| 1. |
Fahmida Habib Nabila, Rashedul Islam, Islam Md Shimul, Muhammad Moniruzzaman, Rie Wakabayashi, Noriho Kamiya, Masahiro Goto, Ionic liquid-mediated ethosome for transdermal delivery of insulin., Chemical communications (Cambridge, England), 10.1039/d3cc06130b, 2024.03, Herein, we report ethosome (ET) formulations composed of a safe amount of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC)-based ionic liquid with various concentrations of ethanol as a carrier for the transdermal delivery of a high molecular weight drug, insulin. The Insulin-loaded ET vesicles exhibited long-term stability compared to conventional DMPC ETs, showing significantly higher drug encapsulation efficiency and increased skin permeation ability.. |
| 2. |
Ryutaro Ariyoshi, Takashi Matsuzaki, Ryo Sato, Kosuke Minamihata, Kounosuke Hayashi, Taisei Koga, Kensei Orita, Riko Nishioka, Rie Wakabayashi, Masahiro Goto, Noriho Kamiya, Engineering the Propeptide of Microbial Transglutaminase Zymogen: Enabling Substrate-Dependent Activation for Bioconjugation Applications., Bioconjugate chemistry, 10.1021/acs.bioconjchem.3c00544, 2024.02, Microbial transglutaminase (MTG) from Streptomyces mobaraensis is a powerful biocatalytic glue for site-specific cross-linking of a range of biomolecules and synthetic molecules that have an MTG-reactive moiety. The preparation of active recombinant MTG requires post-translational proteolytic digestion of a propeptide that functions as an intramolecular chaperone to assist the correct folding of the MTG zymogen (MTGz) in the biosynthesis. Herein, we report engineered active zymogen of MTG (EzMTG) that is expressed in soluble form in the host Escherichia coli cytosol and exhibits cross-linking activity without limited proteolysis of the propeptide. We found that the saturation mutagenesis of residues K10 or Y12 in the propeptide domain generated several active MTGz mutants. In particular, the K10D/Y12G mutant exhibited catalytic activity comparable to that of mature MTG. However, the expression level was low, possibly because of decreased chaperone activity and/or the promiscuous substrate specificity of MTG, which is potentially harmful to the host cells. The K10R/Y12A mutant exhibited specific substrate-dependent reactivity toward peptidyl substrates. Quantitative analysis of the binding affinity of the mutated propeptides to the active site of MTG suggested an inverse relationship between the binding affinity and the catalytic activity of EzMTG. Our proof-of-concept study provides insights into the design of a new biocatalyst using the MTGz as a scaffold and a potential route to high-throughput screening of EzMTG mutants for bioconjugation applications.. |
| 3. |
Diah Anggraini Wulandari, Kyosuke Tsuru, Kosuke Minamihata, Rie Wakabayashi, Masahiro Goto, Noriho Kamiya, A Functional Hydrogel Bead-Based High-Throughput Screening System for Mammalian Cells with Enhanced Secretion of Therapeutic Antibodies., ACS biomaterials science & engineering, 10.1021/acsbiomaterials.3c01386, 10, 1, 628-636, 2024.01, Droplet-based high-throughput screening systems are an emerging technology that provides a quick test to screen millions of cells with distinctive characteristics. Biopharmaceuticals, specifically therapeutic proteins, are produced by culturing cells that secrete heterologous recombinant proteins with different populations and expression levels; therefore, a technology to discriminate cells that produce more target proteins is needed. Here, we present a droplet-based microfluidic strategy for encapsulating, screening, and selecting target cells with redox-responsive hydrogel beads (HBs). As a proof-of-concept study, we demonstrate the enrichment of hybridoma cells with enhanced capability of antibody secretion using horseradish peroxidase (HRP)-catalyzed hydrogelation of tetra-thiolate poly(ethylene glycol); hybridoma cells were encapsulated in disulfide-bonded HBs. Recombinant protein G or protein M with a C-terminal cysteine residue was installed in the HBs via disulfide bonding to capture antibodies secreted from the cells. HBs were fluorescently stained by adding the protein L-HRP conjugate using a tyramide signal amplification system. HBs were then separated by fluorescence-activated droplet sorting and degraded by reducing the disulfide bonds to recover the target cells. Finally, we succeeded in the selection of hybridoma cells with enhanced antibody secretion, indicating the potential of this system in the therapeutic protein production.. |
| 4. |
Chowdhury Md Raihan, Rahman Md. Moshikur, Rie Wakabayashi, Muhammad Moniruzzaman, Masahiro Goto, Biocompatible ionic liquids assisted transdermal co-delivery of antigenic protein and adjuvant for cancer immunotherapy, International Journal of Pharmaceutics, 10.1016/j.ijpharm.2021.120582, 601, 2021.05. |
| 5. |
Md. Korban Ali, Rahman Md. Moshikur, Rie Wakabayashi, Muhammad Moniruzzaman, Masahiro Goto, Biocompatible Ionic Liquid-Mediated Micelles for Enhanced Transdermal Delivery of Paclitaxel, ACS Applied Materials Interfaces, 10.1021/acsami.1c03111, 13, 17, 19745-19755, 2021.05. |
| 6. |
Mari Takahara, Shinichi Mochizuki, Rie Wakabayashi, Kosuke Minamihata, Masahiro Goto, Kazuo Sakurai, Noriho Kamiya, Extending the Half-Life of a Protein in Vivo by Enzymatic Labeling with Amphiphilic Lipopeptides, Bioconjugate Chemistry, 10.1021/acs.bioconjchem.1c00027, 32, 4, 655-660, 2021.03. |
| 7. |
Rie Wakabayashi, Ayato Higuchi, Hiroki Obayashi, Masahiro Goto, Noriho Kamiya, pH-Responsive Self-Assembly of Designer Aromatic Peptide Amphiphiles and Enzymatic Post-Modification of Assembled Structures, International Journal of Molecular Sciences, 10.3390/ijms22073459, 22, 7, 2021.03. |
| 8. |
