Hiroshi Terada, Satoru GOTO, Hitoshi Hori and Zenei Taira : QSAR and Drug Design: New Developments and Applications, (Pharmacochemistry Library Vol. 23) (Edited by T. Fujita), --- Structural Requirements of Leukotriene Antagonists ---, Elsevier Science Publishers, Amsterdam, Jan. 1995.
Hiroshi Terada, Satoru GOTO, Hitoshi Hori and Zenei Taira : Structural requirements of leukotriene antagonists, Elsevier, Amsterdam, Jan. 1995.
13.
Hitoshi Hori, Hideko Nagasawa and Hiroshi Terada : Environmental Oxidants, John Wiley & Sons, Inc., Sep. 1994.
(要約)
本書は, 生物環境や生体における酸素及び酸素ラジカルの反応とこれらがもたらす影響について論述した書である．(B5判 全630頁 編者:J.O.Nriagu, M.S.Simmon) Effects of Free Radicals from Hypoxic Cell Radiosensitizers, Hypoxic Cell Cytotoxins, and Bioreductive Anticancer Drugs on the Biological En-vironment.(425-443頁)において，生物環境下における酸素ラジカルの化学的挙動について，我々が分子設計した低酸素細胞放射線増感剤およびHypoxic cytotoxinに関する化学，生化学，薬理学，放射線生物学的研究成果を中心に関連抗がん剤の化成機構にも言及しながら考察した．
14.
Hitoshi Hori, Hideko Nagasawa and Hiroshi Terada : Advances in Environmental Sciences and Technology: Oxidants in the Environment (Edited by J.O.Nriagu), --- Effects of Oxygen Radicals From Hypoxic Cell Radiosensitizers and related Anticancer Drugs on the Biological Environment ---, John Wiley & Sons, Inc., 1994.
Shimamura Mariko, Yoshimi Ashino, Tsutomu Oikawa, Takao Iwaguchi, Hideko Nagasawa, Hitoshi Hori and Seiichi Inayama : Recent Advances in Chemotherapy, American Soci-ety for Micro-biology, 1994.
J.C. Powers, S. Odake, J. Oleksyszyn, Hitoshi Hori, T. Ueda, B. Boduszek and C.-M. Kam : Protease, Protease Inhibitors and Protease-Derived Peptides ( Agents and Actions Supplements Vol. 42) (Edited by J.C. Cheronis and J. E. Repine), --- Protease-Structures, Mechanism and Inhibitors ---, Birkhaeuser, 1993.
Hideko Nagasawa, Nasahiro Bando, Hitoshi Hori, Tetsuo Sato, Takuhito Tada, Yasuto Onoyama and Seiichi Inayama : Chemical Modifi-ers of Cancer Treatment, Pergamon Press, Apr. 1992.
(要約)
本書は，がんの放射線治療における化学修飾剤の開発研究に関する国際学会の重要な知見をまとめた成果報告集である．(編者:T. Wasserman, D. Siemann, P. Workman) 本編において，二官能基を持つ放射線増感剤ニトロイミダゾールヒドロキサム酸アナログの分子設計及び毒性，薬理活性について解析し，低酸素細胞放射線増感剤として優れていることを論述した．
18.
J.C. Powers, C.-M. Kan, Hitoshi Hori, J. Oleksyszyn and E.F. Jr. Meyer : Biochemistry of Pulmonary Emphysema (Edited by C. Grassi, J. Travis, L. Casali, & M. Luisetti), --- Synthetic Mechanism-Based and Transition-State Inhibitors for Human Neutrophil Elastase (pp.123-141) ---, Springer-Verlag, 1992.
Hideko Nagasawa, M. Bando, Hitoshi Hori, T. Satoh, T. Tada, Y. Onoyama and S. Inayama : Chemical Modifiers of Cancer Treatment (Edited by T. Wasserman, D. Siemann, P. Workman), --- Radiosensitizing, Toxicological, and Pharmacokinetic Properties of Hydroxamate Analogues of Nitroimidazoles as Bifunctional Radiosensitizers/Chemical Modifiers ---, Pergamon Press, 1992.
Hideko Nagasawa, Hitoshi Hori, Tetsuichi Shibata, Takashi Igarashi, Chieko Murayama, Tomoyuki Mori, Keisuke Sasai, Masaharu Takahashi, Mitsuyuki Abe, Haruyuki Fukuda, Yasuto Onoyama, Kazuo Sakaizumi, NObuharu Muta, Ichiro Kaneko and Seiichi Inayama : A Recent Acvance in The Radiosensitization of 2-Nitroimidazole-1-Acetohydroxamate Derivatives, Japan Radiosensitization Research Association, Kyoto, Oct. 1989.
(要約)
本書は癌の放射線治療における放射線増感効果についての第3回日中合同会議録を出版したものである．(B5判 全155頁) 本人担当部分:A Recent Acvance in The Radiosensitization of 2-Nitroimidazole-1-Acetohydroxamate De-rivatives.(82-92頁)において，我々の開発した新規低酸素性放射線増感剤に関して，in vitro及びin vivo活性試験を実施したところ，いずれもミソニダゾールよりも高い活性を示し，この結果，新しい放射線増感剤として注目されることを論述した．
25.
Hitoshi Hori, Chieko Murayama, Tomoyuki Mori, Yuta Shibamoto, Mitsuyuki Abe, Yasuto Onoyama and Seiichi Inayama : Chemical Modifiers of Cancer Treatment, --- 2-Nitroimidazole-1-acetohydroxamate as a Hypoxic Cell Radiosensitizer (pp.1029-1032) ---, Pergamon Press, Apr. 1989.
(要約)
本書は放射線によるがん治療における化学修飾物質についてのパリ会議での重要な知見をまとめたもので，我々は低酸素性放射線増感剤KIH-802についての分子設計，in vitro, in vivo活性さらに薬物動態の評価を行い既存のミソニダゾールより優れていることを明らかにした． 監修者 E.P.マレーズ，M.グイチャール，D.W. シーヤン
本書は癌の放射線治療のおける放射線増感効果にちいての第2回日中合同会議録を出版したもので，われわれは 新規放射線増感剤としての 2-ニトロイミダゾール-1-アセトヒドロキサメートKIH-802及びその誘導体の分子設計，合成及びin vitro,in vivo活性について述べ，これら薬剤がミソニダゾールより優れていることを論述した． 担当執筆部分の名称:A Recent Advance in the Radiosensitization of 2-Nitroimidazole-1-acetphydroxamate Derivatives. 担当部分 pp41-48
27.
C.Powers James, J.Harper Wada and Hitoshi Hori : Pulmonary Emphesema and Proteolysis: 1986, --- Mechanism-based Inhibitors of Human Leukocyte Elastase ---, Academic Press, 1987.
(要約)
本書は肺気腫及び蛋白質分解に関する1986年のトピックスを編集したもので,我々はヒト白血球エラスタ-ゼのメカニズム-ベイスド インヒビタ-特に,3-クロロイソクマリン,遷移状態インヒビタ-であるケトエステル及び動物実験についての我々の研究を中心に概説した．共同執筆者 James, C. Powers, J. Wade Harper, Hitoshi Hori 担当執筆部分の名称: Mechanism-based Inhibitors of Human Leukocyte Elastase. 担当部分 pp41-48
28.
Hitoshi Hori, Akira Yasutake, Yoshihiro Minematsu and C. James Powers : PEPTIDES: Structure & Function, --- 976Inhibition of human leukocyte elastase, porcine pancreatic elastase and catehpsin B by peptide ketones ---, Pierce Chemical Co, 1985.
T. Mori, C. Murayama, Hitoshi Hori and Seiichi Inayama : Radiosensitization of Hypoxic Cells Treated with Some Imidazole Derivatives, IAEA-TECDOC-266, 1982.
Kazuto Ohkura, Atsushi Tabata, Yoshihiro Uto and Hitoshi Hori : Molecular Interaction Between Boron Tracedrug UTX-51 Derivatives and Bovine Serum Albumin: Application to an Analytical Model of AGEs Destruction by Thermal Neutron Irradiation, Anticancer Research, Vol.42, No.8, 4017-4023, 2022.
(要約)
Boron tracedrugs possess global molecular tracking abilities and localized destructive power. We investigated the molecular properties of synthesized boron tracedrugs, including UTX-51, and their interactions with the advanced glycation end-product (AGE)-related protein bovine serum albumin (BSA). A conformational analysis of the compounds used in the present study was performed using CAChe (Fujitsu Inc., Tokyo, Japan) and the degree of stereo-hydrophobicity of the conformers obtained was verified using Mopac (Fujitsu Inc.). The interactive properties of global minimum conformers of the derivatives tested with BSA were assessed using Molegro Virtual Docker (CLC bio., Aarhus, Denmark). Among the compounds investigated, UTX-51 was confirmed to interact with BSA based on the formation of hydrogen bonds between BSA and UTX-51. UTX-51 is a promising boron tracedrug and can be used as the lead structure for developing a therapeutic agent for AGE-related diseases, including cancer.
Kazuto Ohkura, Atsushi Tabata, Yoshihiro Uto and Hitoshi Hori : Effect of Isomerization of TX-2036 Derivatives on the Interaction With Tyrosine Kinase Domain of EGF Receptor, Anticancer Research, Vol.40, No.8, 4675-4680, 2020.
(要約)
From the design and synthesis of enantiomers, we can expect to obtain two compounds with different pharmacokinetics and pharmacological activities at the same time, which is thought to lead to the development of efficient anticancer agents. Chiral-2-nitroimidazole TX-2036 derivatives exhibit stereo-configuration (R- and S-configuration)-dependent tyrosine kinase inhibitory activity, and the activity of the tyrosine kinase domain of EGF receptor (EGFR-tyk) is suppressed. In order to clarify the reason why the effects on EGFR-tyk activity differ depending on stereoisomers, we tried to analyze the interaction between each TX-2036 derivative and EGFR-tyk. The 2-nitroimidazole-based radiosensitizer TX-2036 series were synthesized and their molecular features were examined using protein kinase inhibition assay and molecular structural analysis. R-configured TXs (TX-2043, -2030, and -2036) exhibited more potent protein kinase inhibitory activity than S-configured TXs (TX-2044, - 2031, and -2037), and the IC value of TX-2036 was 1.8 μM. R-configured TXs interacted with Lys and Thr of EGFR-tyk. The combinations of amino acid residues targeted by the S-configured TXs were different from each other (Ile and Thr (TX-2044), Ser, Thr, and Thr (TX-2031), Gly, Cys, and Thr (TX-2037)). Preparing a series of isomers with different target sites was considered beneficial when the target was mutated.
Kazuto Ohkura, Atsushi Tabata, Yoshihiro Uto and Hitoshi Hori : Correlation Between Radiosensitizing Activity and the Stereo-structure of the TX-2036 Series of Molecules, Anticancer Research, Vol.39, No.8, 4479-4483, 2019.
(要約)
The stereo-configuration (R-, S-configuration) of chiral-2-nitroimidazole derivatives alters their radiosensitizing activity. This study aimed at examining the molecular features of these enantiomers by molecular simulation techniques. A series of 2-nitroimidazole-based radiosensitizer TX-2036 molecules were synthesized, and their profiles were examined using molecular structural analysis such as conformation analysis, molecular orbital analysis, and electrostatic potential analysis. R-configured TXs (TX-2043, -2030, -2036) had a weaker radiosensitizing activity than S-configured TXs (TX-2044, -2031, -2037), and R-compounds had a small minus electrostatic potential (ESP) field in the cyclopentene-1,3-dione region. S-configured TX-2046 had weaker radiosensitizing activity than R-configured TX-2045, and TX-2046 had a small minus ESP field as well as R-configured TX-2043, -2030, - 2036. The cyclopentene-1,3-dione involved in the small minus ESP field affected the radiosensitizing activity of the TX-2036 series of molecules.
Kazuto Ohkura, Y Kawaguchi, Y Tatematsu, Atsushi Tabata, Yoshihiro Uto and Hitoshi Hori : Structure-associated Functional Control of TX-1877 Series by Glyco-conjugation, Anticancer Research, Vol.38, No.7, 4241-4245, 2018.
(要約)
Sugar molecules are often used as a tool to structurally modify chemical compounds. The features of synthesized sugar-conjugated TX-1877 derivatives were herein examined. The molecular stabilities (reactivity) and hydrophobicities of sugar (e.g., monosaccharide and tetra-O-acetylated monosaccharide)-conjugated TXs were analyzed using a molecular simulation (e.g. molecular mechanics (MM) and molecular orbital (MO) analysis). The hydrophobicities of TX-1877 derivatives were increased by tetra-O-acetylation, and TX-2244 exhibited the most potent radiosensitizing activity (enhancement ratio: ER=2.30). The conformations and hydrophobicities of chemical compounds may be controlled by adding monosaccharide- and tetra-O-acetyl-conjugated sugars to TX-1877.
Kazuto Ohkura, Y Tatematsu, Y Kawaguchi, Yoshihiro Uto and Hitoshi Hori : Interactive Analysis of TX-1123 with Cyclo-oxygenase: Design of COX2 Selective TX Analogs., Anticancer Research, Vol.37, No.7, 3849-3854, 2017.
(要約)
To date, two cyclo-oxygenase (COX) isoforms, COX1 and COX2, have been identified. In the present study, the COX-inhibitory activities of TX-1123 derivatives with the 2-hydroxyarylidene-4-cyclopentene-1,3-dione structure were examined, and the binding profiles of TX-1123 to COXs were analyzed using docking simulations. X-Ray data on COX1 [protein data bank (PDB) ID=1PGG] and COX2 (PDB ID=3LN1) were used for molecular interactive simulations. The interactive profiles of TX-1123 derivatives with COXs were examined using a molecular simulation technique with Molegro Virtual Docker (CLC bio, Aarhus, Denmark). TX-1123 exhibited COX1-inhibitory activity [half-maximal-inhibitory concentration (IC)=1.57×10 M]. The COX2 inhibitory activity of TX-1123 was potent (IC=1.16×10 M), and the ratio of COX1/COX2 inhibition was 13.5. TX-1123 bound to the COX2 molecule, and the oxygen atom of the 4-cyclopentene-1,3-dione region of TX-1123 interacted with Cys and Gln of COX2. The TX-1123-binding pocket of COX2 differs from that of the COX2-selective celecoxib-binding pocket. TX-1123 exhibited a different COX2-interactive mechanism from that of celecoxib.
(キーワード)
Protein kinase / cyclo-oxygenase / kinase inhibitor
Eiji Nakata, Yoshihiro Yukimachi, Yoshihiro Uto, Hitoshi Hori and Takashi Morii : Latent pH-responsive ratiometric fluorescent cluster based on self-assembled photoactivated SNARF derivatives, Science and Technology of Advanced Materials, Vol.17, No.1, 431-436, 2016.
(要約)
We have developed a self-assembled fluorescent cluster comprising a seminaphthorhodafluor (SNARF) derivative protected by a photoremovable o-nitrobenzyl group. Prior to UV irradiation, a colorless and nonfluorescent cluster was spontaneously assembled in aqueous solution. After UV irradiation, the self-assembled cluster remained intact and showed a large enhancement in pH-responsive fluorescence. The unique pH responsive fluorescent cluster could be used as a dual-emissive ratiometric fluorescent pH probe not only in the test tube but also in HeLa cell cultures.
