Hidehiko Nakagawa

Nagoya City University, Nagoya, Aichi, Japan

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Publications (80)326.81 Total impact

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    ABSTRACT: Photocontrollable NO donors enable precise spatiotemporal release of NO under physiological conditions. We designed and synthesized a novel dimethylnitrobenzene-type NO donor, Flu-DNB-DB, which contains a carbon-carbon double bond in place of the amide bond of previously reported Flu-DNB. Flu-DNB-DB releases NO in response to one-photon activation in the blue wavelength region, and shows a greatly increased two-photon cross-section (δu) at 720nm (Flu-DNB: 0.12GM, Flu-DNB-DB: 0.98GM). We show that Flu-DNB-DB enables precisely controlled intracellular release of NO in response to 950nm pulse laser irradiation for as little as 1s. This near-infrared-light-controllable NO source should be a valuable tool for studies on the biological roles of NO. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.
    Bioorganic & medicinal chemistry letters 06/2015; 8(16). DOI:10.1016/j.bmcl.2015.05.095 · 2.33 Impact Factor
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    ABSTRACT: The levels of nitro fatty acids (NO2-FA), such as nitroarachidonic, nitrolinoleic, nitrooleic, and dinitrooleic acids, are elevated under various inflammatory conditions, and this results in different anti-inflammatory effects. However, other multiply nitrated and nitro-oxidized FAs have not been studied so far. Owing to the low concentrations in vivo, NO2-FA analytics usually relies on targeted gas chromatography-tandem mass spectrometry (MS/MS) or liquid chromatography-MS/MS, and thus require standard compounds for method development. To overcome this limitation and increase the number and diversity of analytes, we performed in-depth mass spectrometry (MS) profiling of nitration products formed in vitro by incubating fatty acids with NO2BF4, and ONOO(-). The modified fatty acids were used to develop a highly specific and sensitive multiple reaction monitoring LC-MS method for relative quantification of 42 different nitrated and oxidized species representing three different groups: singly nitrated, multiply nitrated, and nitro-oxidized fatty acids. The method was validated in in vitro nitration kinetic studies and in a cellular model of nitrosative stress. NO2-FA were quantified in lipid extracts from 3-morpholinosydnonimine-treated rat primary cardiomyocytes after 15, 30, and 70 min from stress onset. The relatively high levels of dinitrooleic, nitroarachidonic, hydroxynitrodocosapenataenoic, nitrodocosahexaenoic, hydroxynitrodocosahexaenoic, and dinitrodocosahexaenoic acids confirm the presence of multiply nitrated and nitro-oxidized fatty acids in biological systems for the first time. Thus, in vitro nitration was successfully used to establish a targeted LC-MS/MS method that was applied to complex biological samples for quantifying diverse NO2-FA. Graphical Abstract Schematic representation of study design which combined in vitro nitration of different fatty acids, MS/MS characterization and optimization of MRM method for relative quantification, which was applied to follow dynamic of fatty acid nitration in cellular model of SIN-1 treated cardiomyoctes.
    Analytical and Bioanalytical Chemistry 05/2015; DOI:10.1007/s00216-015-8766-3 · 3.58 Impact Factor
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    ABSTRACT: Hydrogen sulfide (H2S) has multiple physiological roles, for example, in vasodilation and inflammation. It is a highly reactive gas under ambient conditions, so controllable H2S donors are required for studying its biological functions. Here, we describe the design, synthesis and application of a H2S donor (SPD-2) that utilizes xanthone photochemistry to control H2S release. H2S generation from SPD-2 was completely dependent on UVA-irradiation (325-385nm), as confirmed by methylene blue assay and by the use of a H2S-selective fluorescent probe. SPD-2 was confirmed to provide controlled H2S delivery in live cells, and should be suitable for various biological applications. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.
    Bioorganic & Medicinal Chemistry Letters 12/2014; 25(2). DOI:10.1016/j.bmcl.2014.11.084 · 2.33 Impact Factor
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    ABSTRACT: Nitric oxide (NO) is a well-known free-radical molecule which is endogenously biosynthesised and shows various functions in mammals. To investigate NO functions, photocontrollable NO donors, compounds which release NO in response to light, are expected to be potentially useful. However, most of the conventional NO donors require harmful ultra-violet light for NO release. In this study, two dimethylnitrobenzene derivatives conjugated with coumarins were designed, synthesized and evaluated as photocontrollable NO donors. The optical properties and efficiency of photo-induced NO release were dependent upon the nature of the conjugation system. One of these compounds, Bhc-DNB (1), showed spatiotemporally well-controlled NO release in cultured cells upon exposure to light in the less-cytotoxic visible wavelength range (400–430 nm).