Yuya Hirakawa, Hiroshi Ueda, Yusuke Takata, Kosuke Minamihata, Rie Wakabayashi, Noriho Kamiya, Masahiro Goto, Co-amorphous formation of piroxicam-citric acid to generate supersaturation and improve skin permeation., European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences, 10.1016/j.ejps.2020.105667, 158, 105667-105667, 2021.03, The objective of this study was to prepare a co-amorphous formulation of piroxicam (PIR), a non-steroidal anti-inflammatory drug, and citric acid (CA), and evaluate its skin permeation ability. A spray-drying method was employed to prepare the co-amorphous formulation and its physical properties were characterized. X-ray powder diffraction and thermal analysis confirmed a homogeneous amorphous state, and the infrared spectra revealed intermolecular interactions between PIR and CA, suggesting formation of a co-amorphous formulation of PIR and CA. The PIR-CA co-amorphous formulation exhibited no crystallization for 60 days at 4/25/40°C with silica gel. The PIR-CA co-amorphous formulation increased the solubility of PIR in polyethylene glycol 400 compared with that of the pure drug, and physical mixture (PM) of PIR and CA, confirming a supersaturated state in the formulation. The PIR-CA co-amorphous formulation demonstrated higher skin permeation than PIR alone or PM of PIR and CA, and the flux value was consistent with the degree of saturation. Thus, the increase in the skin permeation of PIR from the PIR-CA co-amorphous formulation directly depended on the increased thermodynamic activity by supersaturation in the absence of interactions between the drug and co-former in the vehicle.. |
| 9. |
Wahyu Ramadhan, Genki Kagawa, Kousuke Moriyama, Rie Wakabayashi, Kosuke Minamihata, Masahiro Goto, Noriho Kamiya, Construction of higher-order cellular microstructures by a self-wrapping co-culture strategy using a redox-responsive hydrogel, Scientific reports, 10.1038/s41598-020-63362-4, 10, 1, 2020.12, In this report, a strategy for constructing three-dimensional (3D) cellular architectures comprising viable cells is presented. The strategy uses a redox-responsive hydrogel that degrades under mild reductive conditions, and a confluent monolayer of cells (i.e., cell sheet) cultured on the hydrogel surface peels off and self-folds to wrap other cells. As a proof-of-concept, the self-folding of fibroblast cell sheet was triggered by immersion in aqueous cysteine, and this folding process was controlled by the cysteine concentration. Such folding enabled the wrapping of human hepatocellular carcinoma (HepG2) spheroids, human umbilical vein endothelial cells and collagen beads, and this process improved cell viability, the secretion of metabolites and the proliferation rate of the HepG2 cells when compared with a two-dimensional culture under the same conditions. A key concept of this study is the ability to interact with other neighbouring cells, providing a new, simple and fast method to generate higher-order cellular aggregates wherein different types of cellular components are added. We designated the method of using a cell sheet to wrap another cellular aggregate the ‘cellular Furoshiki’. The simple self-wrapping Furoshiki technique provides an alternative approach to co-culture cells by microplate-based systems, especially for constructing heterogeneous 3D cellular microstructures.. |
| 10. |
Rahman Md Moshikur, Md. Korban Ali, Rie Wakabayashi, Muhammad Moniruzzaman, Masahiro Goto, Formation and potential application of micelles composed of biocompatible N-lauroyl-amino acid ionic liquids surfactant, Journal of Molecular Liquids, 10.1016/j.molliq.2020.114424, 320, 114424, 2020.12. |
| 11. |
Wahyu Ramadhan, Yuki Ohama, Kosuke Minamihata, Kousuke Moriyama, Rie Wakabayashi, Masahiro Goto, Noriho Kamiya, Redox-responsive functionalized hydrogel marble for the generation of cellular spheroids, Journal of Bioscience and Bioengineering, 10.1016/j.jbiosc.2020.05.010, 130, 4, 416-423, 2020.10, Liquid marbles (LMs) have recently shown a great promise as microbioreactors to construct self-supported aqueous compartments for chemical and biological reactions. However, the evaporation of the inner aqueous liquid core has limited their application, especially in studying cellular functions. Hydrogels are promising scaffolds that provide a spatial environment suitable for three-dimensional cell culture. Here, we describe the fabrication of redox-responsive hydrogel marbles (HMs) as a three-dimensional cell culture platform. The HMs are prepared by introducing an aqueous mixture of a tetra-thiolated polyethylene glycol (PEG) derivative, thiolated gelatin (Gela-SH), horseradish peroxidase, a small phenolic compound, and human hepatocellular carcinoma cells (HepG2) to the inner aqueous phase of LMs. Eventually, HepG2 cells are encapsulated in the HMs then immersed in culture media, where they proliferate and form cellular spheroids. Experimental results show that the Gela-SH concentration strongly influences the physicochemical and microstructure properties of the HMs. After 6 days in culture, the spheroids were recovered from the HMs by degrading the scaffold, and examination showed that they had reached up to about 180 μm in diameter depending on the Gela-SH concentration, compared with 60 μm in conventional HMs without Gela-SH. After long-term culture (over 12 days), the liver-specific functions (secretion of albumin and urea) and DNA contents of the spheroids cultured in the HMs were elevated compared with those cultured in LMs. These results suggest that the developed HMs can be useful in designing a variety of microbioreactors for tissue engineering applications.. |
| 12. |
Shihab Uddin, Md Raihan Chowdhury, Rie Wakabayashi, Noriho Kamiya, Muhammad Moniruzzaman, Masahiro Goto, Lipid based biocompatible ionic liquids: synthesis, characterization and biocompatibility evaluation, Chemical Communications, 10.1039/D0CC04491A, 56, 13756-13759, 2020.10. |
