Hidehiko Nakagawa

Nagoya City University, Nagoya, Aichi, Japan

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Publications (76)314.05 Total impact

  • [Show abstract] [Hide abstract]
    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; · 10.68 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; · 2.65 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 01/2014; · 2.84 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. 01/2014; 39:S5.
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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; · 6.38 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; · 5.44 Impact Factor
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    ABSTRACT: Histone N(epsilon)-methyl lysine demethylases KDM2/7 have been identified as potential targets for cancer therapies. Based on 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 μM, 0.2 μM, 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; · 5.61 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; · 10.68 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 07/2013; · 11.34 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; · 2.65 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 01/2013; 8(7):e68669. · 3.73 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; · 5.61 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; · 3.25 Impact Factor
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    ABSTRACT: Selective inhibitors of human sirtuin 2 (SIRT2), a deacetylase, are candidate therapeutic agents for neurodegenerative diseases such as Parkinson's disease and Huntington's disease as well as potential tools for elucidating the biological functions of SIRT2. On the basis of homology models of SIRT1 and SIRT2, we designed and prepared a series of 2-anilinobenzamide analogues. Enzyme assays using recombinant SIRT1 and SIRT2 revealed that 3'-phenethyloxy-2-anilinobenzamide analogues such as 33a and 33i are potent and selective SIRT2 inhibitors, showing more than 3.5-fold greater SIRT2-inhibitory activity and more than 35-fold greater SIRT2-selectivity compared with AGK2 (3), a previously reported SIRT2-selective inhibitor. Compound 33a also induced a dose-dependent selective increase of α-tubulin acetylation in human colon cancer HCT116 cells, indicating selective inhibition of SIRT2 in the cells. These 3'-phenethyloxy-2-anilinobenzamide derivatives represent an entry into a new class of SIRT2-selective inhibitors.
    Journal of Medicinal Chemistry 05/2012; 55(12):5760-73. · 5.61 Impact Factor
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    ABSTRACT: Nuclear oxidative stress damages genomic DNA and may lead to cell death, leading to aging and aging-related disorders. Though it is important to measure the nuclear oxidative stress separately, there are still little examples that applicable to living cells. We have designed and synthesized three bisbenzimide-nitroxides as probes to selectively visualize nuclear redox changes in terms of fluorescence. Compound 3, containing two radical moieties, showed the largest reduction-induced fluorescence change, with good localization in nuclei. RAW264.7 murine macrophage cells were loaded with compound 3 and then treated with 100μM hydrogen peroxide for 5min to show the fluorescence increase. This fluorescence increase was inhibited by pretreatment of 1mM ascorbic acid. These results show that compound 3 was suitable for nuclear-specific redox imaging in murine macrophages.
    Bioorganic & medicinal chemistry letters 03/2012; 22(5):1949-52. · 2.65 Impact Factor
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    ABSTRACT: We designed and synthesized a photocontrollable peroxynitrite (ONOO(-)) generator, P-NAP, which has N-methyl-N-nitrosoaminophenol structure with four methyl groups introduced onto the benzene ring to block reaction of the photodecomposition product with ONOO(-) and to lower the semiquinoneimine's redox potential. The semiquinoneimine intermediate generated by photoinduced release of nitric oxide (NO) reduces dissolved molecular oxygen to generate superoxide radical anion (O(2)(•-)), which reacts with NO to afford ONOO(-) under diffusion control (k = 6.7 × 10(9) M(-1) s(-1)). NO release from P-NAP under UV-A (330-380 nm) irradiation was confirmed by ESR spin trapping. Tyrosine nitration, characteristic of