(キーワード)
Self-assembly / Self-organized materials / Sensors and actuators / Bio-inspired and biomedical materials / Self-assembly / fluorescent material / pH indicator / photoactivation / ratiometry
Yukio Morimoto, Hideko Nagasawa, Yoshihiro Uto, Toshiyuki Chatake and Hitoshi Hori : Structural Insight Into Protein Binding of Boron Tracedrug UTX-97 Revealed by the Co-Crystal Structure With Lysozyme at 1.26 Å Resolution, Journal of Pharmaceutical Sciences, Vol.105, No.8, 2298-2301, 2016.
(要約)
Boron neutron capture therapy (BNCT) is one of the numbers of radiotherapies for treatment of certain cancers. The ability of low-dose irradiation with neutrons or radioactive beams to provide an acceptable quality of life is an objective which has not yet been achieved; therefore it will be necessary to increase the efficiency of the neutron capture reaction by lower dose irradiation and by achieving higher drug concentrations in living cells. Drug selectivity in targeting the affected cellular compartment is most important. Molecular design and synthesis of drugs should be based on high resolution structures and analysis of specific compounds and host molecules; however, it is necessary to obtain crystals for X-ray structural analysis. Because compounds containing bulky functional groups are difficult to crystalize due to their flexibility, the method described here makes it possible to stabilize these compounds by complexing them with protein molecules. We have first solved the three-dimensional structure of a BNCT drug-protein molecule combination at 1.26 Å resolution, and discuss the nature of the interaction between a BNCT drug and the protein molecule residues.
Kazuto Ohkura, Yuki Kawaguchi, Y Tatematsu, Yoshihiro Uto and Hitoshi Hori : An Antitumor 2-Hydroxyarylidene-4-cyclopentene-1,3-Dione as a Protein Tyrosine Kinase Inhibitor: Interaction Between TX-1123 Derivatives and Src Kinase, Anticancer Research, Vol.36, No.7, 3645-3649, 2016.
(要約)
Protein tyrosine kinases (PTKs) play major roles in signal transduction during cell proliferation and apoptosis. Tyrphostin AG17 was previously shown to be a potent tumor growth inhibitor, while AG17 induced apoptosis and inhibited activity of cyclin-dependent kinase 2. We herein describe the binding features of tyrphostin AG17 analogs, such as TX-1123, with Src kinase (Src-K).MATERIALS AND METHODS:Structural data for Src-K were obtained from a protein data bank (ID=2SRC), and the molecular interactions between Src-K and TX-1123 derivatives were examined.RESULTS:TX-1123 exihibited potent Src-K inhibitory activity (half maximal-inhibitory concentration=2.2 M), and fit into the pocket of the Src-K molecule as well as c-AMP did.CONCLUSION:The binding profiles of TX-1123 derivatives differed from each other, while their Src-K inhibitory activities were affected by their fit in the Src-K molecule.
(キーワード)
Protein kinase / TX-1123 derivative / kinase inhibitor
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 27354635
R Munakata, T Inoue, T Koeduka, F Karamat, A Olry, A Sugiyama, K Takanashi, A Dugrand, Y Froelicher, R Tanaka, Yoshihiro Uto, Hitoshi Hori, J Azuma, A Hehn, F Bourgaud and K Yazaki : Molecular cloning and characterization of a geranyl diphosphate-specific aromatic prenyltransferase from lemon, Plant Physiology, Vol.166, No.1, 80-90, 2014.
(要約)
Prenyl residues confer divergent biological activities such as antipathogenic and antiherbivorous activities on phenolic compounds, including flavonoids, coumarins, and xanthones. To date, about 1,000 prenylated phenolics have been isolated, with these compounds containing various prenyl residues. However, all currently described plant prenyltransferases (PTs) have been shown specific for dimethylallyl diphosphate as the prenyl donor, while most of the complementary DNAs encoding these genes have been isolated from the Leguminosae. In this study, we describe the identification of a novel PT gene from lemon (Citrus limon), ClPT1, belonging to the homogentisate PT family. This gene encodes a PT that differs from other known PTs, including flavonoid-specific PTs, in polypeptide sequence. This membrane-bound enzyme was specific for geranyl diphosphate as the prenyl donor and coumarin as the prenyl acceptor. Moreover, the gene product was targeted to plastid in plant cells. To our knowledge, this is the novel aromatic PT specific to geranyl diphosphate from citrus species.
Masataka Oita, Yoshihiro Uto, Masahide Tominaga, Motoharu Sasaki, Yasuo Hara, Taro Kishi and Hitoshi Hori : Radiosensitivity Uncertainty Evaluation for the In Vitro Biophysical Modeling of EMT6 Cells, Anticancer Research, Vol.34, No.8, 4621-4626, 2014.
(要約)
The aims of this study were to evaluate the cell survival uncertainty distribution of radiation and to assess the accuracy of predictions of tumor response by using three different in vitro experimental cell cultures with radiosensitizers (including etanidazole). Using EMT6 cells and X-rays, the cell survival fraction was obtained from 15, 34, and 21 different experiments under normoxic, hypoxic, and hypoxic-plus-radiosensitizer culture, respectively. The α coefficients were 0.257 ± 0.188, 0.078 ± 0.080, and 0.182 ± 0.116 Gy(-1), respectively. The β coefficients were 0.0159 ± 0.0208, 0.0076 ± 0.0113, and 0.0062 ± 0.0077 Gy(-2), respectively. The α coefficient and the dose that killed half of the clonogens population (D50) were significantly different between normoxic cell and hypoxic cell cultures (p<0.01), respectively. The use of radiosensitizers under hypoxic conditions improved radiosensitivity. Our results suggest that parameter value distributions are required for biophysical modeling of applications for radiotherapy.
(キーワード)
Radiotherapy / biophysical modelling / in vitro / radiosensitivity uncertainty
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 25075109
Toshio Inui, Kaori Makita, Hirona Miura, Akiko Matsuda, Daisuke Kuchiike, Kentaro Kubo, Martin Mette, Yoshihiro Uto, Takahito Nishikata, Hitoshi Hori and Norihiro Sakamoto : Case Report: A Breast Cancer Patient Treated with GcMAF, Sonodynamic Therapy and Hormone Therapy, Anticancer Research, Vol.34, No.8, 4589-4593, 2014.
(要約)
Gc protein-derived macrophage-activating factor (GcMAF) occurs naturally in the human body. It has various functions, such as macrophage activation and antitumor activities. Recently, immunotherapy has become an attractive new strategy in the treatment of cancer. GcMAF-based immunotherapy can be combined with many other therapies. Sonodynamic therapy (SDT) using low-intensity ultrasound is a novel therapeutic modality. Ultrasound has been demonstrated to activate a number of sonosensitive agents allowing for the possibility of non-invasive targeted treatment for both superficial and deep-seated tumors. The current case study demonstrates that GcMAF and SDT can be used in combination with conventional therapies in patients with metastatic cancer, especially where treatment options are limited due to factors such as toxicity. This case study also suggests a new concept of cancer treatment using local destruction of cancer tissue, in this case conducted with SDT, to be used in combination with GcMAF immunotherapy as a systemic treatment.
Yoshihiro Uto, Dai Tamatani, Yusuke Mizuki, Yoshio Endo, Ikuo Nakanishi, Kei Ohkubo, Shunichi Fukuzumi, Masahiro Ishizuka, Toru Tanaka, Daisuke Kuchiike, Kentaro Kubo, Toshio Inui and Hitoshi Hori : Evaluation of the Sonosensitizing Activities of 5-Aminolevulinic Acid and Sn(IV) Chlorin e6 in Tumor-bearing Chick Embryos, Anticancer Research, Vol.34, No.8, 4583-4587, 2014.
(要約)
Recently, 5-aminolevulinic acid (5-ALA), precursors of protoporphyrin IX (PpIX), and Sn(IV) chlorin e6 (SnCe6) have been proposed as possible sonosensitizers for sonodynamic therapy of cancer. Therefore, we evaluated the pharmacokinetic properties and sonosensitizing activities of 5-ALA and SnCe6 in vivo by using the EMT6/KU tumor-bearing chick embryos. The concentration of PpIX in tumor and liver tissues and serum increased in a time-dependent manner after the i.v. administration of 5-ALA; PpIX concentrations reached their peak level after 5-7 h. The concentration of SnCe6 reached its maximum value in the tumor tissue and serum immediately after i.v. administration. The combined treatment of 5-ALA or SnCe6 with ultrasound irradiation showed a significant antitumor effect towards EMT6/KU solid tumors. We evaluated the pharmacokinetic properties and sonosensitizing activities of 5-ALA and SnCe6 in a chick embryo model and found that 5-ALA might be more suitable as a sonosensitizer than SnCe6.
Hitoshi Hori, Ryu Tada, Yoshihiro Uto, Eiji Nakata, Takashi Morii and Kai Masuda : A Neutron Dynamic Therapy with a Boron Tracedrug UTX-51 Using a Compact Neutron Generator, Anticancer Research, Vol.34, No.8, 4557-4560, 2014.
(要約)
We are developing a neutron dynamic therapy (NDT) with boron tracedrugs for a new mechanical-clearance treatment of pathotoxic misfolded, aggregated, and self-propagating prion-associated disease proteins. We present a compact neutron generator-based NDT using a boron tracedrug UTX-51. Our NDT is based on the weak thermal neutron-bombarded destructive action of UTX-51 on bovine serum albumin (BSA) using the neutron beams produced from a compact inertial electrostatic confinement fusion (IECF) neutron generator. BSA as an NDT molecular target was subjected to thermal neutron irradiation for eight hours using a compact neutron generator. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis pattern showed no protein band when 2 nmoles of BSA were irradiated with more than 100 nmoles of UTX-51, while BSA was not affected when irradiated without UTX-51. For the first time, we have succeeded in the molecular destruction of a prion-disease model protein, BSA, by NDT with a boron tracedrug, UTX-51, using a compact neutron generator.
Ryu Tada, Yoshihiro Uto, Shin-ichiro Masunaga, Yuko Kinashi, Koji Ono and Hitoshi Hori : An NDT Study of a Boron Tracedrug UTX-51 for Glycated BSA as an AGE Model, Anticancer Research, Vol.34, No.8, 4503-4507, 2014.
(要約)
Conventional therapies for diseases that are associated with protein aggregation typically prevent rather than clear protein aggregates. We have proposed neutron dynamic therapy (NDT) as a physical clearance therapy for protein aggregates. Advanced glycation end-products (AGEs), which are aggregated proteins, have been implicated in diabetes, Alzheimer's, and heart disease. Herein, we report the use of the boron tracedrug UTX-51, under thermal neutron irradiation, as an NDT for the targeted clearance of glycated bovine serum albumin (Gly-BSA), a model of AGEs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed to detect Gly-BSA decomposition by thermal neutron irradiation treated with UTX-51. The combination of UTX-51 with neutron irradiation showed a decrease in band intensity of Gly-BSA. We present our NDT strategy, which has been used for the targeted clearance of Gly-BSA, suggesting that NDT with boron tracedrugs can be used for the treatment of AGEs-related disease.
Chiaki Abe, Yoshihiro Uto, Ayaka Kawasaki, Chiho Noguchi, Ryo Tanaka, Toru Yoshitomi, Yukio Nagasaki, Yoshio Endo and Hitoshi Hori : Evaluation of the in vivo antioxidative activity of redox nanoparticles by using a developing chicken egg as an alternative animal model, Journal of Controlled Release, Vol.182, 67-72, 2014.
(要約)
Antioxidants have been demonstrated to exert beneficial effects as pharmacotherapies for cardiovascular diseases. The in vitro systems generally employed to evaluate antioxidants, however, are limited by having no appreciable in vivo redox status of the antioxidants. Therefore, we used our developing chicken egg model to evaluate the in vivo antioxidative activity of a redox nanoparticle possessing 2,2,6,6-tetramethylpiperidine-1-oxyl (RNP(O)). The 2,2'-azobis(2-methylpropionamidine) dihydrochloride (AAPH) elicited strong oxidative stress and its LD50 value for chick embryos was 3.5±0.9mg/egg. The low molecular weight nitroxide compound, 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL), which is known to have the highest level of antioxidant activity, showed no significant protective effect against AAPH-induced embryo lethality. On the contrary, RNP(O) had potent protective effects against AAPH-induced embryo lethality. Moreover, RNP(O) could significantly suppress the production of lipid peroxides in chick serum induced by hydrocortisone. Since RNP(O) has a longer retention time in blood than TEMPOL, RNP(O) may protect the embryo against lethal oxidative stress by suppressing lipid peroxidation. The validity of in vivo experiments using developing chicken eggs was supported by our data, where RNP(O) was determined to elicit strong antioxidative activity in vivo, irrespective of the lack of a significant difference in the in vitro activity between low-molecular weight TEMPOL and RNP(O). Our results support the use of the developing chicken egg model to evaluate the potential in vivo antioxidative activity of RNP(O).
Masataka Nagahama, Naoyuki Shimomura, Akito Nakagawa, Kenji Teranishi, Yoshihiro Uto and Hitoshi Hori : In Vivo Experimental Study of Nanosecond Pulsed Electric Field Effects on Solid Tumors, IEEE Transactions on Dielectrics and Electrical Insulation, Vol.20, No.4, 1266-1272, 2013.
Keiji Hirota, Yoshinori Nakagawa, Ryota Takeuchi, Yoshihiro Uto, Hitoshi Hori, Shinya Onizuka and Hiroshi Terada : Antitumor Effect of Degalactosylated Gc-Globulin on Orthotopic Grafted Lung Cancer in Mice, Anticancer Research, Vol.33, No.7, 2911-2915, 2013.
(要約)
Group-specific component (Gc)-globulin-derived macrophage-activating factor (GcMAF) generated by a cascade of catalytic reactions with deglycosidase enzymes exerts antitumor activity. We hypothesized that degalactosyl Gc-globulin (DG3), a precursor of GcMAF, also plays a role in recovery from cancer as well as GcMAF due to progression of deglycosylation by generally resident sialidases and mannosidases. We prepared the subtypes of DG3, such as 1f1f and 1s1s and its 22 homodimers, by using vitamin D3-binding Sepharose CL-6B and examined their antitumor activity in mice bearing Lewis lung carcinoma cells, by counting the number of nodules formed in their lungs. Antitumor activity of DG3 was observed regardless of its subtype, being equivalent to that of GcMAF. The injection route of DG3 affected its antitumor activity, with subcutaneous and intramuscular administration being more favorable than the intraperitoneal or intravenous route. In order to obtain significant antitumor activity, more than 160 ng/kg of DG3 were required. DG3 proved to be promising as an antitumor agent, similarly to GcMAF.
Toshio Inui, Daisuke Kuchiike, Kentaro Kubo, Martin Mette, Yoshihiro Uto, Hitoshi Hori and Norihiro Sakamoto : Clinical Experience of Integrative Cancer Immunotherapy with GcMAF, Anticancer Research, Vol.33, No.7, 2917-2920, 2013.