| 13. |
Kosuke Minamihata, Yusei Hamada, Genki Kagawa, Wahyu Ramadhan, Ayato Higuchi, Kousuke Moriyama, Rie Wakabayashi, Masahiro Goto, Noriho Kamiya, Dual-functionalizable streptavidin-SpyCatcher fused protein-polymer hydrogels as scaffolds for cell culture, ACS Applied Bio Materials, 10.1021/acsabm.0c00940, 3, 7734-7742, 2020.10. |
| 14. |
Yoshiro Tahara, Kaho Morita, Rie Wakabayashi, Noriho Kamiya, Masahiro Goto, Biocompatible Ionic Liquid Enhances Transdermal Antigen Peptide Delivery and Preventive Vaccination Effect, Molecular pharmaceutics, 10.1021/acs.molpharmaceut.0c00598, 2020.09, Ionic liquids (ILs) attract significant attention as novel solvents for drug delivery systems because of their ability to solubilize poorly soluble drugs and tune the physiological properties of active pharmaceutical ingredients. For the next generation of IL-based drug delivery systems, biocompatibility is a high priority. In the current study, choline-fatty acids ([Cho][FA]) were used as a biocompatible IL to mediate the dissolution of a water-soluble antigen peptide in an oil-based skin penetration enhancer. Among the candidate fatty acids (C8, C10, C12, C14, C16, C18:0, and C18:1), C18:1 was selected because of its low cytotoxicity and mediation of skin permeability for an antigen peptide. Using IL[Cho][C18:1] and an oil-based penetration enhancer, the flux of transdermal delivery of the peptide increased 28-fold compared with delivery using an aqueous vehicle. Furthermore, the IL-mediated transcutaneous vaccination succeeded in suppressing tumor growth in vivo compared to injection. The skin irritation produced by this formulation was tested using an in vitro 3D constructed skin tissue model and an in vivo histological study, which concluded that the formulation did not cause skin irritation. The results suggest that biocompatible IL-mediated dissolution in an oil-based skin penetration enhancer is a promising strategy for transdermal drug delivery.. |
| 15. |
Shuto Kozaka, Ayaka Kashima, Rie Wakabayashi, Takahiro Nakata, Taro Ueda, Masahiro Goto, Effective transcutaneous delivery of hyaluronic acid using an easy-to-prepare reverse micelle formulation, Cosmetics, 10.3390/COSMETICS7030052, 7, 3, 2020.09, The skin loses its moisture with advancing age, causing cosmetic issues such as wrinkles. In addition, the loss of moisture leads to hypersensitivity to external stimuli such as UV light. Transcutaneous supplementation with hyaluronic acid (HA) is an effective and safe method of recovering the moisturizing function and elasticity of the skin. However, the transcutaneous delivery of HA remains challenging owing to the barrier function of the stratum corneum (SC) layer. To penetrate the SC barrier, we used a reverse micelle formulation that does not require high energy consumption processes for preparation. We aimed to enhance the skin permeability of HA by incorporating glyceryl monooleate-a skin permeation enhancer-into the formulation. A fluorescently-labeled HA-loaded reverse micelle formulation showed significantly enhanced permeation across Yucatan micro pig skin. Fourier transform infra-red spectroscopy of the surface of the skin treated with the reverse micelle formulation showed blue shifts of the CH2 symmetric/asymmetric stretching peaks, indicating a reduction in the barrier function of the SC. Further study revealed that HA was released from the reverse micelles at the hydrophobic/hydrophilic interface between the SC and the living epidermis. The results demonstrated that our reverse micellar system is an easy-to-prepare formulation for the effective transcutaneous delivery of HA.. |
| 16. |
Rahman Md Moshikur, Md Raihan Chowdhury, Hiroki Fujisawa, Rie Wakabayashi, Muhammad Moniruzzaman, Masahiro Goto, Design and characterization of fatty acid-based amino acid ester as a new “green” hydrophobic ionic liquid for drug delivery, ACS Sustainable Chemistry & Engineering, 10.1021/acssuschemeng.0c03419, 8, 13660-13671, 2020.08. |
| 17. |
R. Sato, K. Minamihata, R. Wakabayashi, M. Goto, N. Kamiya, PolyTag
A peptide tag that affords scaffold-less covalent protein assembly catalyzed by microbial transglutaminase: Design of PolyTag for scaffold-less covalent protein assembly, Analytical Biochemistry, 10.1016/j.ab.2020.113700, 600, 2020.07, Assembling proteins in close vicinity to each other provides an opportunity to gain unique function because collaborative and even synergistic functionalities can be expected in an assembled form. There have been a variety of strategies to synthesize functional protein assemblies but site-specific covalent assembly of monomeric protein units without impairing their intrinsic function remains challenging. Herein we report a powerful strategy to design protein assemblies by using microbial transglutaminase (MTG). A serendipitous discovery of self-crosslinking of enhanced green fluorescent protein (EGFP) fused with StrepTag I at the C-terminus revealed that EGFP was assembled through the crosslinking of the Lys (K) residue in the C-terminus of EGFP and the Gln (Q) residue in StrepTag I (AWRHPQFGG). Site-directed mutagenesis of the residues next to the K and Q yielded EGFP assemblies with higher molecular weights. An optimized peptide tag comprised of both K and Q residues (HKRWRHYQRGG) enabled the assembly of different types of proteins of interest (POI) when it was fused to either the N- or C-terminus. The peptide tag that enabled the self-polymerization of the functional POI without a scaffold was designated as a ‘PolyTag’.. |
| 18. |