ONOO(-), was demonstrated by HPLC analysis of a photoirradiated aqueous solution of P-NAP and N-acetyl-l-tyrosine ethyl ester. ONOO(-) formation was confirmed with a ONOO(-)-specific fluorogenic probe, HKGreen-3, and compared with that from 3-(4-morpholinyl)sydnonimine hydrochloride (SIN-1), which is the most widely used ONOO(-) generator at present. The photoreaction of P-NAP was influenced by superoxide dismutase, indicating that generation of O(2)(•-) occurs before ONOO(-) formation. The quantum yield for formation of duroquinone, the main P-NAP photodecomposition product, was measured as 0.86 ± 0.07 at 334 nm with a potassium ferrioxalate actinometer. Generation of ONOO(-) from P-NAP in HCT-116 cells upon photoirradiation was successfully imaged with HKGreen-3A. This is the first example of a photocontrollable ONOO(-) donor applicable to cultured cells.
    Journal of the American Chemical Society 02/2012; 134(5):2563-8. · 10.68 Impact Factor
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    ABSTRACT: Nitroxyl (HNO), a one-electron-reduced form of nitric oxide, has various biological activities, including a cardioprotective effect. Here, we first synthesized another, more hydrophilic photocontrollable HNO donor (3), which can release HNO in a spatially and temporally controlled manner, and then examined the properties of our series of compounds as practical HNO donors in a cellular system under photocontrol. We selected compound 2 as the preferred donor, and used it to show that calcitonin gene-related peptide (CGRP) can be upregulated in A549 cells via photocontrolled HNO release. This result demonstrates the suitability of this photocontrollable HNO donor for biological investigations.
    Chemical & pharmaceutical bulletin 01/2012; 60(8):1055-62. · 1.70 Impact Factor
  • Kazuhiro Hishikawa, Hidehiko Nakagawa, Naoki Miyata
    ChemInform 09/2011; 42(37).
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    ABSTRACT: Photocontrollable ONOO(-) generation from a nitrobenzene derivative was demonstrated. The designed compound released NO in response to photoirradiation, and the resulting semiquinone reduced molecular oxygen to generate O(2)˙(-); reaction of the two generated ONOO(-), as confirmed with an ONOO(-) fluorescent probe, HKGreen-3.
    Chemical Communications 06/2011; 47(22):6449-51. · 6.38 Impact Factor
  • Kazuhiro Hishikawa, Hidehiko Nakagawa, Naoki Miyata
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    ABSTRACT: Nitric oxide (NO) is a key molecule in blood pressure regulation, neuromodulation, and biodefense. Since it is an unstable gas under ambient conditions, various NO donors have been developed and employed for biological studies as an alternative to direct NO application. However, many of them release NO via spontaneous decomposition, so that it is difficult to control the NO release. Therefore, NO donors from which NO release can be temporally and spatially well-controlled by means of photoexcitation offer considerable advantages. Various NO donors from which NO release is activated by photoirradiation have been reported. However, the maximum absorption of these NO donors is generally limited to the UV or short-wavelength visible light range, which does not adequately penetrate living tissues. To overcome this limitation, NO donors working via two-photon excitation (TPE) were developed by Ford and Prasad, based on NO release from Fe-nitrosyl complex. We have also developed TPE-type NO donors based on our photo-controllable 2,6-dimethylnitrobenzene derivatives, and investigated their NO-releasing properties. Here, we present an overview of photocontrollable NO donors and discuss their properties in relation to biological applications.
    Yakugaku zasshi journal of the Pharmaceutical Society of Japan 03/2011; 131(3):317-24. · 0.46 Impact Factor

Publication Stats

699 Citations
314.05 Total Impact Points

Institutions

  • 2005–2014
    • Nagoya City University
      • • Graduate School of Pharmaceutical Sciences
      • • Faculty of Pharmaceutical Sciences
      Nagoya, Aichi, Japan
  • 2012–2013
    • Kyoto Prefectural University of Medicine
      • Graduate School of Medical Science
      Kioto, Kyōto, Japan
  • 1998–2008
    • National Institute of Radiological Sciences
      • Research Center for Charged Particle Therapy
      Chiba-shi, Chiba-ken, Japan