Daisuke Kuchiike, Yoshihiro Uto, Hirotaka Mukai, Noriko Ishiyama, Chiaki Abe, Daichi Tanaka, Tomohito Kawai, Kentaro Kubo, Martin Mette, Toshio Inui, Yoshio Endo and Hitoshi Hori : Degalactosylated/Desialylated Human Serum Containing GcMAF Induces Macrophage Phagocytic Activity and In Vivo Antitumor Activity, Anticancer Research, Vol.33, No.7, 2881-2885, 2013.
(要約)
The group-specific component protein-derived macrophage-activating factor (GcMAF) has various biological activities, such as macrophage activation and antitumor activity. Clinical trials of GcMAF have been carried out for metastatic breast cancer, prostate cancer, and metastatic colorectal cancer. In this study, despite the complicated purification process of GcMAF, we used enzymatically-treated human serum containing GcMAF with a considerable macrophage-stimulating activity and antitumor activity. We detected GcMAF in degalactosylated/desialylated human serum by western blotting using an anti-human Gc globulin antibody, and Helix pomatia agglutinin lectin. We also found that GcMAF-containing human serum significantly enhanced the phagocytic activity of mouse peritoneal macrophages and extended the survival time of mice bearing Ehrlich ascites tumors. We demonstrated that GcMAF-containing human serum can be used as a potential macrophage activator for cancer immunotherapy.
(キーワード)
Degalactosylated/desialylated human serum / GcMAF / macrophage phagocytic activity / in vivo antitumor activity
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 23780974
Junichi Morishige, Yoshihiro Uto, Hitoshi Hori, Kiyoshi Satouchi, Tanihiro Yoshiomoto and Akira Tokumura : Lysophosphatidic Acid Produced by Hen Egg White Lysophospholipase D Induces Vascular Development on Extraembryonic Membranes, Lipids, Vol.48, No.3, 251-262, 2013.
(要約)
Lysophosphatidic acid (lysoPtdOH), a lysophospholipid mediator, exerts diverse physiological effects, including angiogenesis, through its specific G-protein-coupled receptors. Previously, we showed that unfertilized hen egg white contains polyunsaturated fatty acid-rich lysoPtdOH and lysophospholipase D (lysoPLD). Here, we examined whether lysoPtdOH was produced by lysoPLD in the presence and absence of a hen fertilized ovum and what the physiological role of lysoPtdOH in hen egg white is. Mass spectrometry showed that fertilized hen egg white contained about 8 μM lysoPtdOH before incubation with an ovum, mainly comprised of 18:1- (12.6 %), 18:2- (37.8 %) and 20:4-molecular species (41.5 %). In an early gestation period, the lysoPtdOH was increased up to 9.6 μM, concomitant with a decrease in the level of polyunsaturated lysophosphatidylcholine (lysoPtdCho). Moreover, lysoPtdOH-degrading activities were found in egg white and the vitelline membrane, showing that these enzymes control lysoPtdOH levels in egg white. In an egg yolk angiogenesis assay, two lysoPtdOH receptor antagonists, Ki16425 and N-palmitoyl serine phosphoric acid (NASP), inhibited blood vessel formation induced by exogenously added 18:1-lysoPtdOH and its precursor lysoPtdCho on the hen yolk sac. Ki16425 and NASP also inhibited blood vessel formation in the chorioallantoic membrane (CAM). Furthermore, the relatively higher levels of LPA1, LPA2, LPA4 and LPA6 mRNA were present in the yolk sac and CAM. These results suggest that lysoPtdOH produced from lysoPtdCho by the action of lysoPLD in hen egg white is involved in the formation of blood vessel networks through several lysoPtdOH receptors on various extraembryonic membranes, including the yolk sac membrane and CAM.
Tomonori Kawashima, Sushma Manda, Yoshihiro Uto, Kei Ohkubo, Hitoshi Hori, Ken-ichiro Matsumoto, Kiyoshi Fukuhara, Nobuo Ikota, Shinya Onizuka, Shunichi Fukuzumi, Toshihiko Ozawa, Kazunori Anzai and Ikuo Nakanishi : Kinetics and Mechanism for the Scavenging Reaction of the 2,2-Diphenyl-1-picrylhydrazyl Radical by Synthetic Artepillin C Analogues, Bulletin of the Chemical Society of Japan, Vol.85, No.8, 877-883, 2012.
Kotaro Miyake, Masanori Nishioka, Satoru Imura, Erdenebulgan Batmunkh, Yoshihiro Uto, Hideko Nagasawa, Hitoshi Hori and Mitsuo Shimada : The novel hypoxic cytotoxin, TX-2098 has antitumor effect in pancreatic cancer; possible mechanism through inhibiting VEGF and hypoxia inducible factor-1 targeted gene expression, Experimental Cell Research, Vol.318, No.13, 1554-1563, 2012.
(要約)
Tumor hypoxia has been considered to be a potential therapeutic target, because hypoxia is a common feature of solid tumors and is associated with their malignant phenotype. In the present study, we investigated the antitumor effect of a novel hypoxic cytotoxin, 3-[2-hydroxyethyl(methyl)amino]-2-quinoxalinecarbonitrile 1,4-dioxide (TX-2098) in inhibiting the expression of hypoxia inducible factor-1α (HIF-1α), and consequently vascular endothelial cell growth factor (VEGF) expression in pancreatic cancer. The antitumor effects of TX-2098 under hypoxia were tested against various human pancreatic cancer cell lines using WST-8 assay. VEGF protein induced pancreatic cancer was determined on cell-free supernatant by ELISA. Moreover, nude mice bearing subcutaneously (s.c.) or orthotopically implanted human SUIT-2 were treated with TX-2098. Tumor volume, survival and expression of HIF-1 and associated molecules were evaluated in treatment versus control groups. In vitro, TX-2098 inhibited the proliferation of various pancreatic cancer cell lines. In s.c model, tumors from nude mice injected with pancreatic cancer cells and treated with TX-2098 showed significant reductions in volume (P<0.01 versus control). Quantitative real-time reverse transcription-PCR analysis revealed that TX-2098 significantly inhibited mRNA expression of the HIF-1 associated molecules, VEGF, glucose transporter 1 and Aldolase A (P<0.01 versus control). These treatments also prolong the survival in orthotopic models. These results suggest that the effect of TX-2098 in pancreatic cancer might be correlated with the expression of VEGF and HIF-1 targeted molecules.
Yoshihiro Uto, Syota Yamamoto, Hirotaka Mukai, Noriko Ishiyama, Ryota Takeuchi, Yoshinori Nakagawa, Keiji Hirota, Hiroshi Terada, Shinya Onizuka and Hitoshi Hori : Beta-Galactosidase treatment is a common first-stage modification of the three major subtypes of Gc protein to GcMAF, Anticancer Research, Vol.32, No.6, 2359-2364, 2012.
(要約)
The 1f1f subtype of the group-specific component (Gc) protein is converted into Gc protein-derived macrophage-activating factor (GcMAF) by enzymatic processing with β-galactosidase and sialidase. We previously demonstrated that preGc(1f1f)MAF, a full Gc(1f1f) protein otherwise lacking a galactosyl moiety, can be converted to GcMAF by treatment with mouse peritoneal fluid. Here, we investigated the effects of the β-galactosidase-treated 1s1s and 22 subtypes of Gc protein (preGc(1s1s)MAF and preGc22MAF) on the phagocytic activation of mouse peritoneal macrophages. We demonstrated the presence of Gal-GalNAc disaccharide sugar structures in the Gc(1s1s) protein by western blotting using peanut agglutinin and Helix pomatia agglutinin lectin. We also found that preGc(1s1s)MAF and preGc22MAF significantly enhanced the phagocytic activity of mouse peritoneal macrophages in the presence and absence of mouse peritoneal fluid. We demonstrate that preGc(1s1s)MAF and preGc22MAF proteins can be used as effective macrophage activators.
Kazuto Ohkura, Yuki Kawaguchi, Atsushi Tabata, Atsushi Yamamoto, Yasuo Shinohara, Hideaki Nagamune and Hitoshi Hori : Molecular Profiles of Cholesterol-dependent Cytolysin Family-derived 11mer Regions, Anticancer Research, Vol.32, No.6, 2343-2346, 2012.
(要約)
Cholesterol-dependent cytolysins (CDCs) are secreted from various types of bacteria and are involved in various diseases (e.g. abscess formation). Traditional CDCs has a conserved 11mer region, which is a key structure in membrane recognition. Based on the X-ray data of traditional CDC perfringolysin O (PFO), molecular models of intermedilysin (ILY), pyolysin (PLO), vaginolysin (VLY), and Streptococcus mitis-derived human platelet aggregation factor (Sm-hPAF) were constructed. The 11mer regions of these models were extracted, and their molecular features were analyzed. The dipole moments of these 11mer regions were classified into four types, and their stereo-hydrophobicity (dGW) was different. It was thought that these results influenced the species specificity and membrane recognition of each cytolysin. Traditional CDCs, ILY, PLO, and VLY consisted of four domains (domains 1 to 4). Domain 0 existed on the N-terminal side in Sm-hPAF in addition to these four domains. The 11mer sequence of Sm-hPAF is the same as that of VLY, but Sm-hPAF has slightly different characteristics (e.g. species specificity, membrane recognition, cholesterol dependency) compared to VLY. Dynamic structure analysis of domain 0 might clarify these differences.
Eiji Nakata, Masato Koizumi, Yohei Yamashita, Yoshihiro Uto and Hitoshi Hori : Boron tracedrug: design, synthesis, and pharmacological activity of phenolic BODIPY-containing antioxidants as traceable next-generation drug model, Advances in Experimental Medicine and Biology, Vol.737, 301-306, 2012.
Koichi Nonaka, Shinya Onizuka, Hiromi Ishibashi, Yoshihiro Uto, Hitoshi Hori, Toshiyuki Nakayama, Nariaki Matsuura, Takashi Kanematsu and Hikaru Fujioka : Vitamin D Binding Protein-Macrophage Activating Factor Inhibits HCC in SCID Mice, The Journal of Surgical Research, Vol.172, No.1, 116-122, 2012.
(要約)
A high incidence of recurrence after treatment is the most serious problem in hepatocellular carcinoma (HCC). Therefore, a new strategy for the treatment of the disease is needed. The aim of the present study was to investigate whether vitamin D binding protein-macrophage activating factor (DBP-maf) is able to inhibit the growth of HCC. The effects of DBP-maf on endothelial cells and macrophage were evaluated by WST-1 assay and phagocytosis assay, respectively. Human HCC cells (HepG2) were implanted into the dorsum of severe combined immunodeficiency (SCID) mice. These mice were divided into control and DBP-maf treatment groups (n = 10/group). The mice in the treatment group received 40 ng/kg/d of DBP-maf for 21 d. DBP-maf showed anti-proliferative activity against endothelial cells and also activated phagocytosis by macrophages. DBP-maf inhibited the growth of HCC cells (treatment group: 126 ± 18mm(3), untreated group: 1691.5 ± 546.9mm(3), P = 0.0077). Histologic examinations of the tumors revealed the microvessel density was reduced and more macrophage infiltration was demonstrated in the tumor of mice in the treatment group. DBP-maf has at least two novel functions, namely, an anti-angiogenic activity and tumor killing activity through the activation of macrophages. DBP-maf may therefore represent a new strategy for the treatment of HCC.
Kazuto Ohkura, Yasuo Shinohara and Hitoshi Hori : Efficiency of Antimicrobial Defense: Molecular Flexibility of Natural Defensin and Artificial Bis-quaternary Ammonium Compound, Anticancer Research, Vol.31, No.7, 2561-2564, 2011.
(要約)
Human α-defensins (such as HD5 and HD6) are typical bactericidal peptides. We examined the molecular features of HD5 and HD6 by molecular dynamics (MD) analysis. Molecular features of natural defensins and artificial bis-quaternary ammonium compounds (e.g. 1,6-polymethylenedithio)bis(1-octylpyridinium iodide: 4DTBP-m,8) were analyzed using molecular simulation techniques. HD5 and HD6 had different electrostatic potential profiles, which indicated the region-dependent hydrophobicity. 4DTBP-m,8 derivatives were significantly flexible, and many conformers existed. HD5 and HD6 indicated antimicrobial activity by restricted conformation.
Eiji Nakata, Masato Koizumi, Yohei Yamashita, Kenta Onaka, Yoshinori Sakurai, Natsuko Kondo, Koji Ono, Yoshihiro Uto and Hitoshi Hori : Design, Synthesis and Destructive Dynamic Effects of BODIPY-containing and Curcuminoid Boron Tracedrugs for Neutron Dynamic Therapy, Anticancer Research, Vol.31, No.7, 2477-2482, 2011.
(要約)
We previously designed the boron tracedrugs UTX-42, UTX-43, and UTX-44, which possess antioxidant potency. In order to explore their destructive dynamic effects when bombarded by weak thermal neutrons, we performed thermal neutron irradiation of bovine serum albumin (BSA) treated with the boron tracedrugs. Boron tracedrugs, including the boron dipyrromethene (BODIPY)-containing compounds UTX-42, UTX-44, and UTX-47 and the curcuminoid compounds UTX-50 and UTX-51, were designed for neutron dynamic therapy based on their molecular orbital calculation. Newly designed UTX-47, UTX-50, and UTX-51 were synthesized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to detect decomposition by thermal neutron irradiation of BSA treated with these boron tracedrugs. The combination of 1.0 μM BSA with 100 μM of each of the boron tracedrugs showed a decrease in band intensity after irradiation. All boron tracedrugs tested caused destructive dynamic damage of BSA during thermal neutron irradiation, suggesting that boron tracedrugs could be used as dynamic drugs for neutron dynamic therapy.
Yoshihiro Uto, Syota Yamamoto, Ryota Takeuchi, Yoshinori Nakagawa, Keiji Hirota, Hiroshi Terada, Shinya Onizuka, Eiji Nakata and Hitoshi Hori : Effect of the Gc-derived Macrophage-activating Factor Precursor (preGcMAF) on Phagocytic Activation of Mouse Peritoneal Macrophages, Anticancer Research, Vol.31, No.7, 2489-2492, 2011.
(要約)
The 1f1f subtype of the Gc protein (Gc(1f1f) protein) was converted into Gc-derived macrophage-activating factor (GcMAF) by enzymatic processing in the presence of β-galactosidase of an activated B-cell and sialidase of a T-cell. We hypothesized that preGc(1f1f)MAF, the only Gc(1f1f) protein lacking galactose, can be converted to GcMAF in vivo because sialic acid is cleaved by residual sialidase. Hence, we investigated the effect of preGc(1f1f)MAF on the phagocytic activation of mouse peritoneal macrophages. We examined the sugar moiety of preGc(1f1f)MAF with a Western blot using peanut agglutinin (PNA) and Helix pomatia agglutinin (HPA) lectin. We also found that preGc(1f1f)MAF significantly enhanced phagocytic activity in mouse peritoneal macrophages but only in the presence of the mouse peritoneal fluid; the level of phagocytic activity was the same as that observed for GcMAF. PreGc(1f1f)MAF can be used as an effective macrophage activator in vivo.