Md Rafiqul Islam, Md Raihan Chowdhury, Rie Wakabayashi, Yoshiro Tahara, Noriho Kamiya, Muhammad Moniruzzaman, Masahiro Goto, Choline and amino acid based biocompatible ionic liquid mediated transdermal delivery of the sparingly soluble drug acyclovir, International Journal of Pharmaceutics, 10.1016/j.ijpharm.2020.119335, 582, 2020.05, Transdermal delivery of drugs is more challenging for drugs that are insoluble or sparingly soluble in water and most organic solvents. To overcome this problem, ionic liquid (IL)-mediated ternary systems have been suggested as potential drug carriers. Here, we report potent ternary (IL–EtOH–IPM) systems consisting of biocompatible ILs, ethanol (EtOH), and isopropyl myristate (IPM) that can dissolve a significant amount of the sparingly soluble drug acyclovir (ACV). The ternary systems were optically transparent and thermodynamically stable with a wide range of IL pertinence. An in vitro drug permeation study showed that the ILs in the ternary systems dramatically enhanced ACV permeation into and across the skin. Fourier Transform Infrared spectroscopy of the stratum corneum (sc) after treatment with ternary systems showed that the skin barrier function was reduced by disturbance of the regularly ordered arrangement of corneocytes and modification of the surface properties of the sc during permeation. Histological analysis, and skin irritation studies using a reconstructed human epidermis model showed the safety profile of the ternary system, and there were no significant changes in the structures of the sc, epidermis, and dermis. Therefore, ternary systems containing biocompatible ILs are promising for transdermal delivery of insoluble or sparingly soluble drugs.. |
| 19. |
Md Rafiqul Islam, Md Raihan Chowdhury, Rie Wakabayashi, Noriho Kamiya, Muhammad Moniruzzaman, Masahiro Goto, Ionic liquid-in-oil microemulsions prepared with biocompatible choline carboxylic acids for improving the transdermal delivery of a sparingly soluble drug, Pharmaceutics, 10.3390/pharmaceutics12040392, 12, 4, 2020.04, The transdermal delivery of sparingly soluble drugs is challenging due to of the need for a drug carrier. In the past few decades, ionic liquid (IL)-in-oil microemulsions (IL/O MEs) have been developed as potential carriers. By focusing on biocompatibility, we report on an IL/O ME that is designed to enhance the solubility and transdermal delivery of the sparingly soluble drug, acyclovir. The prepared MEs were composed of a hydrophilic IL (choline formate, choline lactate, or choline propionate) as the non-aqueous polar phase and a surface-active IL (choline oleate) as the surfactant in combination with sorbitan laurate in a continuous oil phase. The selected ILs were all biologically active ions. Optimized pseudo ternary phase diagrams indicated the MEs formed thermodynamically stable, spherically shaped, and nano-sized (90%) with the ME compared with Dulbecco’s phosphate-buffered saline, indicates the biocompatibility of the ME. Therefore, we conclude that IL/O ME may be a promising nano-carrier for the transdermal delivery of sparingly soluble drugs.. |
| 20. |
Yuya Hirakawa, Hiroshi Ueda, Rie Wakabayashi, Noriho Kamiya, Masahiro Goto, A novel binary supercooled liquid formulation for transdermal drug delivery, Biological and Pharmaceutical Bulletin, 10.1248/bpb.b19-00642, 43, 3, 393-398, 2020.03, The aim of this study was to prepare binary supercooled liquid (SCL) by intermolecular interaction and apply this formulation to transdermal drug delivery. Ketoprofen (KET) and ethenzamide (ETH) were selected as binary SCL component. Thermal analysis of physical mixtures of KET and ETH showed decreases in melting points and glass transition below room temperature, thereby indicating formation of KET–ETH SCL. Intermolecular interactions between KET and ETH in the SCL were evaluated from Fourier transform (FT)-IR spectra. KET–ETH SCL maintained SCL state at 25°C with silica gel over 31d and at 40°C/89% relative humidity (RH) over 7d. KET SCL and KET–ETH SCL showed similar permeability of KET for hairless mice skin, which was two-fold higher than that of KET aqueous suspension. Our findings suggest that the SCL state could enhance the skin permeation of drugs and the binary SCL formed by intermolecular interaction could also improve the stability of the SCL. The binary SCL system could become a new drug form for transdermal drug delivery.. |
| 21. |
Dani Permana, Kosuke Minamihata, Ryo Sato, Rie Wakabayashi, Masahiro Goto, Noriho Kamiya, Linear Polymerization of Protein by Sterically Controlled Enzymatic Cross-Linking with a Tyrosine-Containing Peptide Loop, ACS Omega, 10.1021/acsomega.9b04163, 5, 10, 5160-5169, 2020.03, The structure of a protein complex needs to be controlled appropriately to maximize its functions. Herein, we report the linear polymerization of bacterial alkaline phosphatase (BAP) through the site-specific cross-linking reaction catalyzed by Trametes sp. laccase (TL). We introduced a peptide loop containing a tyrosine (Y-Loop) to BAP, and the Y-Looped BAP was treated with TL. The Y-Looped BAP formed linear polymers, whereas BAP fused with a C-terminal peptide containing a tyrosine (Y-tag) showed an irregular shape after TL treatment. The sterically confined structure of the Y-Loop could be responsible for the formation of linear BAP polymers. TL-catalyzed copolymerization of Y-Looped BAP and a Y-tagged chimeric antibody-binding protein, pG2pA-Y, resulted in the formation of linear bifunctional protein copolymers that could be employed as protein probes in an enzyme-linked immunosorbent assay (ELISA). Copolymers comprising Y-Looped BAP and pG2pA-Y at a molar ratio of 100:1 exhibited the highest signal in the ELISA with 26- and 20-fold higher than a genetically fused chimeric protein, BAP-pG2pA-Y, and its polymeric form, respectively. This result revealed that the morphology of the copolymers was the most critical feature to improve the functionality of the protein polymers as detection probes, not only for immunoassays but also for other diagnostic applications.. |