    Bioorganic & Medicinal Chemistry Letters 10/2014; 24(24). DOI:10.1016/j.bmcl.2014.10.075 · 2.33 Impact Factor
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    ABSTRACT: Lysine-specific demethylase 1 (LSD1), the first histone demethylase to be identified, catalyzes specifically the demethylation of the mono- and dimethyl groups of histone H3 lysine 4, and its dysregulation is thought to contribute to the development of cancer. We have recently reported that NCL1 (4) is the first cell-active LSD1-selective inhibitor. To find LSD1 inhibitors that show higher potency than NCL1 (4), we designed and synthesized an N-alkylated analogue of NCL1 (5), and evaluated its biological activity. In enzyme assays, compound 5 was six times more potent than 4, and compound 5 exhibited cell growth inhibition in cervical cancer HeLa cell line and neuroblastoma SH-SY5Y cell line. Compound 5 should be useful as a lead structure for anticancer drugs.
    Medicinal Chemistry Communication 09/2014; 6(3). DOI:10.1039/C4MD00330F · 2.63 Impact Factor
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    ABSTRACT: Hydrogen sulfide (H2S) has been recognized as one of the important gaseous cellular signaling molecules. H2S is considered to be involved in vascular relaxation, neurotransmission, and inflammation. To investigate the physiological and pharmacological actions of H2S, inorganic sulfide salts such as NaSH have been commonly used as H2S donors. However, these approaches suffer from an inability to precisely control the release rate and dosage. Although several H2S releasers have been reported, they are still incontrollable for H2S release except one example. To overcome these shortcomings, we focused on photochemical H2S generation. Controlled H2S release with photoirradiation has the potential to provide a high degree of control over release location, timing, and dosage. We envisaged that modification of H2S with suitable photolabile protecting groups would afford an H2S donor directly controllable with light, which would release H2S upon rapid photodissociation of the two protecting groups. A ketoprofenate photocage offers many advantages including good photochemical properties. In this study, we adopted ketoprofenate photocages for our new H2S photo-releaser, and developed a novel photocontrollable H2S releaser, which releases H2S proportionally to the photo-irradiation time and intensity. Photocontrolled H2S release from this compound was also demonstrated in biological bovine serum systems.
    Nitric Oxide 05/2014; 39:S5. DOI:10.1016/j.niox.2014.03.019 · 3.18 Impact Factor
  • Hidehiko Nakagawa, Naoki Fukushima, Naoya Ieda, Naoki Miyata
    Nitric Oxide 05/2014; 39:S48. DOI:10.1016/j.niox.2014.03.158 · 3.18 Impact Factor
  • Nitric Oxide 05/2014; 39:S48. DOI:10.1016/j.niox.2014.03.156 · 3.18 Impact Factor
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    ABSTRACT: Spatiotemporally controllable nitric oxide (NO)-releasers allow us to analyze the physiological effects of NO, a gaseous mediator that modulates many biological signaling networks, and are also candidate chemotherapeutic agents. We designed and synthesized a blue-light-controllable NO releaser, named NOBL-1, which bears an N-nitrosoaminophenol moiety for NO release tethered to a BODIPY dye moiety for harvesting blue light. Photoinduced electron transfer from N-nitrosoaniline to the antenna moiety upon irradiation with relatively noncytotoxic blue light (470-500 nm) should result in NO release with formation of a stable quinone moiety. NO release from NOBL-1 was confirmed by ESR spin trapping and fluorescence detection. Spatially controlled NO release in cells was observed with DAR-4M AM, a fluo-rogenic NO probe. We also demonstrated temporally controlled vasodilation of rat aorta ex vivo by blue-light-induced NO release from NOBL-1. This compound should be useful for precise examination of the functions of NO with excellent spatiotemporal control.
    Journal of the American Chemical Society 04/2014; 136(19). DOI:10.1021/ja5020053 · 11.44 Impact Factor
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    ABSTRACT: A series of 114 SIRT inhibitor candidates was assembled using 'click chemistry', by reacting two alkynes bearing 2-anilinobenzamide pharmacophore with 57 azide building blocks in the presence of Cu(I) catalyst. Screening identified two SIRT2-selective inhibitors, which were more SIRT2-selective than AGK2, a known SIRT2 inhibitor. These findings will be useful for further development of SIRT2-selective inhibitors.