(キーワード)
Gc-Derived macrophage-activating factor precursor (preGcMAF) / 1f1f subtype of Gc protein (Gc1f1f protein) / phagocytic activity / mouse peritoneal macrophages
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 21873164
Chiaki Abe, Yoshihiro Uto, Takashi Nakae, Yuuya Shinmoto, Keiichiro Sano, Hiroko Nakata, Mizue Teraoka, Yoshio Endo, Hiroshi Maezawa, Shin-ichiro Masunaga, Eiji Nakata and Hitoshi Hori : Evaluation of the In vivo Radiosensitizing Activity of EtanidazoleUsing Tumor-bearing Chick Embryo, Journal of Radiation Research, Vol.52, No.2, 208-214, 2011.
(要約)
Chick embryos have been used as alternative experimental animals in various research fields, including virology, immunology, toxicology, oncology, and embryology. Until now, there have been no in vivo models using chick embryo to evaluate radiosensitizing activity. Here, the in vivo radiosensitizing activity of etanidazole, a well-known hypoxic cell radiosensitizer, was evaluated using tumor-bearing chick embryo. On the basis of tumor growth, drug administration and X-ray irradiation were performed on day 15 chick embryo, with the endpoint being day 18 chick embryo. In day 15 chick embryo, an X-ray irradiation dose of equal or less than 10 Gy did not cause significant tumor growth suppression. Intravenous administration of equal or less than 1.0 mg of etanidazole did not cause tumor growth suppression. Neither doses of equal or less than 8 Gy of irradiation nor 1.0 mg of etanidazole caused fatality of the chick embryo. On the basis of these results, we evaluated the radiosensitizing effect of a combination treatment with 8 Gy of irradiation and 1.0 mg of etanidazole. As noted above, 1.0 mg of etanidazole alone and 8 Gy of irradiation alone did not show tumor growth suppression. In contrast, a combination treatment with 8 Gy of irradiation and 1.0 mg of etanidazole showed 35% of significant tumor growth suppression. Thus, we succeeded in evaluating the in vivo radiosensitizing activity of etanidazole using tumor-bearing chick embryo. These results suggest that the use of tumor-bearing chick embryo may be part of a promising system for evaluating radiosensitizing activity.
(キーワード)
Tumor-bearing chick embryo / In vivo / Hypoxic cell radiosensitizer / Etanidazole / Radiosensitizing activity
Atsushi Ikeda, Yoshihiko Kawai, Jun-ichi Kikuchi, Motofusa Akiyama, Eiji Nakata, Yoshihiro Uto and Hitoshi Hori : Formation and regulation of fullerene-incorporation in liposomes under the phase transition temperature, Organic & Biomolecular Chemistry, Vol.9, No.8, 2622-2627, 2011.
(要約)
The fullerene-exchange reaction from a cyclodextrin cavity to liposomes represents one of the best methods to prepare lipid membrane-incorporated [70]fullerenes (C(70)). The C(70)-exchange reaction occurred completely at temperatures above the phase transition temperature (T(m)) of the liposomes; however, lowering the temperature to below the T(m) led to C(70) aggregation outside the liposomes. This observation has limited the development of more functional LMIC(70) using a variety of liposome compositions. In this paper, this reaction was found to occur efficiently by the addition of small amounts of lipids bearing a ϖ-moiety. The ϖ-moieties act as a gate when hydrophobic C(70) migrates into the hydrophilic liposome surface. Therefore, the ϖ-moieties should exist in the polar head groups of the lipids and the C(70)-exchange reaction can be controlled by pH.
Takeuchi Ryota, Yoshihiro Uto, Nakagawa Yoshinori, Hirota Keiji, Hiroshi Terada and Hitoshi Hori : A retrospective study of a calcium agent (E-Ca) using data on bone mineral density obtained by DXA method, Anticancer Research, Vol.30, No.8, 3199-3205, 2010.
(要約)
We performed a retrospective analysis of a calcium hydroxide-containing calcium agent (TACHIKAWA DENKAI CALCIUM™: E-Ca) based on data of bone mineral density obtained by dual-energy X-ray absorptiometry (DXA) to clarify the relationship between bone mineral density and E-Ca intake on an empty stomach in those who regularly use E-Ca. We found the percentage of volunteers with their age-matched (AM) values of above 100% to be 89%, and also a moderate positive correlation between AM values and the intake period of E-Ca. Our findings demonstrate that AM values can be used as an effective indicator assessing osteogenesis by regularly administrated calcium agents which exert their effects after long-term use.
(キーワード)
Absorptiometry, Photon / Adult / Aged / Bone Density / Calcium / Female / Humans / Male / Middle Aged / Retrospective Studies
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 20871041
Kazuto Ohkura, Katsumi Fukino, Yasuo Shinohara and Hitoshi Hori : N-Acetyl Transferase 2 Polymorphisms Associated with Isoniazid Pharmacodynamics: Molecular Features for Ligand Interaction, Anticancer Research, Vol.30, No.8, 3177-3180, 2010.
(要約)
Isoniazid is mainly metabolized by arylamine N-acetyltransferase 2 (NAT2). Rapid acetylator types have NAT2*4/*4 alleles. Intermediate acetylator types have any of the following alleles: NAT2*4/*5, *4/*6, or *4/*7. Slow acetylator types do not have the NAT2*4 allele. We examined molecular features of these NAT2 molecules. Structures of NAT2*5, *6, and *7 were constructed based on X-ray data of human NAT2*4 using a molecular modeling technique. The NAT2*4 molecule mostly occupied a positive electrostatic potential field. Ile(114) and Arg(197), which are mutation sites of NAT2*4 to NAT2*5 and NAT2*6, were located in the peripheral part of the positive field. Gly(286), the mutation site from NAT2*4 to NAT2*7, was located near coenzyme A (CoA) in the boundary of the positive and negative fields. Nonbinding energies between NAT2s and isoniazid were larger than those of CoA. Molecular polymorphism appears to influence the reactivity between NAT2 and the external ligand.
Eiji Nakata, Yoshihiro Yukimachi, Yoshijiro Nazumi, Yoshihiro Uto, Toshihiro Hashimoto, Yasuko Okamoto and Hitoshi Hori : Design of a SNARF-based Ratiometric Fluorescent Probe for Esterase, Chemistry Letters, Vol.39, No.7, 734-735, 2010.
(要約)
A novel ratiometric fluorescent probe for esterase has been developed based on the SNARF scaffold. The new SNARF derivative was used to monitor the activity of porcine liver esterase by ratiometric emission spectrum change.
Eiji Nakata, Yoshihiro Yukimachi, Yoshijiro Nazumi, Yoshihiro Uto, Hiroshi Maezawa, Toshihiro Hashimoto, Yasuko Okamoto and Hitoshi Hori : A newly designed cell-permeable SNARF derivative as an effective intracellular pH indicator., Chemical Communications, Vol.46, No.20, 3526-3528, 2010.
(要約)
We have successfully developed a new cell-permeable SNARF derivative that is activated inside the cell by ester hydrolysis and is notable for having reduced background fluorescence.
Nakashima Hitomi, Ikkyu Kazuhiro, Nakashima Kouichiro, Sano Keiichiro, Yoshihiro Uto, Eiji Nakata, Hideko Nagasawa, Sugimoto Hiroshi, Yoshitsugu Shiro, Nakagawa Yoshinori and Hitoshi Hori : Design of Novel Hypoxia-Targeting IDO Hybrid Inhibitors Conjugated with an Unsubstituted L-TRP as an IDO Affinity Moiety, Advances in Experimental Medicine and Biology, Vol.662, 415-421, 2010.
(要約)
We presented here design, syntheses and inhibitory activities of novel hypoxia-targeting IDO hybrid inhibitors conjugated with an unsubstituted L-Trp as an IDO affinity moiety without inhibitor 1MT, such as L-Trp-TPZ hybrids 1 (TX-2274), 2 (UTX-3), 3 (UTX-4), and 4 (UTX-2). TPZ-monoxide hybrids 1 and 3 were good competitive IDO inhibitors, while TPZ hybrids 2 and 4 were uncompetitive IDO inhibitors. Among them TPZ-monoxide hybrid 1 have the strongest IDO inhibitory activity. It suggests that TPZ-monoxide hybrids 1 and 3 are able to bind the active site of IDO, TPZ hybrids 2 and 4 are able to bind the enzyme-substrate complex. We proposed the possible mechanism of action of TPZ hybrid 2 that may first affect as a hypoxic cytotoxin, and then metabolized to TPZ-monoxide hybrid 1, which may do as an IDO inhibitor more effectively than its parent TPZ hybrid 2.
Douglas A. Kuntz, Shinichi Nakayama, Kayla Shea, Hitoshi Hori, Yoshihiro Uto, Hideko Nagasawa and David. R. Rose : Structural Investigation of the Binding of 5-Substituted Swainsonine Analogues to Golgi -Mannosidase II, ChemBioChem, Vol.11, No.5, 673-680, 2010.
(要約)
Golgi alpha-mannosidase II (GMII) is a key enzyme in the N-glycosylation pathway and is a potential target for cancer chemotherapy. The natural product swainsonine is a potent inhibitor of GMII. In this paper we characterize the binding of 5alpha-substituted swainsonine analogues to the soluble catalytic domain of Drosophila GMII by X-ray crystallography. These inhibitors enjoy an advantage over previously reported GMII inhibitors in that they did not significantly decrease the inhibitory potential of the swainsonine head-group. The phenyl groups of these analogues occupy a portion of the binding site not previously seen to be populated with either substrate analogues or other inhibitors and they form novel hydrophobic interactions. They displace a well-organized water cluster, but the presence of a C(10) carbonyl allows the reestablishment of important hydrogen bonds. Already approximately tenfold more active against the Golgi enzyme than the lysosomal enzyme, these inhibitors offer the potential of being extended into the N-acetylglucosamine binding site of GMII for the creation of even more potent and selective GMII inhibitors.
Kazuto Ohkura, Hitoshi Hori and Yasuo Shinohara : Role of C-terminal region of yeast ADP/ATP carrier 2 protein: dynamics of flexible C-terminal arm., Anticancer Research, Vol.29, No.11, 4897-4900, 2009.
(要約)
The ADP/ATP carrier catalyzes the exchange of ADP and ATP across the inner mitochondrial membrane. The molecular dynamics of modeled yeast type 2 AAC (yAAC2) was analyzed and molecular parameters were determined. The yAAC2 C-terminal moved flexibly and a negative electrostatic potential field (ESP) was located in the C-terminal region. The ESP field is always located in the C-terminal area during C-terminal truncation (d1-d9). Further C-terminal truncation occurred on field invagination into the core region (d11, d14, d16). The 2-6 C-terminal amino acid truncation did not affect the biological activity. The d7-d9 truncated mutants lost their biological function. A critical point in yAAC2 function was shown between d6 and d7 C-terminal truncation. The C-terminal structure of yAAC2 is thought to be involved in biological function control.
Kotaro Miyake, Mitsuo Shimada, Masanori Nishioka, Koji Sugimoto, Erdenebulgan Batmunkh, Yoshihiro Uto, Hideko Nagasawa and Hitoshi Hori : Downregulation of matrix metalloprotease-9 and urokinase plasminogen activator by TX-1877 results in decreased tumor growth and metastasis on xenograft model of rectal cancer, Cancer Chemotherapy and Pharmacology, Vol.64, No.5, 885-892, 2009.
(要約)
Purpose It is well known that hypoxic milieu is the primary cancer environment. Therefore, tumor hypoxia is considered to be a potential therapeutic target. In the present study, we investigated the antitumor and antimetastatic effect of hypoxic cell radiosensitizer, TX-1877 on xenograft model of rectal cancer. Methods Nude mice bearing subcutaneously or orthotopically implanted human colon cancer cell lines HCT-116 and HT-29 were treated with TX-1877, irradiation or TX-1877 with irradiation. Tumor volume, survival, expression of matrix metalloproteinase (MMP)-2, MMP-7, MMP-9 and urokinase-type plasminogen activator (uPA) and incidence of lymph node metastasis were evaluated in treatment versus control group. Results In subcutaneous model, tumor treated with TX-1877 and irradiation showed significant reductions in volume (P < 0.05 vs. control, TX-1877 or irradiation group). Quantitative real-time reverse transcription-PCR and immunohistochemical analysis revealed that TX-1877 significantly inhibited expression of the MMP-9 and uPA. These treatments also inhibited the para-aortic lymph node metastasis, however, did not prolong the survival in orthotopic model. Conclusions These data show that the treatment of TX-1877 with irradiation decreased growth of human rectal cancer and, furthermore, suppressed lymph node metastasis.
Eiji Nakata, Yukimachi Yoshihiro, Kariyazono Hirokazu, Im Seongwang, Abe Chiaki, Yoshihiro Uto, Hiroshi Maezawa, Hashimoto Toshihiro, Okamoto Yasuko and Hitoshi Hori : Design of a bioreductively-activated fluorescent pH probe for tumor hypoxia imaging, Bioorganic & Medicinal Chemistry, Vol.17, No.19, 6952-6958, 2009.
(要約)
We have designed and evaluated UTX-12 as a novel fluorescent pH probe for tumor hypoxia imaging. UTX-12 consists of a p-nitro benzyl moiety, which is a latent hypoxia-selective leaving group activated by nitro reduction, directly linked to SNARF. Although UTX-12 itself is colorless and non-fluorescent in aqueous solution, nitro reduction triggers the release of SNARF which has well-characterized long wavelength absorption and fluorescence that is sensitive to pH. The resultant SNARF, released intracellularly by enzymatic reduction of UTX-12, allows measurement of pH by pH-dependent dual emission shifts. UTX-12 showed clear differences in fluorescence behavior between hypoxic and aerobic conditions in liver microsomes and inside V79 cells. These data are confirmation that UTX-12 is biologically reduced inside tumor cells and the released SNARF should monitor intracellular pH of tumor cells selectively with reduced background signal.
Mok-Ryeon Ahn, Kazuhiro Kunimasa, Shigenori Kumazawa, Tsutomu Nakayama, Kazuhiko Kaji, Yoshihiro Uto, Hitoshi Hori, Hideko Nagasawa and Toshiro Ohta : Correlation between antiangiogenic activity and antioxidant activity of various components from propolis, Molecular Nutrition & Food Research, Vol.53, No.5, 643-651, 2009.
(要約)
Propolis possesses various physiological activities. In this study, we examined the antiangiogenic and antioxidant activities of various components from propolis: acacetin, apigenin, artepillin C, caffeic acid phenethyl ester, chrysin, p-coumaric acid, galangin, kaempferol, pinocembrin, and quercetin. The effects of these components were tested on in vitro models of angiogenesis, tube formation and growth of human umbilical vein endothelial cells (HUVECs). Furthermore, these components were evaluated for their antioxidant activities by 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging and ferric reducing/antioxidant power (FRAP) assays. Two propolis components, caffeic acid phenethyl ester, and quercetin, possessed strong inhibitory effects on tube formation and on endothelial cell proliferation and, coincidentally, showed strong antioxidant activity. Artepillin C, galangin, and kaempferol also possessed strong antiangiogenic and antioxidant activities to a slightly less degree. In contrast, acacetin, apigenin, and pinocembrin possessed a considerable degree of antiangiogenic activities, although they showed very low antioxidant activities. From these results, we propose that components from propolis such as artepillin C, caffeic acid phenethyl ester, galangin, kaempferol, and quercetin might represent a new class of dietary-derived antioxidative compounds with antiangiogenic activities. These propolis components may have the potential to be developed into pharmaceutical drugs for the treatment of angiogenesis-dependent human diseases such as tumors.