| 22. |
Rahman Md Moshikur, Md Raihan Chowdhury, Rie Wakabayashi, Yoshiro Tahara, Noriho Kamiya, Muhammad Moniruzzaman, Masahiro Goto, Ionic liquids with N-methyl-2-pyrrolidonium cation as an enhancer for topical drug delivery
Synthesis, characterization, and skin-penetration evaluation, Journal of Molecular Liquids, 10.1016/j.molliq.2019.112166, 299, 2020.02, The development of non-toxic ionic liquid-based active pharmaceutical ingredients (IL-APIs) for effective topical drug delivery is still challenging. The properties of IL-APIs can be boosted up by selecting potential biocompatible cations. Here, we introduced N-methyl-2-pyrrolidone (NMP) as a potent biocompatible counter ion to prepare ionic liquefied drugs for topical drug delivery. The cytotoxicity of NMP cation was investigated using mammalian cell lines (HepG2, NIH3T3 and L929 cells) and compared with conventional IL-forming cations. The synthesized NMP cation has lower toxicity than that of conventional IL-forming cations. The NMP cation showed at least 3.6, 15.2 and 58.9 times lower toxicity than that of conventional imidazolium, ammonium and phosphonium cations, respectively. The synthesized NMP-based ionic liquid (NMP-IL) was characterized using 1H & 13C NMR, FT-IR, DSC and TGA. NMP-IL showed better physico-thermal stability, enhanced skin penetration, and enriched drug accumulation 2.6 times higher than that of IL [Cho][Ibu] in the target tissue. These results suggested that NMP cation based API-IL can be an effective biocompatible formulation for topical drug delivery by accumulating active drugs in the skin.. |
| 23. |
Shuto Kozaka, Yoshiro Tahara, Rie Wakabayashi, Takahiro Nakata, Taro Ueda, Noriho Kamiya, Masahiro Goto, Transcutaneous Cancer Vaccine Using a Reverse Micellar Antigen Carrier, Molecular pharmaceutics, 10.1021/acs.molpharmaceut.9b01104, 17, 2, 645-655, 2020.02, Skin dendritic cells (DCs) such as Langerhans cells and dermal dendritic cells have a pivotal role in inducing antigen-specific immunity; therefore, transcutaneous cancer vaccines are a promising strategy to prophylactically prevent the onset of a variety of diseases, including cancers. The largest obstacle to delivering antigen to these skin DC subsets is the barrier function of the stratum corneum. Although reverse micellar carriers are commonly used to enhance skin permeability to hydrophilic drugs, the transcutaneous delivery of antigen, proteins, or peptides has not been achieved to date because of the large molecular weight of drugs. To achieve effective antigen delivery to skin DCs, we developed a novel strategy using a surfactant as a skin permeation enhancer in a reverse micellar carrier. In this study, glyceryl monooleate (MO) was chosen as a skin permeation enhancer, and the MO-based reverse micellar carrier enabled the successful delivery of antigen to Langerhans cells and dermal dendritic cells. Moreover, transcutaneous vaccination with the MO-based reverse micellar carrier significantly inhibited tumor growth, indicating that it is a promising vaccine platform against tumors.. |
| 24. |
Qingliang Kong, Momoko Kitaoka, Yoshiro Tahara, Rie Wakabayashi, Noriho Kamiya, Masahiro Goto, Solid-in-oil nanodispersions for intranasal vaccination
Enhancement of mucosal and systemic immune responses, International Journal of Pharmaceutics, 10.1016/j.ijpharm.2019.118777, 572, 2019.12, En masse vaccination is a promising strategy for combatting infectious diseases. Intranasal vaccination is a viable route of mass vaccination, and it could be performed easily via needle-free administration. However, it is not widely used because it tends not to evoke sufficient immunity. The aim of the present study was to improve the performance of intranasal vaccination by extending the amount of time that administered antigens remain in the nasal cavity, and enhancing immune responses via a nanocarrier-based adjuvant. A simple and safe solid-in-oil (S/O) system was investigated as a nanocarrier in intranasal vaccination. S/O nanodispersions are oil-based dispersions of antigens coated with surfactants. Because of the mucoadhesive capacities of surfactant and oil they have high potential to extend the amount of time that administered antigens remain in the nasal cavity, and can induce strong immune responses due to a nanocarrier-based adjuvant effect. In nasal absorption experiments antigens administered intranasally via S/O nanodispersions remained in the nasal cavity longer and induced strong mucosal and systemic immune responses. Histopathology analysis indicated that S/O nanodispersions did not modify the nasal epithelium or cilia, suggesting non-toxicity of the carrier. These results indicate the potential of intranasal vaccination using S/O nanodispersions for future vaccination.. |
| 25. |
Qingliang Kong, Kouki Higasijima, Rie Wakabayashi, Yoshiro Tahara, Momoko Kitaoka, Hiroki Obayashi, Yanting Hou, Noriho Kamiya, Masahiro Goto, Transcutaneous delivery of immunomodulating pollen extract-galactomannan conjugate by solid-in-oil nanodispersions for pollinosis immunotherapy, Pharmaceutics, 10.3390/pharmaceutics11110563, 11, 11, 2019.11, Japanese cedar pollinosis is a type I allergic disease and has already become a major public health problem in Japan. Conventional subcutaneous immunotherapy (SCIT) and sublingual immunotherapy (SLIT) cannot meet patients’ needs owing to the side effects caused by both the use of conventional whole antigen molecules in the pollen extract and the administration routes. To address these issues, a surface-modified antigen and transcutaneous administration route are introduced in this research. First, the pollen extract (PE) was conjugated to galactomannan (PE-GM) to mask immunoglobulin E (IgE)-binding epitopes in the PE to avoid side effects. Second, as a safer alternative to SCIT and SLIT, transcutaneous immunotherapy (TCIT) with a solid-in-oil (S/O) nanodispersion system carrying PE-GM was proposed. Hydrophilic PE-GM was efficiently delivered through mouse skin using S/O nanodispersions, reducing the antibody secretion and modifying the type 1 T helper (Th1)/ type 2 T helper (Th2) balance in the mouse model, thereby demonstrating the potential to alleviate Japanese cedar pollinosis.. |