    Bioorganic & medicinal chemistry letters 03/2014; 24(8). DOI:10.1016/j.bmcl.2014.03.026 · 2.33 Impact Factor
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    ABSTRACT: We recently discovered N-hydroxy-3-[1-(phenylthio)methyl-1H-1,2,3-triazol-4-yl]benzamide (NCC149) as a potent and selective histone deacetylase 8 (HDAC8) inhibitor from a 151-member triazole compound library using a click chemistry approach. In this work, we present a series of NCC149 derivatives bearing various aromatic linkers that were designed and synthesized as HDAC8-selective inhibitors. A series of in vitro assays were used to evaluate the newly synthesized compounds, four of which showed HDAC8 inhibitory activity similar to that of NCC149, and one of which displayed HDAC8 selectivity superior to that of NCC149. In addition, these top four compounds induced the increase of acetylated cohesin (an HDAC8 substrate) in HeLa cells in a dose-dependent manner, indicating inhibition of HDAC8 in the cells. While none of these compounds enhanced the acetylation of H3K9 (a substrate of HDAC1 and 2), only one compound refrained from increasing α-tubulin acetylation, a substrate of HDAC6, indicating that this compound is more selective for HDAC8 than the other derivatives. Furthermore, this HDAC8-selective inhibitor suppressed the growth of T-cell lymphoma cells more potently than did NCC149. These findings are useful for the further development of HDAC8-selective inhibitors.
    ChemMedChem 03/2014; 9(3). DOI:10.1002/cmdc.201300414 · 3.05 Impact Factor
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    ABSTRACT: We report the design, synthesis and application of a directly photocontrollable hydrogen sulfide (H2S) donor, which releases H2S proportionally to the intensity and duration of photoirradiation. Photocontrolled H2S release from this donor was also demonstrated in bovine serum. This H2S donor should be suitable for use in various biological systems.
    Chemical Communications 11/2013; 50(5). DOI:10.1039/c3cc47421f · 6.72 Impact Factor
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    ABSTRACT: Two-photon-excitation release of nitric oxide (NO) from our recently synthesized photo-labile NO donor, Flu-DNB, was confirmed to allow fine spatial and temporal control of NO release at the subcellular level in vitro. We then evaluated in vivo applications. Femtosecond near-infrared pulse laser irradiation of predefined regions of interest in living mouse brain treated with Flu-DNB induced NO-release-dependent, transient vasodilation specifically at the irradiated site. Photoirradiation in the absence of Flu-DNB had no effect. Further, NO release from Flu-DNB by pulse laser irradiation was shown to cause chemoattraction of microglial processes to the irradiated area in living mouse brain. To our knowledge, this is the first demonstration of induction of biological responses in vitro and in vivo by means of precisely controlled, two-photon-mediated release of NO.
    ACS Chemical Biology 08/2013; 8(11). DOI:10.1021/cb400361m · 5.36 Impact Factor
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    ABSTRACT: Histone Nε-methyl lysine demethylases KDM2/7 have been identified as potential targets for cancer therapies. On the basis of the crystal structure of KDM7B, we designed and prepared a series of hydroxamate analogues bearing an alkyl chain. Enzyme assays revealed that compound 9 potently inhibits KDM2A, KDM7A, and KDM7B, with IC50s of 6.8, 0.2, and 1.2 μM, respectively. While inhibitors of KDM4s did not show any effect on cancer cells tested, the KDM2/7-subfamily inhibitor 9 exerted antiproliferative activity, indicating the potential for KDM2/7 inhibitors as anticancer agents.
    Journal of Medicinal Chemistry 08/2013; 56(18). DOI:10.1021/jm400624b · 5.48 Impact Factor
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    ABSTRACT: Drug drop off: Given that lysine-specific demethylase 1 (LSD1) could be potently and selectively inactivated by delivering phenylcyclopropylamine (PCPA), a weak and nonselective LSD1 inhibitor, directly to the enzyme's active site, a novel series of LSD1 inactivators (1) were designed. Biological and mechanistic studies indicate that 1 inhibits LSD1 potently and selectively.
    Angewandte Chemie International Edition 08/2013; 52(33). DOI:10.1002/anie.201303999 · 11.34 Impact Factor
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    ABSTRACT: Nitroxyl (HNO) is a one-electron-reduced and protonated derivative of nitric oxide (NO), and has characteristic biological and pharmacological effects distinct from those of NO. However, studies of its biosynthesis and activities are restricted by the lack of versatile HNO detection methods applicable to living cells. Here, we report the first metal-free and reductant-resistant HNO imaging probe available for use in living cells, P-Rhod. It consists of a rhodol derivative moiety as the fluorophore, linked via an ester moiety to a diphenylphosphino-benzoyl group, which forms an aza-ylide upon reaction with HNO. Intramolecular attack of the aza-ylide on the ester carbonyl group releases a fluorescent rhodol derivative. P-Rhod showed high selectivity for HNO in the presence of various biologically relevant reductants, such as glutathione and ascorbate, in comparison with previous HNO probes. We showed that P-Rhod can detect not only HNO enzymatically gen-erated in the horseradish peroxidase-hydroxylamine system in vitro, but also intracellular HNO release from Angeli's salt in living cells. These results suggest that P-Rhod is suitable for detection of HNO in living cells.