Shanta M. Messerli, Mok-Ryeon Ahn, Kazuhiro Kunimasa, Miyako Yanagihara, Tomoki Tatefuji, Ken Hashimoto, Victor Mautner, Yoshihiro Uto, Hitoshi Hori, Shigenori Kumazawa, Kazuhiko Kaji, Toshiro Ohta and Hiroshi Maruta : Artepillin C (ARC) in Brazilian green propolis selectively blocks oncogenic PAK1 signaling and suppresses the growth of NF tumors in mice, Phytotherapy Research, Vol.23, No.3, 423-427, 2009.
(要約)
There are mainly three types of propolis whose major anticancer ingredients are entirely different: (1) CAPE (caffeic acid phenethyl ester)-based propolis in Europe, Far East and New Zealand, (2) artepillin C (ARC)-based Brazilian green propolis and (3) Brazilian red propolis. It was shown previously that NF (neurofibromatosis)-associated tumors require the kinase PAK1 for their growth, and CAPE-based propolis extracts such as Bio 30 suppress completely the growth of NF tumors in vivo by blocking PAK1 signaling. Also it was demonstrated that ARC suppresses angiogenesis, suggesting the possibility that ARC also blocks oncogenic PAK1 signaling. Here it is shown for the first time that both ARC and green propolis extract (GPE) indeed block the PAK1 signaling selectively, without affecting another kinase known as AKT. Furthermore, it was confirmed that ARC as well as GPE suppress almost completely the growth of human NF tumor xenografts in mice, as does Bio 30. These results suggest that both CAPE-based and ARC-based propolis extracts are natural anti-PAK1 remedies and could be among the first effective NF therapeutics available on the market. Since more than 70% of human cancers such as breast and prostate cancers require the kinase PAK1 for their growth, it is quite possible that GPE could be potentially useful for the treatment of these cancers, as is Bio 30.
(キーワード)
artepillin C / Brazilian green propolis / PAK1 / neurofibromatosis
Kazuto Ohkura, Yuki Kawguchi, Yasuo Watanabe, Yasuhiro Masubuchi, Yasuo Shinohara and Hitoshi Hori : Flexible Structure of Cytochrome P450: Promiscuity of Ligand Binding in the CYP3A4 Heme Pocket, Anticancer Research, Vol.29, No.3, 935-942, 2009.
(要約)
CYP3A4 is the most abundant xenobiotic-metabolizing cytochrome P450 isoform. We examined the structural features of the CYP3A4 molecule with regard to ligand access. The deleted amino acid sequences of X-ray data sets of CYP3A4s were complemented by molecular modeling techniques. Molecular features of the ligand accessible regions in CYP3A4 were analyzed and their molecular parameters (e.g. dipole moment, solvation free energy, electrostatic potential fields) were determined. Three ligand accessible regions (region 1-3) were present in erythromycin-bound CYP3A4, and these dipole moments indicated the same features as ketoconazole- or metyrapone-bound CYP3A4 molecules. In progesterone-bound CYP3A4, four candidate ligand accessible regions were observed and progesterone could be bound by two selected ligand accessible regions. The heme pocket of CYP3A4 is very flexible and is able to interact with various types of substrate.
Yoshihiro Uto, Daisuke Koyama, Mamoru Otsuki, Naoki Otomo, Tadashi Shirai, Chiaki Abe, Eiji Nakata, Hideko Nagasawa and Hitoshi Hori : A chemical biosynthesis design for an antiatherosclerosis drug by acyclic tocopherol intermediate analogue based on "isoprenomics", Advances in Experimental Medicine and Biology, Vol.645, 109-114, 2009.
(要約)
Phytyl quinols, namely acyclic tocopherols, are key intermediates of tocopherol biosynthesis, but their biological activities remain unclear. We therefore investigated the structure-activity relationship of phytyl quinols to apply a chemical biosynthesis design for an antiatherosclerosis drug based on isoprenomics. We have achieved the biosynthesis-oriented design and synthesis of alpha- (TX-2254) and beta-(TX-2247) phytyl quinol as an unnatural intermediate, other gamma- (TX-2242) and delta-(TX-2231) phytyl quinol as a natural one. Geometry optimization and Molecular orbital (MO) calculation of TX-2254 showed a unique right-angle structure; however, MO energy of TX-2254 and d-alpha-tocopherol were very similar. Radical reactivity of TX-2231 was equal to dl-alpha-tocopherol, whereas TX-2254, TX-2247, and TX-2231 showed lower reactivity than dl-alpha-tocopherol. All four phytyl quinols showed almost the same moderate inhibitory activity against low-density lipoprotein (LDL) oxidation instead of their different degree of C-methylation with character different from tocopherols. In vivo toxicities of phytyl quinols against chick embryo chorioallantoic membrane (CAM) vasculature were hardly observed. We proposed phytyl quinols were possible antioxidants in plants and animals, like vitamin E.
Yoshio Takeuchi, Tomoya Fujiwara, Yoshihito Shimone, Hideki Miyataka, Toshio Satoh, Kenneth L. Kirk and Hitoshi Hori : Possible involvement of radical intermediates in the inhibition of cysteine proteases by allenyl esters and amides, Bioorganic & Medicinal Chemistry Letters, Vol.18, No.23, 6202-6205, 2008.
(要約)
In order to investigate crystallographically the mechanism of inhibition of cysteine protease by alpha-methyl-gamma,gamma-diphenylallenecarboxylic acid ethyl ester 3, a cysteine protease inhibitor having in vivo stability, we synthesized N-(alpha-methyl-gamma,gamma-diphenylallenecarbonyl)-L-phenylalanine ethyl ester 4. Reaction of 4 with thiophenol, the SH group of which has similar pK(a) value to that of cysteine protease, produced oxygen-mediated radical adducts 6 and 7 in ambient air but did not proceed under oxygen-free conditions. Catalytic activities of two thiol enzymes including cathepsin B were also lowered in the absence of oxygen. These results suggest that cysteine protease can act through an oxygen-dependent radical mechanism.
Kotaro Miyake, Mitsuo Shimada, Masanori Nishioka, Koji Sugimoto, Erdenebulgan Batmunkh, Yoshihiro Uto, Hideko Nagasawa and Hitoshi Hori : The novel hypoxic cell radiosensitizer, TX-1877 has antitumor activity through suppression of angiogenesis and inhibits liver metastasis on xenograft model of pancreatic cancer., Cancer Letters, Vol.272, No.2, 325-335, 2008.
(要約)
Tumor hypoxia has been considered to be a potential therapeutic target, because hypoxia is a common feature of solid tumors and is associated with their malignant phenotype. In the present study, we investigated the antitumor effect of hypoxic cell radiosensitizer, TX-1877 in inhibiting angiogenesis and liver metastasis on pancreatic cancer xenograft model. The antitumor effects of TX-1877 were tested against various human tumor cell lines using cell proliferation assay. Nude mice bearing s.c. or orthotopically implanted human SUIT-2 were treated with TX-1877 alone, irradiation alone or TX-1877 and irradiation. Tumor volume, survival, expression of angiogenic molecules and liver metastasis were evaluated in treatment versus control groups. In vitro, TX-1877 inhibited the proliferation and potentiated the radiosensitivity of various pancreatic cancer cell lines. In an orthotopic model, tumors from nude mice injected with pancreatic cancer cells and treated with TX-1877 and irradiation showed significant reductions in volume (p<0.05 versus control, TX-1877 alone or irradiation alone). Quantitative real-time reverse transcription-PCR and immunohistochemical analysis revealed that treatment with TX-1877 alone or with TX-1877 and irradiation inhibited expression of the angiogenic molecules, vascular endothelial growth factor; basic fibroblast growth factor, interleukin-8 and matrix metalloproteinase 9 more than control or did treatment with irradiation alone. These treatments also induced apoptosis in cancer cells. These data show that treatment of TX-1877 and irradiation decreased growth of human pancreatic cancer, suppressed angiogenesis and inhibited liver metastasis, leading to prolonged survival.
Hitomi Nakashima, Yoshihiro Uto, Eiji Nakata, Hideko Nagasawa, Kazuhiro Ikkyu, Noriko Hiraoka, Kouichiro Nakashima, Yuki Sasaki, Hiroshi Sugimoto, Yoshitsugu Shiro, Toshihiro Hashimoto, Yasuko Okamoto, Yoshinori Asakawa and Hitoshi Hori : Synthesis and biological activity of 1-methyl-tryptophan-tirapazamine hybrids as hypoxia-targeting indoleamine 2,3-dioxygenase inhibitors, Bioorganic & Medicinal Chemistry, Vol.16, No.18, 8661-8669, 2008.
(要約)
We have designed and synthesized new hypoxic-neoplastic cells-targeted indoleamine 2,3-dioxygenase (IDO) inhibitors. 1-Methyl-tryptophan (1MT)-tirapazamine (TPZ, 3-amino-1,2,4-benzotriazine 1,4-dioxide) hybrid inhibitors including 1 (TX-2236), 2 (TX-2235), 3 (TX-2228), and 4 (TX-2234) were prepared. All of these compounds were uncompetitive IDO inhibitors. TPZ-monoxide hybrids 1 and 3 showed higher IDO inhibitory activities than TPZ hybrids 2 and 4. Among these hybrids, hybrid 1 was the most potent IDO inhibitor. TPZ hybrids 2 and 4 showed stronger hypoxia-selective cytotoxicity than TPZ to EMT6/KU cells. These data suggest that TPZ hybrids 2 and 4 may act through their dual biological functions: first, they function as hypoxic cytotoxins in hypoxic cells, and then are metabolized to their TPZ-monoxide (3-amino-1,2,4-benzotriazine 1-oxide) hybrids, which function as IDO inhibitors.
Shinichi Nakayama, Yoshihiro Uto, Kanako Tanimoto, Yasuhiro Okuno, Yuki Sasaki, Hideko Nagasawa, Eiji Nakata, Ken Arai, Kaori Momose, Tetsuro Fujita, Toshihiro Hashimoto, Yasuko Okamoto, Yoshinori Asakawa, Satoru GOTO and Hitoshi Hori : TX-2152: A conformationally rigid and electron-rich diyne analogue of FTY720 with in vivo antiangiogenic activity, Bioorganic & Medicinal Chemistry, Vol.16, No.16, 7705-7714, 2008.
(要約)
We designed FTY720 analogues with conformationally rigid and electron-rich acetylenic chains as antiangiogenic agents (the monoyne 1: TX-2148, the diyne 2: TX-2152, the triyne 3: TX-2256). Molecular orbital (MO) calculations of our designed acetylenic analogues and FTY720 showed that the localization of the lowest unoccupied MO and the highest occupied MO increased from phenyl ring to acetylenic chain compared with that of FTY720. These acetylenic analogues were synthesized from p-hydroxyphenylethanol as a starting material. The construction of the acetylenic chain was carried out by an iterative strategy using a Sonogashira cross-coupling reaction and desilylative bromination in two steps. The corresponding overall yields of the monoyne 1, the diyne 2, and the triyne 3 were 27% (11 steps), 13% (13 steps), and 10% (15 steps). The in vivo antiangiogenic activities of these acetylenic analogues and FTY720 were evaluated by the chick embryo chorioallantoic membrane (CAM) assay and compared to the activities of the known antiangiogenic agent TNP-470. The diyne 2 showed more potent antiangiogenic activity (90% inhibition) than FTY720 (77% inhibition) and other acetylenic analogues (the monoyne 1: 42% inhibition, the triyne 3: 60% inhibition), and TNP-470 (82% inhibition) at a dose of 10 μg/CAM, without showing toxicity. The diyne 2 also had potent inhibitory activity at a dose of 5 and 2.5 μg/CAM. These results indicate that the flexibility of C8 alkyl chain of FTY720 is not required for its antiangiogenic activity. We suggest that the diyne 2 (TX-2152) may be a promising candidate as an antiangiogenic agent for antineoplastic drug discovery.
We designed chiral 2-nitroimidazole derivatives containing a 2-aminomethylene-4-cyclopentene-1,3-dione moiety as antiangiogenic hypoxic cell radiosensitizers. Based on results of molecular orbital calculations, the 2-aminomethylene-4-cyclopentene-1,3-dione moiety is expected to show high electrophilicity comparable to that of the 2-methylene-4-cyclopentene-1,3-dione moiety included in TX-1123 and tyrphostin AG17. We evaluated the antiangiogenic and radiosensitizing effects of the new compounds, along with other biological properties including their activities as hypoxic cytotoxicities and protein tyrosine kinase (PTK) inhibitory activities. Among the compounds tested, 5 (TX-2036) proved to be the strongest antiangiogenic hypoxic cell radiosensitizer. All the other chiral 2-nitroimidazole derivatives having 2-aminomethylene-4-cyclopentene-1,3-dione moiety tested were also antiangiogenic hypoxic cell radiosensitizers. The PTK inhibitory activity of 5 (TX-2036) showed this to be a promising and potent EGFR kinase inhibitor, having an IC50 value of lower than 2 μM. This compound also was an Flt-1 kinase inhibitor having an IC50 value of lower than 20 μM. Our results show that these chiral 2-nitroimidazole derivatives that contain the 2-aminomethylene-4-cyclopentene-1,3-dione moiety as a potent antiangiogenic pharmacophoric descriptor are promising lead candidates for the development of antiangiogenic hypoxic cell radiosensitizers.
Niraldo Paulino, Lemos Sheila Rago Abreu, Yoshihiro Uto, Daisuke Koyama, Hideko Nagasawa, Hitoshi Hori, Verena M. Dirsch, Angelika M. Vollmar, Amarilis Scremin and Walter A. Bretz : Anti-inflammatory effects of a bioavailable compound, Artepillin C, in Brazilian propolis, European Journal of Pharmacology, Vol.587, No.1-3, 296-301, 2008.
(要約)
Artepillin C is the major compound in the Brazilian green propolis from Baccharis dracunculifolia. Our aim in this study was to investigate the anti-inflammatory effects, absorption, and bioavailability of Artepillin C in mice. The animals used were male Swiss mice subjected to: paw oedema by carrageenan (300 μg/paw), carrageenan-induced peritonitis, and prostaglandin E2 determination. We also measured in vitro nitric oxide production by RAW 264.7 cells and NF-κB activity in HEK 293 cells. Finally, we measured the absorption and bioavailability of Artepillin C in plasma from mice by means of GC-MS after a single oral dose (10 mg/kg). In vivo, Artepillin C produced a maximal inhibition of 38% after 360 min on paw oedema. Artepillin C also decreased the number of neutrophils during peritonitis (IC50: 0.9 (0.5 1.4) mg/kg). Treatment with Artepillin C decreased prostaglandin E2 by 29 ± 3% and 58 ± 5% at 1 and 10 mg/kg, respectively, with a mean ID50 of 8.5 (8.0 8.7)mg/kg). Similarly, in in vitro models, Artepillin C (3, 10, or 100 μM) decreased nitric oxide production by RAW 264.7 cells with a mean IC50 of 8.5 (7.8 9.2) μM. In HEK 293 cells, Artepillin C reduced NF-κB activity with a mean IC50 of 26 (22 30) μg/ml), suggesting anti-inflammatory activity, particularly during acute inflammation. Lastly, Artepillin C was absorbed after an oral dose (10 mg/kg) with maximal peaks found at 1 h (22 μg/ml). Collectively, Artepillin C showed anti-inflammatory effects mediated, at least in part, by prostaglandin E2 and nitric oxide inhibition through NF-κB modulation, and exhibited bioavailability by oral administration.