| 26. |
Md Korban Ali, Rahman Md Moshikur, Rie Wakabayashi, Yoshiro Tahara, Muhammad Moniruzzaman, Noriho Kamiya, Masahiro Goto, Synthesis and characterization of choline–fatty-acid-based ionic liquids
A new biocompatible surfactant, Journal of Colloid And Interface Science, 10.1016/j.jcis.2019.04.095, 551, 72-80, 2019.09. |
| 27. |
Safrina Dyah Hardiningtyas, Seiya Nagao, Emiko Yamamoto, Nana Shirakigawa, Rie Wakabayashi, Masahiro Goto, Hiroyuki Ijima, Noriho Kamiya, A nano-sized gel-in-oil suspension for transcutaneous protein delivery, International Journal of Pharmaceutics, 10.1016/j.ijpharm.2019.118495, 567, 2019.08, We developed a new oil-based delivery system for transdermal protein delivery, a gel-in-oil (G/O) nanosuspension, where gelatin-based hydrogel was coated with hydrophobic surfactants. The high entrapment efficiency of a model protein, phycocyanin (PC), into nano-sized gelatin hydrogel particles was achieved. Spectroscopic evaluation of PC suggested that the G/O nanosuspension could retain the functional form of PC in isopropyl myristate. In vitro skin permeation studies showed that the G/O nanosuspension facilitated the delivery of PC through the stratum corneum of Yucatan micropig skin.. |
| 28. |
Md Raihan Chowdhury, Rahman Md Moshikur, Rie Wakabayashi, Yoshiro Tahara, Noriho Kamiya, Muhammad Moniruzzaman, Masahiro Goto, In vivo biocompatibility, pharmacokinetics, antitumor efficacy, and hypersensitivity evaluation of ionic liquid-mediated paclitaxel formulations, International Journal of Pharmaceutics, 10.1016/j.ijpharm.2019.05.020, 565, 219-226, 2019.06. |
| 29. |
Wahyu Ramadhan, Genki Kagawa, Yusei Hamada, Kousuke Moriyama, Rie Wakabayashi, Kosuke Minamihata, Masahiro Goto, Noriho Kamiya, Enzymatically Prepared Dual Functionalized Hydrogels with Gelatin and Heparin to Facilitate Cellular Attachment and Proliferation, ACS Applied Bio Materials, 10.1021/acsabm.9b00275, 2, 6, 2600-2609, 2019.06, Biologically active artificial scaffolds for cell seeding are developed by mimicking extracellular matrices using synthetic materials. Here, we propose a feasible approach employing biocatalysis to integrate natural components, that is, gelatin and heparin, into a synthetic scaffold, namely a polyethylene glycol (PEG)-based hydrogel. Initiation of horseradish peroxidase-mediated redox reaction enabled both hydrogel formation of tetra-thiolated PEG via disulfide linkage and incorporation of chemically thiolated gelatin (Gela-SH) and heparin (Hepa-SH) into the polymeric network. We found that the compatibility of the type of gelatin with heparin was crucial for the hydrogelation process. Alkaline-treated gelatin exhibited superior performance over acid-treated gelatin to generate dual functionality in the resultant hydrogel originating from the two natural biopolymers. The Gela-SH/Hepa-SH dual functionalized PEG-based hydrogel supported both cellular attachment and binding of basic fibroblast growth factor (bFGF) under cell culture conditions, which increased the proliferation and phenotype transformation of NIH3T3 cells cultured on the hydrogel. Inclusion of bFGF and a commercial growth factor cocktail in hydrogel matrices effectively enhanced cell spreading and confluency of both NIH3T3 cells and HUVECs, respectively, suggesting a potential method to design artificial scaffolds containing active growth factors.. |
| 30. |
Rie Wakabayashi, Hiroki Obayashi, Ryuichiro Hashimoto, Noriho Kamiya, Masahiro Goto, Complementary interaction with peptide amphiphiles guides size-controlled assembly of small molecules for intracellular delivery, Chemical Communications, 10.1039/c9cc02473e, 55, 49, 6997-7000, 2019.06, We introduced complementary interactions between peptide amphiphiles and a small fluorescence dye to develop a programmable multi-component supramolecular assembly, and intracellular delivery of the dye was controlled by the dimensions of the co-assembly, which was manipulated by the peptide design.. |
| 31. |
Rie Wakabayashi, Hidetoshi Kono, Shuto Kozaka, Yoshiro Tahara, Noriho Kamiya, Masahiro Goto, Transcutaneous codelivery of tumor antigen and resiquimod in solid-in-oil nanodispersions promotes antitumor immunity, ACS Biomaterials Science and Engineering, 10.1021/acsbiomaterials.9b00260, 2019.04, Cancer vaccines aim to prevent or inhibit tumor growth by inducing an immune response to tumor-associated antigens (TAAs) encoded by or present in the vaccine. Previous work has demonstrated that effective antitumor immunity can be induced using a codelivery system in which nonspecific immunostimulatory molecules are administered together with TAAs. In this study, we investigated the antitumor effects of a solid-in-oil (S/O) nanodispersion system containing a model TAA, ovalbumin (OVA), and resiquimod (R-848), a small molecular Toll-like receptor 7/8 ligand, which induces an antigen-nonspecific cellular immune response that is crucial for the efficacy of cancer vaccines. R-848 was contained in the outer oil phase of S/O nanodispersion. Analysis of OVA and R-848 permeation in mouse skin after application of an R-848 S/O nanodispersion indicated that R-848 rapidly permeated the skin and preactivated Langerhans cells, resulting in efficient uptake of OVA and migration of antigen-loaded Langerhans cells to the draining lymph nodes. Transcutaneous immunization of mice with an R-848 S/O nanodispersion inhibited the growth of E.G7-OVA tumors and prolonged mouse survival to a greater extent than did immunization with an S/O nanodispersion containing OVA alone. Consistent with this observation, antigen-specific secretion of the Th1 cytokine interferon-γand cytolytic activity were both high in splenocytes isolated from mice immunized with R-848 S/O. Our results thus demonstrate that codelivery of R-848 significantly amplified the antitumor immune response induced by antigen-containing S/O nanodispersions and further suggest that S/O nanodispersions may be effective formulations for codelivery of TAAs and R-848 in transcutaneous cancer vaccines.. |
