior of diblock copolymer (PEtOx (44)-PiPrOx (44)). The aggregates formed at the temperature near the LCST were observed by TEM. We confirmed the aggregates formation responding the temperature.. |
| 153. |
Akihiro Kishimura, Takashi Yamashita, Kentaro Yamaguchi, Takuzo Aida, “Rewritable phosphorescent paper” by the control of competing kinetic and thermodynamic self–assembling events., Nature Materials, 10.1038/nmat1401, 4, 7, 546-549, 2005.07. |
| 154. |
Akihiro Kishimura, Takashi Yamashita, Takuzo Aida, Phosphorescent Organogels via 'Metallophilic' Interactions for Reversible RGB–Color Switching, Journal of the American Chemical Society, 10.1021/ja0441007, 127, 1, 179-183, 2005.01. |
| 155. |
Akihiro Kishimura, Aya Koide, Joon Sik Park, Kensuke Osada, Yuichi Yamasaki, Kazunori Kataoka, The design of "PICsome" in aqueous medium based on oppositely charged block copolymers, 54th SPSJ Symposium on Macromolecules
54th SPSJ Symposium on Macromolecules - Polymer Preprints, Japan, 54, 3529, 2005, A novel type of polymersome formed through a self-assembly of polyion complex of block copolymers was firstly designed as "PICsome". The PICsome can be simply prepared in an aqueous solution via electrostatic interaction between a pair of the oppositely charged block copolymers. A series of copolymers was newly synthesized, and the polymer assemblies were prepared and investigated.. |
| 156. |
Molecular Design of Nano-scale Catalysts Using Dendrimer Scaffolds. |
| 157. |
M Enomoto, A Kishimura, T Aida, Coordination metallacycles of an achiral dendron self-assemble via metal-metal interaction to form luminescent superhelical fibers, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 10.1021/ja010426t, 123, 23, 5608-5609, 2001.06. |