Takashi Nakae, Yoshihiro Uto, Motoko Tanaka, Haruna Shibata, Eiji Nakata, Masahide Tominaga, Hiroshi Maezawa, Toshihiro Hashimoto, Kenneth L. Kirk, Hideko Nagasawa and Hitoshi Hori : Design, synthesis, and radiosensitizing activities of sugar-hybrid hypoxic cell radiosensitizers, Bioorganic & Medicinal Chemistry, Vol.16, No.2, 675-682, 2008.
(要約)
We have designed sugar-hybrid TX-1877 derivatives conjugated with sugar moieties including β-glucose (β-Glc), β-galactose (β-Gal), α-mannose (α-Man) and N-acetyl-β-galactosamine (β-GalNAc). Compound 1 (TX-1877) was glycosylated with appropriate peracetylated sugars using BF3-OEt2 to give acetylated sugar-hybrids, 5 (TX-2244), 6 (TX-2245), 7 (TX-2246), and 10 (TX-2243). Removal of the acetyl groups afforded the sugar-hybrids having free hydroxyl groups, 11 (TX-2141), 12 (TX-2218), 13 (TX-2217) and 14 (TX-2068). We evaluated their radiosensitizing activities by an in vitro radiosensitization assay. All free hydroxyl hybrids have lower enhancement ratio (ER) values (ER 1.43) and lower n-octanol/water partition coefficient (Poct) values (Poct < 1.00 × 10-2) than does 1 (TX-1877, ER = 1.75, Poct: 5.60 × 10-2). All acetylated hybrids have similar Poct values (3.55 × 10-2 1.05 × 10-1) to 1 (TX-1877) and have improved ER values (ER 1.47) compared to the hybrids having free hydroxyl groups. Among these, 5 (TX-2244) is the most active radiosensitizer (ER = 2.30). We found a good correlation (r = 0.866) between the magnitude of Poct (logPoct) and the ER value of 5 (TX-2244), 6 (TX-2245), 7 (TX-2246), 10 (TX-2243) and 1 (TX-1877), suggesting that increasing the hydrophobicity is reflected in increased in vitro radiosensitizing activity. In the present study, we have succeeded in producing sugar-hybrid hypoxic cell radiosensitizers that have an increased radiosensitizing activity that does not depend on increased hydrophobicity.
(キーワード)
Hypoxic cell radiosensitizers / Sugar-hybrids / Electronic states / Hydrophobicity
Kazuto Ohkura, Yoshihiro Uto, Hideko Nagasawa and Hitoshi Hori : Effect of molecular chirality and side chain bulkiness on angiogenesis of haloacetylcarbamoyl-2-nitroimidazole compounds, Anticancer Research, Vol.27, No.6A, 3693-3700, 2007.
(要約)
BACKGROUND: Angiogenesis is required for tumor growth and metastasis, and is an exciting target for cancer treatment. We designed and synthesized antiangiogenetic TX agent (TX-1898, -1900), and analyzed their structural features. TXs have a chiral center and S- and R-enantiomers. Conformation analysis and molecular dynamics simulation were undertaken. Materials and METHODS: Molecular models of TXs were constructed using InsightlI-Discover. Conformation analysis was performed with CONFLEX, and z-matrix data were extracted to calculate molecular orbital (MO) parameters (i.e. solvation free energy (dGW)). Their molecular dynamics were simulated with the Discover3 module, and the total energy and dihedral angles were estimated. RESULTS: The methyl-including TXs (Group 1: TX-1863, -1878, -1866, -1879) had 130-229 conformers (-1.26-14.6 kcal/mol). The t-butyl-including group (Group 2: TX-1880, -1881, -1882, -1883) had 244 - 294 conformers (3.69 - 16. 76 kcal/mol), and the p-t-butylphenyl-containing TXs (Group 3: TX-1897, -1899, -1898, -1900) had 584 - 711 conformers (-7.48 -5.18 kcal/mol). The dGWprofile of nine samples, which were extracted from these conformers, were examined and one minimum dGW point was observed in the haloacetylcarbamoyl-2-nitroimidazole TXs (Group 1 -3). CONCLUSION: TX-1898 exhibited significant antiangiogenic activity. The order of antiangiogenic activity was as follows: TX-1898 (93% at 5 microg/pellet) > TX-1900 (82% at 5 microg/pellet) > TX-1897 (64% at 10 microg/pellet) > TX-1899 (58% at 10 microg/pellet). The chiral center has an important role for orienting the molecular characteristics.
Haruhiko Sakuraba, Kazunari Yoneda, Kumiko Yoshihara, Kyoko Satoh, Ryushi Kawakami, Yoshihiro Uto, Hideaki Tsuge, Katsuyuki Takahashi, Hitoshi Hori and Toshihisa Ohshima : Sequential aldol condensation catalyzed by hyperthermophilic 2-deoxy-D-ribose-5-phosphate aldolase., Applied and Environmental Microbiology, Vol.73, No.22, 7427-7434, 2007.
(要約)
Genes encoding 2-deoxy-d-ribose-5-phosphate aldolase (DERA) homologues from two hyperthermophiles, the archaeon Pyrobaculum aerophilum and the bacterium Thermotoga maritima, were expressed individually in Escherichia coli, after which the structures and activities of the enzymes produced were characterized and compared with those of E. coli DERA. To our surprise, the two hyperthermophilic DERAs showed much greater catalysis of sequential aldol condensation using three acetaldehydes as substrates than the E. coli enzyme, even at a low temperature (25 degrees C), although both enzymes showed much less 2-deoxy-d-ribose-5-phosphate synthetic activity. Both the enzymes were highly resistant to high concentrations of acetaldehyde and retained about 50% of their initial activities after a 20-h exposure to 300 mM acetaldehyde at 25 degrees C, whereas the E. coli DERA was almost completely inactivated after a 2-h exposure under the same conditions. The structure of the P. aerophilum DERA was determined by X-ray crystallography to a resolution of 2.0 A. The main chain coordinate of the P. aerophilum enzyme monomer was quite similar to those of the T. maritima and E. coli enzymes, whose crystal structures have already been solved. However, the quaternary structure of the hyperthermophilic enzymes was totally different from that of the E. coli DERA. The areas of the subunit-subunit interface in the dimer of the hyperthermophilic enzymes are much larger than that of the E. coli enzyme. This promotes the formation of the unique dimeric structure and strengthens the hydrophobic intersubunit interactions. These structural features are considered responsible for the extremely high stability of the hyperthermophilic DERAs.
Shin-ichiro Masunaga, Hideko Nagasawa, Yoshihiro Uto, Hitoshi Hori, Kenji Nagata, Minoru Suzuki, G Kashino, Yuko Kinashi and Koji Ono : The usefulness of mild temperature hyperthermia combined with continuous tirapazamine administration under reduced dose-rate irradiation with gamma-rays, International Journal of Hyperthermia, Vol.23, No.1, 29-35, 2007.
(要約)
We clarified the usefulness of mild temperature hyperthermia (MTH) in combination with the continuous administration of tirapazamine (TPZ) under reduced dose-rate irradiation (RDRI) using gamma-rays. SCC VII tumour-bearing mice received a continuous administration of 5-bromo-2'-deoxyuridine (BrdU) to label all proliferating (P) cells. Then, they received a 24 h continuous subcutaneous infusion of TPZ either with or without MTH under high dose-rate irradiation (HDRI) or RDRI using gamma-rays. After the irradiation, the tumour cells were isolated and incubated with a cytokinesis blocker, and the micronucleus (MN) frequency in non-proliferating tumour cells without BrdU labeling (= quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. The MN frequency in the total tumour cell populations was determined using tumours that were not pretreated with BrdU. The sensitivity of both the total and Q cell populations, especially the latter, was significantly reduced with RDRI compared with HDRI. TPZ increased the sensitivity of both populations, with a slightly more remarkable increase in Q cells. Further, MTH combined with TPZ raised the sensitivity of both the total and Q cell populations, especially the latter, under RDRI more markedly than under HDRI. From the viewpoint of solid tumour control as a whole, including intratumour Q-cell control, the use of TPZ, especially in combination with MTH, is useful for suppressing the reduction in the sensitivity of tumour cells caused by the decrease in irradiation dose rate in vivo.
Shin-ichiro Masunaga, Hideko Nagasawa, Kenji Nagata, Minoru Suzuki, Yoshihiro Uto, Hitoshi Hori, Yuko Kinashi and Koji Ono : Dependency of the effect of a vascular disrupting agent on sensitivity to tirapazamine and gamma-ray irradiation upon the timing of its administration and tumor size, with reference to the effect on intratumor quiescent cells, Journal of Cancer Research and Clinical Oncology, Vol.133, No.1, 47-55, 2007.
(要約)
The effect of vascular disrupting agent ZD6126 with time on the sensitivity to the hypoxic cytotoxin tirapazamine (TPZ) and gamma-rays was examined in large and small solid tumors. Mice bearing SCC VII tumors 1 or 1.5 cm in diameter received 5-bromo-2'-deoxyuridine (BrdU) continuously to label all proliferating (P) cells, followed by injection with or without ZD6126. In the absence of ZD6126, or 1 or 24 h following ZD6126 injection, the response to TPZ or gamma-ray irradiation in quiescent (Q) cells was assessed in terms of induced micronucleus (MN) frequency using immunofluorescence staining for BrdU. The MN frequency in the total cell population was determined from the tumors not pretreated with BrdU. Another group of tumor-bearing mice received a series of test doses of gamma-rays while alive or after tumor clamping to obtain hypoxic fractions (HFs) in the tumors. One hour after ZD6126 injection, both small and large tumors showed lower and higher sensitivity, and 24 h after, higher and lower sensitivity, to gamma-rays and TPZ, respectively, than the tumors not treated with ZD6126. Further, they showed larger and smaller HFs 1 and 24 h after ZD6126 injection, respectively. Without ZD6126 and 1 h after injection, small tumors were more sensitive to gamma-rays and less sensitive to TPZ than large tumors, probably due to the smaller HFs than large tumors. In contrast, 24 h after the injection, these differences in sensitivity and the HF between small and large tumors were reversed. The changes in sensitivity and the size of the HF were more marked in the total cell population than in Q cells. Following ZD6126 treatment, in terms of tumor control, especially large tumors and total tumor cell population, administering TPZ 1 h later and gamma-ray irradiation 24 h later were effective. Intratumor physiologic factors such as the size of the HF, depending on the time after ZD6126 injection, have to be taken into account when combining another treatment with ZD6126.
Shin-ichiro Masunaga, Yoshinori Sakurai, Kenji Nagata, Minoru Suzuki, Akira Maruhashi, Yuko Kinashi, Hideko Nagasawa, Yoshihiro Uto, Hitoshi Hori and Koji Ono : The usefulness of a continuous administration of tirapazamine combined with reduced dose-rate irradiation using gamma-rays or reactor thermal neutrons, The British Journal of Radiology, Vol.79, No.948, 991-998, 2006.
(要約)
We clarified the usefulness of the continuous administration of tirapazamine (TPZ) in combination with reduced dose-rate irradiation (RDRI) using gamma-rays or reactor thermal neutrons. Squamous cell carcinoma (SCC) VII tumour-bearing mice received a continuous administration of 5-bromo-2'-deoxyuridine (BrdU) to label all proliferating (P) cells. Then, they received a single intraperitoneal injection or 24 h continuous subcutaneous infusion of TPZ in combination with conventional dose-rate irradiation (CDRI) or RDRI using gamma-rays or thermal neutrons. After irradiation, the tumour cells were isolated and incubated with a cytokinesis blocker, and the micronucleus (MN) frequency in cells without BrdU labelling ( = quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. The MN frequency in the total tumour cells was determined using tumours that were not pre-treated with BrdU. The sensitivity of both total and Q cells, especially of Q cells, was significantly reduced with RDRI compared with CDRI. Combination of TPZ increased the sensitivity of both populations, with a slightly more remarkable increase in Q cells. Furthermore, the continuous administration of TPZ raised the sensitivity of both total and Q cell populations, especially the former, more markedly than the single administration, whether combined with CDRI or RDRI using gamma-rays or thermal neutrons. From the viewpoint of solid tumour control as a whole, including intratumour Q-cell control, the use of TPZ, especially when administered continuously, combined with RDRI, is useful for suppressing the reduction in the sensitivity of tumour cells caused by the decrease in irradiation dose rate in vivo.
Kazuto Ohkura, Hideko Nagasawa, Yoshihiro Uto, Natsuko Okamura, Aya Murakami and Hitoshi Hori : The Role of Gc Protein Oligosaccharide Structure as a Risk Factor for COPD, Anticancer Research, Vol.26, No.6A, 4073-4078, 2006.
(要約)
The risk of chronic obstructive pulmonary disease (COPD) is related to Gc protein allele type, such as Gc*1F, Gc*IS, Gc*2. It has been reported that Gc*1F increased COPD risk, while Gc*2 suppressed the risk. Thus, the allele type of Gc protein is an important factor in COPD. These Gc proteins differ in sugar composition at Thr418 or Thr420. In this study, features of the sugar structure of modeled Gc proteins were investigated. Gc protein (GclF, Gc1S, Gc2) models were constructed based on X-ray data of vitamin D binding protein (ID=1J7E) using InsightII-Discover with the Homology module, and the molecular orbital (MO) parameters [e.g., dipole moment, solvation free energy (dGW)] of the oligosaccharide were analyzed. The MO parameter of the sugar moiety was different for each Gc protein model. In beta-1,4 bond models, the dipole moment of Gc2 protein was larger (56.6 debye) than Gcl type (GclF: 21.9, Gc1S: 29.8 debye) protein, and it was directed towards the intermolecular space. The Gc2 oligosaccharide region was the most hydrophobic (dGW=-999.4 KI) among the Gc proteins analyzed in this study. The electrostatic potential (ESP) field of beta-1,4 type Gc2 protein was similarly distributed to beta-1,4 linked Gcl-type proteins (GclF, GclS). In the beta-1,3 type Gc protein models, the results of these parameters (i.e., dipole moment, dGW and ESP) were similar to those of beta-1,4 type models. Conclusion: The relationship between COPD risk and the features of the sugar structure in Gc proteins was examined, and it appeared that the active factors (i.e., dipole moment, dGW) might be risk factors for COPD, but passive factors (i.e., ESP) did not affect COPD risk. The bond type (beta-1,4 or beta-1,3) between galactose and N-acetylgalactosamine did not affect the molecular features.
Atsushi Miyoshi, Yoshihiko Kitajima, Takao Ide, Kazuma Ohtaka, Hideko Nagasawa, Yoshihiro Uto, Hitoshi Hori and Kohji Miyazaki : Hypoxia accelerates cancer invasion of hepatoma cells by upregulating MMP expression in an HIF-1alpha-independent manner, International Journal of Oncology, Vol.29, No.6, 1533-1539, 2006.