| 32. |
Rahman Md Moshikur, Md Raihan Chowdhury, Rie Wakabayashi, Yoshiro Tahara, Muhammad Moniruzzaman, Masahiro Goto, Ionic liquids with methotrexate moieties as a potential anticancer prodrug
Synthesis, characterization and solubility evaluation, Journal of Molecular Liquids, 10.1016/j.molliq.2019.01.063, 278, 226-233, 2019.03. |
| 33. |
Muhamad Alif Razi, Rie Wakabayashi, Masahiro Goto, Noriho Kamiya, Self-Assembled Reduced Albumin and Glycol Chitosan Nanoparticles for Paclitaxel Delivery, Langmuir, 10.1021/acs.langmuir.8b02809, 35, 7, 2610-2618, 2019.02. |
| 34. |
Rie Wakabayashi, Ayumi Suehiro, Masahiro Goto, Noriho Kamiya, Designer aromatic peptide amphiphiles for self-assembly and enzymatic display of proteins with morphology control, Chemical Communications, 10.1039/C8CC08163H, 55, 5, 640-643, 2019.01. |
| 35. |
Shuto Kozaka, Rie Wakabayashi, Onofrio Annunziata, Milan Balaz, Masahiro Goto, Noriho Kamiya, Sergei V. Dzyuba, Effect of macromolecular crowding on the conformational behaviour of a porphyrin rotor, Journal of Photochemistry and Photobiology A: Chemistry, 10.1016/j.jphotochem.2018.10.006, 369, 115-118, 2019.01. |
| 36. |
Safrina Dyah Hardiningtyas, Rie Wakabayashi, Ryutaro Ishiyama, Yuki Owada, Masahiro Goto, Noriho Kamiya, Enhanced potential of therapeutic applications of curcumin using solid-in-water nanodispersion technique, JOURNAL OF CHEMICAL ENGINEERING OF JAPAN, 10.1252/jcej.18we060, 52, 1, 138-143, 2019.01. |
| 37. |
Mari Takahara, Rie Wakabayashi, Naoki Fujimoto, Kosuke Minamihata, Masahiro Goto, Noriho Kamiya, Enzymatic Cell-Surface Decoration with Proteins using Amphiphilic Lipid-Fused Peptide Substrates, Chemistry - A European Journal, 10.1002/chem.201900370, 2019.01. |
| 38. |
Md Raihan Chowdhury, Rahman Md Moshikur, Rie Wakabayashi, Yoshiro Tahara, Noriho Kamiya, Muhammad Moniruzzaman, Masahiro Goto, Development of a novel ionic liquid-curcumin complex to enhance its solubility, stability, and activity, Chemical Communications, 10.1039/c9cc02812a, 55, 54, 7737-7740, 2019.01, We report a one-step emulsification and rapid freeze-drying process to develop a curcumin-ionic liquid (CCM-IL) complex that could be readily dispersed in water with a significantly enhanced solubility of ∼8 mg mL-1 and half-life (t1/2) of ∼260 min compared with free CCM (solubility ∼30 nM and t1/2 ∼ 20 min). This process using an IL consisting of a long chain carbon backbone as a surfactant, may provide an alternative way of enhancing the solubility of poorly water-soluble drugs.. |
| 39. |
Noriho Kamiya, Yuki Ohama, Kosuke Minamihata, Rie Wakabayashi, Masahiro Goto, Liquid Marbles as an Easy-to-Handle Compartment for Cell-Free Synthesis and In Situ Immobilization of Recombinant Proteins, Biotechnology Journal, 10.1002/biot.201800085, 13, 12, 2018.12. |
| 40. |
Mari Takahara, Rie Wakabayashi, Kosuke Minamihata, Masahiro Goto, Noriho Kamiya, Design of Lipid-Protein Conjugates Using Amphiphilic Peptide Substrates of Microbial Transglutaminase, ACS Applied Bio Materials, 10.1021/acsabm.8b00271, 1, 6, 1823-1829, 2018.12, Lipid modification of proteins plays a significant role in regulating the cellular environment. Mimicking natural lipidated proteins is a key technique for assessing the function of proteins modified with lipids and also to render self-assembly of lipids to a target protein. Herein, we report a facile method of conjugating proteins with lipid-fused peptides under homogeneous physiological conditions by using the microbial transglutaminase (MTG) reaction. MTG catalyzes the cross-linking reaction between a specific glutamine (Q) in a protein and a lysine (K) in newly designed lipid-fused peptides. The water-soluble peptide substrates for lipid modification, C14-X-MRHKGS, were newly synthesized, where C14, X, and MRHKGS represent myristic acid, linker peptides composed of G, P, or S, and MTG-reactive K surrounded with basic amino acids, respectively. The MTG-mediated cross-linking reaction between a protein fused with LLQG at the C-terminus and C14-X-MRHKGS (5 molar eq) dissolved in a phosphate saline solution resulted in lipid-protein conjugates with yields of 70 to 100%. The anchoring ability of the obtained lipid-protein conjugates to cell membranes was dependent on the number of G residues in the GnS linker, suggesting that self-assembly and hydrophobicity of the GnS motif serves to enhance membrane anchoring of lipid-protein conjugates.. |
| 41. |
Rahman Md Moshikur, Md Raihan Chowdhury, Rie Wakabayashi, Yoshiro Tahara, Muhammad Moniruzzaman, Masahiro Goto, Characterization and cytotoxicity evaluation of biocompatible amino acid esters used to convert salicylic acid into ionic liquids, International Journal of Pharmaceutics, 10.1016/j.ijpharm.2018.05.021, 546, 1-2, 31-38, 2018.07. |
| 42. |
Rie Wakabayashi, Masato Sakuragi, Shuto Kozaka, Yoshiro Tahara, Noriho Kamiya, Masahiro Goto, Solid-in-Oil Peptide Nanocarriers for Transcutaneous Cancer Vaccine Delivery against Melanoma, Mol. Pharm., 10.1021/acs.molpharmaceut.7b00894, 15, 3, 955-961, 2018.03. |
| 43. |