    Journal of the American Chemical Society 07/2013; 135(34). DOI:10.1021/ja404757s · 11.44 Impact Factor
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    ABSTRACT: To find histone deacetylase 3 (HDAC3)-selective inhibitors, a series of 504 candidates was assembled using "click chemistry", by reacting nine alkynes bearing a zinc-binding group with 56 azide building blocks in the presence of Cu(I) catalyst. Screening of the 504-member triazole library against HDAC3 and other HDAC isozymes led to the identification of potent and selective HDAC3 inhibitors T247 and T326. These compounds showed potent HDAC3 inhibition with submicromolar IC50s, whereas they did not strongly inhibit other isozymes. Compounds T247 and T326 also induced a dose-dependent selective increase of NF-κB acetylation in human colon cancer HCT116 cells, indicating selective inhibition of HDAC3 in the cells. In addition, these HDAC3-selective inhibitors induced growth inhibition of cancer cells, and activated HIV gene expression in latent HIV-infected cells. These findings indicate that HDAC3-selective inhibitors are promising candidates for anticancer drugs and antiviral agents. This work also suggests the usefulness of the click chemistry approach to find isozyme-selective HDAC inhibitors.
    PLoS ONE 07/2013; 8(7):e68669. DOI:10.1371/journal.pone.0068669 · 3.53 Impact Factor
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    ABSTRACT: Recent studies have shown that nitroxyl (HNO) ((1)HNO/(3)NO(-)), which is the one-electron-reduced form of nitric oxide (NO), has unique biological activities, especially in the cardiovascular system, and HNO-releasing agents may have therapeutic potential. Since few HNO donors are available for use under physiological conditions, we synthesized and evaluated a series of Piloty's acid (PA) derivatives and evaluated their HNO-releasing activity under physiological conditions. N-Hydroxy-2-nitrobenzenesulfonamide (17) was the most efficient HNO donor among our synthesized PA derivatives, including the lead compound, 2-bromo-N-hydroxybenzenesulfonamide (2). The high HNO-releasing activity is suggested to be due to electronic and steric effects. Compound 17 may be a useful tool for biological experiments.
    Bioorganic & medicinal chemistry letters 02/2013; 23(8). DOI:10.1016/j.bmcl.2013.02.062 · 2.33 Impact Factor
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    ABSTRACT: To find HDAC8-selective inhibitors, we designed a library of HDAC inhibitor candidates, each containing a zinc-binding group that coordinates with the active-site zinc ion, linked via a triazole moiety to a capping structure that interacts with residues on the rim of the active site. These compounds were synthesized by using click chemistry. Screening identified HDAC8-selective inhibitors including C149 (IC50 = 0.070 μM), which was more potent than PCI-34058 (6) (IC50 = 0.31 μM), a known HDAC8 inhibitor. Molecular modeling suggested that the phenylthiomethyl group of C149 binds to a unique hydrophobic pocket of HDAC8, and the orientation of the phenylthiomethyl and hydroxamate moieties (fixed by the triazole moiety) is important for the potency and selectivity. The inhibitors caused selective acetylation of cohesin in cells and exerted growth-inhibitory effects on T-cell lymphoma and neuroblastoma cells (GI50 = 3-80 μM). These findings suggest that HDAC8-selective inhibitors have potential as anticancer agents.
    Journal of Medicinal Chemistry 11/2012; 55(22). DOI:10.1021/jm300837y · 5.48 Impact Factor
  • Hidehiko Nakagawa
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    ABSTRACT: Nitroxyl (HNO) is a small molecule with various pharmacological effects, including cardioprotective action. It is thought to serve as a modulator of various biochemical pathways. But, it is difficult to apply HNO directly for biological experiments or therapeutic treatment because it is highly reactive, readily dimerizing or reacting with biological targets under ambient conditions. Therefore, HNO donor molecules that release HNO under physiological conditions, especially those that allow precisely controllable release, would be useful to study the activities of HNO at the cellular level. This short review focuses on recently developed photocontrollable HNO-releasing compounds, which are expected to be suitable for achieving site-specific and temporally controlled HNO release in biomedical investigations. An illustrative application for the study of HNO-mediated upregulation of calcitonin gene-related peptide (CGRP) in A549 cells is described.
    Journal of inorganic biochemistry 10/2012; DOI:10.1016/j.jinorgbio.2012.10.004 · 3.27 Impact Factor

Publication Stats

1k Citations
326.81 Total Impact Points


  • 2005–2015
    • Nagoya City University
      • Graduate School of Pharmaceutical Sciences
      Nagoya, Aichi, Japan
  • 2011–2013
    • Japan Science and Technology Agency (JST)
      Edo, Tōkyō, Japan
  • 2007
    • RIKEN
      Вако, Saitama, Japan
    • Hiroshima International University
      Hirosima, Hiroshima, Japan
  • 1998–2004
    • National Institute of Radiological Sciences
      • Research Center for Charged Particle Therapy
      Tiba, Chiba, Japan