(要約)
Tumor hypoxia has been reported to induce tumor progression in several carcinomas. Current studies have shown that hypoxia inducible factor-1alpha (HIF-1alpha) is stabilized under hypoxic conditions and transactivates various genes related to cancer aggressiveness. In the present study, we examined whether hypoxia affects cancer invasion in hepatocellular carcinoma. We aimed to solve the molecular mechanism of tumor invasion under the hypoxic condition. We showed that tumor hypoxia accelerated cancer invasion in two hepatoma cell lines. Using Western blot and RT-PCR analyses we demonstrated striking evidence that the expression of HIF-1alpha, ETS-1, MMP-7 and MT1-MMP was strongly upregulated by hypoxic stimulation. To examine whether these invasion-related genes are regulated by HIF-1alpha, we treated hepatoma cells with TX-402, which was reported to repress HIF-1alpha expression. HIF-1alpha expression was strongly repressed by the TX-402 treatment. In contrast, the expression of ETS-1, MMP-7 and MT1-MMP mRNA was not affected by TX-402 treatment. We further established stable transfectants in which HIF-1alpha dominant negative vector was introduced into Hep3B cells (pHIF-1alphaDN). In the pHIF-1alphaDN cells, the expression of ETS-1, MMP-7 and MT1-MMP was not repressed. Moreover, the invasion activity of pHIF-1alphaDN was not altered, compared with that of the mock. In hepatoma cells, we provided evidence that hypoxic stress accelerates cancer invasion by upregulating ETS-1 and the MMP family by an HIF-1alpha-independent pathway.
Kazuto Ohkura, Hitoshi Hori and Hideaki Nagamune : Molecular dynamics of human-specific cytolysin: analysis of membrane binding motif for therapeutic application, Anticancer Research, Vol.26, No.6A, 4055-4062, 2006.
(要約)
Intermedilysin (ILY) is a human-specific cytolysin secreted from Streptococcus intermedius. In this study, the dynamic structure of ILY, StreptolysinO (SLO) and their 12mer substituted mutants for 500 ps was analyzed. Several parameters, such as dipole moment and electrostatic potential, were determined to elucidate the molecular mechanism of membrane binding. Molecular models of lLY, SLO and their mutants were constructed using Insightll-Discover with the Homology module. Their molecular dynamics were simulated with the Discover3 (Insight module), and z-matrix data of the membrane-binding 12mer region were extracted to calculate the molecular orbital (MO) parameters (i.e., dipole moment, solvation free energy (dGW)). Cytolysins vibrated like a bow, and the dipole moment direction of ILY 12mer region was different from that of SLO. Certain ILY mutants indicated the SLO-like dipole properties, which had an SLO-type limer cysteine motif amino acid sequence. The ILY 11mer region was more hydrophobic than that of SLO, and seemed to interact easily with the cell membrane without cholesterol. The electrostatic potential field distribution of ILY differed from that of SLO, especially in the 11mer region. In the 11mer region, the dipole moment directions of these cytolysins were constant during molecular movement, and ready to interact with membrane components (i.e., cholesterol, phospholipid).
Yoshihiro Uto, Shutaro Ae, Daisuke Koyama, Mitsutoshi Sakakibara, Naoki Otomo, Mamoru Otsuki, Hideko Nagasawa, Kenneth L. Kirk and Hitoshi Hori : Artepillin C isoprenomics: design and synthesis of artepillin C isoprene analogues as lipid peroxidation inhibitor having low mitochondrial toxicity, Bioorganic & Medicinal Chemistry, Vol.14, No.16, 5721-5728, 2006.
(要約)
We designed and synthesized isoprene analogues of artepillin C, a major component of Brazilian propolis, and investigated the inhibitory activity on lipid peroxidation of rat liver mitochondria (RLM) and RLM toxicity based on isoprenomics. We succeeded in the synthesis of artepillin C isoprene analogues using regioselective prenylation within the range from 22% to 53% total yield. Reactivity of artepillin C and its isoprene analogues with ABTS (2,2'-Azinobis(3-ethylbenzothiazoline-6-sulfonate)) radical cations showed only a slight difference among the molecules. The isoprene side-chain elongation analogues of artepillin C showed almost the same inhibitory activity against RLM lipid peroxidation as artepillin C. Artepillin C and its isoprene analogues had very weak RLM uncoupling activity. Moreover, artepillin C and its isoprene analogues exhibited a lower inhibitory activity against adenosine 5'-triphosphate (ATP) synthesis by about two orders of magnitude than the effective inhibitory activity against RLM lipid peroxidation. From these results we conclude that artepillin C isoprene analogues could be potent lipid peroxidation inhibitors having low mitochondrial toxicity. We also conclude that elongation of the isoprene side chain of artepillin C to increase lipophilicity had little influence on the inhibitory activity toward RLM lipid peroxidation.
Yoshihiro Uto, Shutaro Ae, Azusa Hotta, Junji Terao, Hideko Nagasawa and Hitoshi Hori : Artepillin C isoprenomics: design and synthesis of artepillin C analogues as antiatherogenic antioxidants, Advances in Experimental Medicine and Biology, Vol.578, 113-118, 2006.
Aya Yoshida, M. Matumoto, H. Hashizume, Y. Oba, T. Tomishige, H. Inagawa, C. Kohchi, M. Hino, F. Ito, K. Tomoda, T. Nakajima, K. Makino, Hiroshi Terada, Hitoshi Hori and Genichiro Soma : Selective delivery of rifampicin incorporated into poly(DL-lactic-co-glycolic) acid microspheres after phagocytotic uptake by alveolar macrophages, and the killing effect against intracellular Mycobacterium bovis Calmette-Guerin, Microbes and Infection, Vol.8, No.9-10, 2484-2491, 2006.
(要約)
Macrophages and their phagocytotic abilities play a dominant role for defense against infected organisms. However, Mycobacterium tuberculosis can survive in the phagosomes of macrophages. In this study, the effective delivery of a drug and the killing effect of tubercle bacilli within macrophages were investigated utilizing the phagocytotic uptake of rifampicin (RFP) that had been incorporated into poly(DL-lactic-co-glycolic) acid (PLGA) microspheres. The microspheres were composed of PLGA that had a monomer ratio (lactic acid/glycolic acid) of either 50/50 or 75/25. They had molecular weights from 5000 to 20,000, and diameters of 1.5, 3.5, 6.2 and 8.9 microm. The most significant factor for phagocytotic activity of macrophages was the diameter of the microspheres. By contrast, molecular weight and monomer ratio of PLGA did not influence phagocytosis. The amount of RFP delivered into cells was also investigated. RFP-PLGA microspheres composed of PLGA with a molecular weight of 20,000 and monomer ratio of 75/25 showed the highest amount of delivery (4 microg/1 x 10(6) cells). Fourteen days after infection, the survival rate of treated intracellular bacilli was 1% when compared with untreated cells. There was almost no killing effect of free RFP (4 or 15 microg/ml) on intracellular bacilli. In vivo efficacy of RFP-PLGA was also examined in rats infected with M. tuberculosis Kurono. Intratracheal administration of RFP-PLGA microspheres was shown to be superior to free RFP for killing of intracellular bacilli and preventing granuloma formation in some lobes. These results suggest that phagocytotic activity could be part of a new drug delivery system that selectively targeted macrophages.
Shin-ichiro Masunaga, Hideko Nagasawa, Yoshinori Sakurai, Yoshihiro Uto, Hitoshi Hori, Kenji Nagata, Minoru Suzuki, Akira Maruhashi, Yuko Kinashi and Koji Ono : The usefulness of mild temperature hyperthermia combined with a newly developed hypoxia-oriented 10B conjugate compound, TX-2100, for boron neutron capture therapy, International Journal of Hyperthermia, Vol.22, No.4, 287-299, 2006.
(要約)
To evaluate the usefulness of a new 10B-compound (TX-2100) as a 10B-carrier in boron neutron capture therapy (BNCT), compared with the simultaneous use of its component drugs, sodium borocapate-10B (BSH) and 3-amino-2-quinoxalinecarbonitrile 1,4-dioxide (TX-402). Further, the usefulness of mild temperature hyperthermia (MTH, 40 degrees Celsius, 30 min) combined with TX-2100 was also examined compared with MTH combined with the concurrent administration with its component drugs. TX-2100 is a hybrid compound that has both a hypoxic cytotoxin unit (TX-402) and a thermal neutron-sensitizing unit (BSH). TX-2100 or both TX-402 plus BSH in combination with MTH or not was administered to SCC VII tumour-bearing mice intra-peritoneally. Then, the 10B concentrations in the tumours and normal tissues were measured by gamma-ray spectrometry. Meanwhile, SCC VII tumour-bearing mice were continuously given 5-bromo-2'-deoxyuridine (BrdU) to label all proliferating (P) cells in the tumours, then treated with TX-2100, TX-402 plus BSH or BSH only, in the same manner as in the biodistribution experiments, either with or without MTH. Right after thermal neutron irradiation during which intra-tumour 10B concentrations remained at similar levels, the tumours were excised, minced and trypsinized. The tumour cell suspensions thus obtained were incubated with cytochalasin-B (a cytokinesis blocker) and the micronucleus (MN) frequency in cells without BrdU labelling (=quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. Meanwhile, the MN frequency in the total (P + Q) tumour cell population was determined from the tumours that were not pre-treated with BrdU. The clonogenic cell survival was also determined in mice given no BrdU. 10B biodistribution analyses in tumours, brain, skin, muscles, blood and liver indicated that the administration of TX-2100 plus MTH is most favourable for concentrating a sufficient amount of 10B in tumours and maintaining a high enough 10B concentration during irradiation. In addition, MTH had a stronger sensitizing effect when combined with TX-2100 than with the concurrent administration of its components TX-402 and BSH on both the total and Q cell populations in solid tumours. MTH was very effective in combination with the newly-developed TX-2100. The sensitizing effect in combination with MTH should be examined when new 10B-carriers are designed.
Shin-ichiro Masunaga, Yoshihiro Uto, Hideko Nagasawa, Hitoshi Hori, Kenji Nagata, Minoru Suzuki, Yuko Kinashi and Koji Ono : Evaluation of Hypoxic Cell Radio-sensitizers in Terms of Radio-sensitizing and Repair-inhibiting Potential Dependency on p53 Status of Tumor Cells and the Effects on Intratumor Quiescent Cells, Anticancer Research, Vol.26, No.2A, 1261-1270, 2006.
(要約)
Intratumor quiescent (Q) cells and p53-mutated tumor cells are more difficult to control than intratumor proliferating (P) cells and p53 wild-type tumor cells, respectively. The usefulness of 3 hypoxic cell radio-sensitizers was compared in terms of a radio-sensitizing effect under aerobic and hypoxic conditions and a repair-inhibiting effect following irradiation on both Q and total (P + Q) cell populations in solid tumors. The dependency of these effects on the p53 status of tumor cells was also examined using tumor cell lines with identical genetic backgrounds except for their p53 status. Human head and neck squamous cell carcinoma cells transfected with mutant TP53 (SAS/mp53) or with neo vector as a control (SAS/neo) were inoculated subcutaneously into both the hind legs of Balb/cA nude mice. The nude mice bearing the tumors and C3H/He mice bearing SCC VII tumors received 5-bromo-2'-deoxyuridine (BrdU) continuously to label all the P cells in the tumors. Tumor-bearing mice received gamma-ray irradiation while alive or following tumor clamping after being administered no drug, nimorazole, SR-2514 or misonidazole, or received no drug, nimorazole, SR-2514 or misonidazole straight after gamma-ray irradiation. For the group irradiated after receiving the drug, the tumors were excised immediately following irradiation, while for the group irradiated before receiving the drug, the tumors were excised 24 h after irradiation. The excised tumors were minced and trypsinized. The tumor cell suspensions thus obtained were incubated with cytochalasin-B (a cytokinesis blocker), and the micronucleus (MN) frequency in the cells without BrdU labelling (= quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. Meanwhile, the MN frequency in the total tumor cell population was determined from the tumors that had not been pretreated with BrdU. The clonogenic cell survival was also determined in the mice given no BrdU. Both the radio-sensitizing effects under aerobic and hypoxic conditions and the repair-inhibiting effects following gamma-ray irradiation increased in the following order: nimorazole < SR-2514 < misonidazole in both total and Q cells in these 3 tumors. Both effects were more marked in the Q cells and p53-mutated tumors than in the total cells and p53-wild tumors, respectively. In terms of controlling radio-resistant Q tumor cells and p53-mutated tumor cells, the combination of radio-sensitizers and conventional radiotherapy is promising both for radio-sensitization and for repair-inhibition, but further study of the toxicity to normal tissues is needed.
Shin-ichiro Masunaga, Hideko Nagasawa, Keiko Gotoh, Yoshinori Sakurai, Yoshihiro Uto, Hitoshi Hori, Kenji Nagata, Minoru Suzuki, Akira Maruhashi, Yuko Kinashi and Koji Ono : Evaluation of Hypoxia-Specific Cytotoxic Bioreductive Agent-Sodium Borocaptate-10B Conjugates, as 10B-Carriers in Boron Neutron,Capture Therary, Radiation Medicine, Vol.24, No.2, 98-107, 2006.
(要約)
To evaluate the usefulness of 5 new 10B-compounds (TX-2091, TX-2095, TX-2097, TX-2100, and TX-2110) as 10B-carriers in boron neutron capture therpy (BNCT). They were conjugates that had been synthesized from a hypoxia-specific cytotoxic bioreductive agent, quinoxaline oxide TX-402, and a clinically used 10B-carrier, sodium borocaptate-10B (BSH). The 5 new compounds were hybrid compounds that have both a hypoxic cytotoxin unit and a thermal neutron-sensitizing unit, BSH. These new compounds and BSH were administered intraperitoneally to SCC VII tumor-bearing mice. Then, the 10B concentrations in the tumors and normal tissues were measured by gamma-ray spectrometry. Subsequently, SCC VII tumor-bearing mice were continuously given 5-bromo-2'-deoxyuridine (BrdU) to label all proliferating (P) cells in the tumors, then treated with TX-2100, which was chosen based on the results of the above-mentioned biodistribution analyses, or BSH in the same manner as in the biodistribution studies. Right after irradiation, during which intratumor 10B concentrations were kept at levels similar to each other, the tumors were excised, minced, and trypsinized. The tumor cell suspensions thus obtained were incubated with cytochalasin-B (a cytokinesis blocker), and the micronucleus (MN) frequency in cells without BrdU labeling [= quiescent (Q) cells] was determined using immunofluorescence staining for BrdU. Meanwhile, the MN frequency in the total (P+Q) tumor cell population was determined from the tumors that were not pretreated with BrdU. Clonogenic cell survival was also determined in mice given no BrdU. 10B biodistribution analyses in tumors, brain, skin, muscles, blood, and liver indicated that TX-2100 has the most favorable characteristics for concentrating a sufficient amount of 10B in tumors and maintaining a high enough 10B concentration during irradiation. In addition, TX-2100 had a significantly stronger radio-sensitizing effect with reactor thermal neutron beams than BSH on both total and Q cells in solid tumors. Further, TX-2100 clearly exhibited a radio-sensitizing effect with gamma-rays not only on total cells but also on Q and hypoxic tumor cells, which was not achieved by BSH. A 10B-carrier that acts as a hypoxic cytotoxin on tumor cells as well as having the potential to keep 10B in tumors and sensitize tumor cells more markedly than conventional 10B-carriers, such as TX-2100, is a promising candidate for use in BNCT.