Rie Wakabayashi, Kensuke Yahiro, Kounosuke Hayashi, Masahiro Goto, Noriho Kamiya, Protein-grafted polymers prepared through a site-specific conjugation by microbial transglutaminase for an immunosorbent assay, Biomacromolecules, 18, 422-430, 2017.02. |
| 44. |
Yuya Hirakawa, Rie Wakabayashi, Ayaka Naritomi, Masato Sakuragi, Noriho Kamiya, Masahiro Goto, Transcutaneous immunization against cancer using solid-in-oil nanodispersions, Medicinal Chemical Communications, 6, 1387-1392, 2015.07. |
| 45. |
Rie Wakabayashi, Yuko Abe, Noriho Kamiya, Masahiro Goto, The Self-Assembly and Secondary Structure of Peptide Amphiphiles Determine the Membrane Permeation Activity, The Royal Society of Chemistry, 4, 30654-30657, 2014.07. |
| 46. |
Rie Wakabayashi, Ryutaro Ishiyama, Noriho Kamiya, Masahiro Goto, A Novel Surface-Coated Nanocarrier for Efficient Encapsulation and Delivery of Camptothecin to Cells, Medicinal Chemical Communications, 5, 1515-1519, 2014.07. |
| 47. |
Michiaki Kumagai, Shinya Shimoda, Rie Wakabayashi, Yumi Kunisawa, Takehiko Ishii, Kensuke Osada, Keiji Itaka, Nobuhiro Nishiyama, Kazunori Kataoka, Kenji Nakano, Effective transgene expression without toxicity by intraperitoneal administration of PEG-detachable polyplex micelles in mice with peritoneal dissemination, JOURNAL OF CONTROLLED RELEASE, 10.1016/j.jconrel.2012.03.021, 160, 3, 542-551, 2012.06, Block copolymer of poly(ethylene glycol)-block-poly{N-[N-(2-aminoethyl)-2-aminoethyl]aspartamide} (PEG-P[Asp(DET)]) has been originally introduced as a promising gene carrier by forming a nanomicelle with plasmid DNA. In this study, the polyplex micelle of PEG-SS-P[Asp(DET)], which disulfide linkage (SS) between PEG and cationic polymer can detach the surrounding PEG chains upon intracellular reduction, was firstly evaluated with respect to in vivo transduction efficiency and toxicity in comparison to that of PEG-P[Asp(DET)] in peritoneally disseminated cancer model. Intraperitoneal (i.p.) administration of PEG-SS-P[Asp(DET)] polyplex micelles showed a higher (P |
| 48. |
Gang Zhao, Rie Wakabayashi, Shinya Shimoda, Yumi Fukunaga, Michiaki Kumagai, Masao Tanaka, Kenji Nakano, Impaired activities of cyclic adenosine monophosphate-responsive element binding protein, protein kinase A and calcium-independent phospholipase A2 are involved in deteriorated regeneration of cirrhotic liver after partial hepatectomy in rats, HEPATOLOGY RESEARCH, 10.1111/j.1872-034X.2011.00868.x, 41, 11, 1110-1119, 2011.11, Aims: This study is to elucidate whether cyclic adenosine monophosphate (cAMP)-mediated signal is involved in lower regenerative potential of cirrhotic liver.. |
| 49. |
Rui Xu, Kenji Nakano, Hironori Iwasaki, Michiaki Kumagai, Rie Wakabayashi, Akio Yamasaki, Hiroyuki Suzuki, Ryuichi Mibu, Hideya Onishi, Mitsuo Katano, Dual blockade of phosphatidylinositol 3 '-kinase and mitogen-activated protein kinase pathways overcomes paclitaxel-resistance in colorectal cancer, CANCER LETTERS, 10.1016/j.canlet.2011.02.042, 306, 2, 151-160, 2011.07, Paclitaxel, one of key drugs to treat a wide range of malignancies, exhibits relative low sensitivity for colorectal cancer. The present study was to examine whether and how phosphatidylinositol 3'-kinase (PI3K) signals affect the sensitivity of colorectal cancer to paclitaxel. Four colorectal cancer cell lines were exposed to paclitaxel in the presence of PI3K signal inhibitors, such as LY294002, siRNA for Akt, or rapamycicn, with or without MAPK inhibitor, PD98059. Cell viability and apoptosis were determined by MTT assay, cell cycle analysis in flow cytometer and Hoechst nuclear staining. To analyze the PI3K activity, the expression in phosphorylated Akt and downstream effectors of p70S6 kinase (S6K) were evaluated by Western blot analysis. Paclitaxel alone (5-10 nM) did not induce the apoptosis in all four cell lines. Although LY294002 alone did not affect the cell viability, it suppressed the Akt and S6K activities and induced the sub-G1 arrest/apoptosis when paclitaxel was co-administered, as well as the Akt siRNA and rapamycin did. Simultaneous blockade of PI3K and MAPK pathways more suppressed the S6K activity and further increased the apoptosis. In conclusion, PI3K is involved in low susceptibility of colorectal cancer to paclitaxel and dual PI3K/MAPK targeting agents may evolve a new paclitaxel-based chemotherapy for colorectal cancer. (C) 2011 Elsevier Ireland Ltd. All rights reserved.. |
| 50. |
Soichiro Ogi, Tomohiro Ikeda, Rie Wakabayashi, Seiji Shinkai, Masayuki Takeuchi, Mechanically Interlocked Porphyrin Gears Propagating Two Different Rotational Frequencies, EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, 10.1002/ejoc.201001656, 10, 1831-1836, 2011.04, We report a new supramolecular rotor (1), in which a lanthanum( III) bis(porphyrinate)s double-decker (LaDD) bears two porphyrinatorhodium(III)-based rotors whose planes are situated orthogonally to one another. In [D-2]dichloromethane, H-1 NMR spectral results indicated that rotational information was transmitted from the LaDD rotor to two porphyrinatorhodium( III)-based rotors through mechanical interaction of the teeth of the rotors. Mechanical interaction was achieved among the multiple rotors in 1 even after switching the rotational activity of the LaDD rotor by using [D-5]pyridine. The spatial arrangement of the two porphyr inatorhodium(III)-based rotors is a critical factor in maintaining mechanical interaction between multiple molecular rotors when changing the dynamic properties of one rotor.. |
| 51. |
Rie Wakabayashi, Yohei Kubo, Kenji Kaneko, Masayuki Takeuchi, Seiji Shinkai, Olefin metathesis of the aligned assemblies of conjugated polymers constructed through supramolecular bundling, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 10.1021/ja063040x, 128, 27, 8744-8745, 2006.07. |