Mami Hino, Chie Kohchi, Takashi Nishizawa, Aya Yoshida, Kazue Nakata, Hiroyuki Inagawa, Hitoshi Hori, Kimiko Makino, Hiroshi Terada and Genichiro Soma : Innate-immune Therapy for Lung Carcinoma Based on Tissue-macrophage Activation with lipopolysaccharide, Anticancer Research, Vol.25, No.6A, 3747-3754, 2005.
(要約)
Over the last decade, tumor-specific antigens have been discovered, but so far it has not been possible to use them as part of an effective acquired immunotherapy. This failure may be due to the fact that the expression of the MHC class 1 is low and in lung cancer cells is heterogeneous. Therefore, it may be advantageous to develop techniques that activate the antitumor mechanism of the innate immune system. An experimental model was developed for testing lung cancer therapies that are based on the stimulation of macrophages, which then activate innate immunity. A549, a human lung adenocarcinoma cell line, was co-cultured with a rat macrophage cell line (NR8383), or a human macrophage cell line (THP 1) at the ratios of 1:1 or 1:5. The experiments were performed with lipopolysaccharide (LPS) or in its absence. The cytotoxicity rate to A549 cells was estimated over time using a dye-uptake method and the amount of lactate dehydrogenase released was measured. The amount of nitric oxide (NO) induced in the medium was assayed, because it may be a candidate as a useful cytotoxic factor. High cytotoxicity was observed to A549 cells when co-cultured with NR8383 cells in the presence of LPS. This effect was not observed in the absence of LPS. Similar results, although to a lesser extent, were observed when A549 cells were co-cultured with THP-1 cells. A high concentration of NO was measured in the co-culture medium of A549 cells and NR8383 cells when LPS was present. The induction of cell death in lung cancer cells occurred after contact with macrophages that had been activated by LPS. The NO that was produced by macrophages in response to LPS was responsible for some of this effect.
Mami Hino, Mutsumi Oda, Aya Yoshida, Kazue Nakata, Chie Kochi, Takashi Nishizawa, Hiroyuki Inagawa, Hitoshi Hori, Kimiko Makino, Hiroshi Terada and Genichiro Soma : Establishment of an in vitro model using NR8383 cells and mycobacterium bovis calmette-guerin that mimics a chronic infection of mycobacterium tuberculosis., In Vivo, Vol.19, No.5, 821-830, 2005.
(要約)
Mycobacterium tuberculosis infection affects one-third of the world's population and causes the death of three million people each year. To clarify details of M. tuberculosis survival strategies, it is important to establish a suitable in vitro model that mimics a chronic infection in alveolar macrophages by M. tuberculosis. For this reason, we established a new in vitro model using a rat alveolar macrophage cell line, NR8383. Basic characteristics, including phagocytotic ability and production of nitrogen oxide and tumor necrosis factor in response to several stimuli, of NR8383 cells were compared with those of primary alveolar macrophages. The course after phagocytosis of live or killed M. bovis bacilli Calmette-Guerin (BCG) was examined over 21 days using NR8383 cells as the host. The characteristics that have been examined to date were nearly the same for both primary alveolar macrophage and NR8383 cells. Live BCG phagocytosed by NR8383 cells had successfully begun to grow in the cells within 7 days, while killed BCG were almost completely destroyed by 21 days. BCG-infected NR8383 cells are potentially a suitable in vitro model that mimics a chronic infection with M tuberculosis.
Yoshihiro Uto, Shutaro Ae, Hideko Nagasawa and Hitoshi Hori : Artepillin C Isoprenomics: Facile Total Synthesis and Discovery of Amphiphilic Antioxidant, ACS symposium series, No.909, 176-187, 2005.
Shin-ichiro Masunaga, Hideko Nagasawa, Yoshihiro Uto, Hitoshi Hori, K. Ohnishi, A. Takahashi, T. Ohnishi, M. Suzuki, K. Nagata, Y. Kinashi and K. Ono : Combination of the antivascular agent ZD6126 with hypoxic cytotoxin treatment, with reference to the effect on quiescent tumor cells and the dependency on p53 status of tumor cells, Oncology Reports, Vol.14, No.2, 394-400, 2005.
(要約)
Human head and neck squamous cell carcinoma cells transfected with mutant TP53 (SAS/mp53) or with neo vector as a control (SAS/neo) were inoculated subcutaneously into both the hind legs of Balb/cA nude mice. Mice bearing the tumors received 5-bromo-2'-deoxyuridine (BrdU) continuously to label all proliferating (P) cells in the tumors. The mice then received a hypoxic cytotoxin, tirapazamine (TPZ) or TX-402, with or without a vascular targeting agent (VTA), ZD6126. Another group of mice given ZD6126 received a series of test doses of gamma-rays while alive or after tumor clamping to obtain hypoxic fractions (HFs) in the tumors. After each treatment, the tumor cells were isolated and incubated with a cytokinesis blocker (cytochalasin-B), and the micronucleus (MN) frequency in cells without BrdU labeling [quiescent (Q) cells] was determined using immunofluorescence staining for BrdU. The MN frequency in total (P+Q) tumor cells was determined from the tumors that were not pretreated with BrdU. Both hypoxic cytotoxins showed significantly greater toxicity toward SAS/mp53 and Q than SAS/neo and total tumor cells, respectively. The sensitivity to TX-402 was significantly higher than that to TPZ in both total and Q tumor cells of both tumors. The significant enhancive effect by ZD6126 combined with each hypoxic cytotoxin was similar irrespective of p53 status, and slightly greater for total than Q cells probably because of a more marked increase in the size of the HFs in total than Q cells on the use of ZD6126 in both tumors, resulting in a reduction of the difference in the sensitivity to the hypoxic cytotoxin between total and Q cells. In the treatment of conventional cancer therapy-resistant Q tumor cells or p53-mutated tumor cells, the use of hypoxic cytotoxin was very promising either alone or when VTA was co-administered. TX-402 might be more promising than TPZ, although further study of the toxicity to normal tissue is needed.
Shin-ichiro Masunaga, Hideko Nagasawa, Yoshihiro Uto, Hitoshi Hori, Minoru Suzuki, Kenji Nagata, Yuko Kinashi and Koji Ono : The usefulness of continuous administration of hypoxic cytotoxin combined with mild temperature hyperthermia, with reference to effects on quiescent tumor cell populations., International Journal of Hyperthermia, Vol.21, No.4, 305-318, 2005.
(要約)
To evaluate the usefulness of continuous administration of hypoxic cytotoxins in terms of targeting acute hypoxia in solid tumours and the significance of combination with mild temperature hyperthermia (MTH) (40 degrees C, 60 min), the cytotoxic effects of singly or continuously administered tirapazamine (TPZ) and TX-402 were examined in combination with or without MTH in vivo. Further, the effects were also analysed on total (=proliferating (P)+quiescent (Q)) and Q cell populations in solid tumours with the method for selectively detecting the Q cell response. C3H/He mice bearing SCC VII tumours received a continuous administration of 5-bromo-2'-deoxyuridine (BrdU) for 5 days to label all P cells. The tumour-bearing mice then received a single intra-peritoneal injection or 24 h continuous subcutaneous infusion of hypoxic cytotoxin, TPZ or TX-402, with or without MTH. On the other hand, to detect the changes in the hypoxic fraction (HF) in the tumours by MTH, another group of mice with or without MTH received a series of test doses of gamma-rays while alive or after tumour clamping. After each treatment, the tumour cells were isolated and incubated with a cytokinesis blocker (=cytochalasin-B) and the micronucleus (MN) frequency in cells without BrdU labelling (=Q cells) was determined using immunofluorescence staining for BrdU. The MN frequency in total tumour cells was determined from the tumours that were not pre-treated with BrdU. The sensitivity to TX-402 was slightly higher than that to TPZ in both total and Q tumour cells. Continuous administration elevated the sensitivity of both total and Q cells, especially total cells. MTH raised the sensitivity of Q cells more remarkably than that of total cells in both single and continuous administrations. It was thought to be probably because of the higher dose distribution of hypoxic cytotoxin in intermediately hypoxic areas derived mainly from chronic hypoxia through MTH. From the viewpoint of tumour control as a whole including both total and Q tumour cells, the continuous administration of hypoxic cytotoxin combined with MTH may be useful for sensitizing tumour cells in vivo.
C Kohchi, H Inagawa, M Hino, M Oda, K Nakata, Aya Yoshida, Hitoshi Hori, H Terada, K Makino, K Takiguchi and Genichiro Soma : Utilization of macrophages in anticancer therapy: the macrophage network theory, Anticancer Research, Vol.24, No.5C, 3311-3320, 2004.
(要約)
Appropriate and rational modulation of innate immunity may enhance the therapeutic efficacy of emerging immune therapies for treating cancer. One of the crucial cells of innate immunity is the macrophage. The purpose of this article was to review those issues that suggest ways of exploiting macrophage local functions in immune therapy, and to discuss the suitability of low molecular-weight lipopolysaccharides as potent modulators of macrophage functions for immune therapy of cancer.
Hideko Nagasawa, Hideyuki Sasaki, Yoshihiro Uto, Shin-ichi Kubo and Hitoshi Hori : Association of the Macrophage Activating Factor (MAF) Precursor Activity with Polymorphism in Vitamin D-binding Protein, Anticancer Research, Vol.24, No.5c, 3361-3366, 2004.
Shin-ichiro Masunaga, Hideko Nagasawa, Masamitsu Hiraoka, Yoshinori Sakurai, Yoshihiro Uto, Hitoshi Hori, Kenji Nagata, Minoru Suzuki, Akira Maruhashi, Yuko Kinashi and Koji Ono : Applicability of the 2-Nitroimidazole-sodium borocaptate-10B Conjugate, TX-2060, as a 10B-carrier in Boron Neutron Capture Therapy, Anticancer Research, Vol.24, No.5, 2975-2983, 2004.
Hitoshi Hori, Hideko Nagasawa, Yoshihiro Uto, Kazuto Ohkura, Kenneth L. Kirk, Yoshimasa Uehara and Mariko Shimamura : Design of Hypoxia-Trageting Protein Tyrosine Kinase Inhibitor using an innovative pharmacophore 2-methylene-4-cyclopentene-1,3-dione, Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, Vol.1697, No.1-2, 29-38, 2004.
Hisashi Ueta, Hideko Nagasawa, Yuriko Oyabu-Manabe, Kazunori Toida, Kazunori Ishimura and Hitoshi Hori : Localization of enolase in synaptic plasma membrane as an alpha gamma heterodimer in rat brain., Neuroscience Research, Vol.48, No.4, 379-386, 2004.
Y. Nakagawa, S. Shirivichayakul, P. Phanuphak, T. Suda, K. Mito and Hitoshi Hori : Beneficial effect of macrophage activating agent NK-4 on Thai HIV-infected patients, Anticancer Research, Vol.23, No.6A, 4389-4394, 2003.
(要約)
We examined the effect of NK-4 (4,4'-[3-[2-(1-ethyl-4(1H)-quinolinylidene) ethylidene] propenylene] bis(-ethylquinolinium iodide)) on the clinical course of Thai HIV-infected patients receiving antiretroviral drugs. Twelve subjects with CD4 count < 500/mm3 were enrolled to receive 0.5 mg of NK-4 orally once daily for 8 weeks and then twice daily during the follow-up. Most patients showed vitality, increased appetite, stable body-weight, increased CD4 counts and no serious adverse effects due to viral load. We suggest that the increased CD4 counts are due to the macrophage activating effect of NK-4. The patients with increased CD4 counts showed lower alpha-N-acetylgalactosaminidase (alpha-NaGalase) activity than those with decreased CD4 counts. These results suggest that the macrophage activating agent NK-4, as an inexpensive and safe drug for HIV-infected patients, may play a beneficial role in the clinical treatment of HIV-infected patients.
Saharuddin B. Mohamad, Hitoshi Hori, Hideko Nagasawa, Kenji Usui and Yoshihiro Uto : Characterization of human Gc protein-derived macrophage activation factor (GcMAF) and its functional role in macrophage tumoricidal activity, Advances in Experimental Medicine and Biology, Vol.510, 77-82, 2003.
Mariko Shimamura, Hideko Nagasawa, H Ashino, Y Yamamoto, T Hazato, Yoshihiro Uto, Hitoshi Hori and Seiichi Inayama : A novel hypoxia-dependent 2-nitroimidazole KIN-841 inhibits tumour-specific angiogenesis by blocking production of angiogenic factors, British Journal of Cancer, Vol.88, No.2, 307-313, 2003.
Shin-ichiro Masunaga, Koji Ono, Mitsunori Kirihata, Masao Takagaki, Yoshinori Sakurai, Yuko Kinashi, Tooru Kobayashi, Minoru Suzuki, Kenji Nagata, Hideko Nagasawa, Yoshihiro Uto and Hitoshi Hori : Potential of α-amino alcohol p-boronophenylalaninol as a boron carrier in boron neutron capture therapy, regarding its enantiomers, Journal of Cancer Research and Clinical Oncology, Vol.129, No.1, 21-28, 2003.
(要約)
We evaluated the potential of a newly developed (10)B-containing alpha-amino alcohol of p-boronophenylalanine-(10)B (BPA), p-boronophenylalaninol (BPAol), as a boron carrier in boron neutron capture therapy. C57BL mice bearing EL4 tumors received 5-bromo-2'-deoxyuridine (BrdU) continuously via implanted mini-osmotic pumps to label all proliferating (P) cells. After oral administration of L-BPA or D-BPA, or intraperitoneal injection of L-BPAol or D-BPAol, the tumors were irradiated with reactor thermal neutron beams. Some of the tumors were heated at 40 degrees C for 30 min (mild temperature hyperthermia (MTH)) right before neutron exposure, and/or tirapazamine (TPZ) was intraperitoneally injected 30 min before irradiation. The tumors were then excised, minced, and trypsinized. The tumor cell suspensions thus obtained were incubated with cytochalasin-B (a cytokinesis blocker), and the micronucleus (MN) frequency in cells without BrdU labeling [ =quiescent (Q) cells] was determined using immunofluorescence staining for BrdU. Meanwhile, 6 h after irradiation, tumor cell suspensions obtained in the same manner were used for determining the apoptosis frequency in Q cells. The apoptosis and MN frequency in total (P+Q) tumor cells were determined from the tumors that were not pretreated with BrdU. Without TPZ or MTH, L- and D-BPAol increased both frequencies markedly, especially for total cells. Although not significantly larger, L-BPA and D-BPAol increased both frequencies slightly more than D-BPA and L-BPAol, respectively. Combination with both MTH and TPZ markedly reduced the sensitivity difference between total and Q cells. Both L- and D-BPAol have potential as a (10)B-carrier in neutron capture therapy, especially when combined with both MTH and TPZ.
Shin-ichiro Masunaga, Koji Ono, Akihisa Takahashi, Ken Onishi, Takeo Onishi, Minoru Suzuki, Kenji Nagata, Yuko Kinashi, Hideko Nagasawa, Yoshihiro Uto and Hitoshi Hori : Usefulness of combined treatment with mild temperature hyperthermia and/or tirapazamine in the treatment of solid tumors: its independence of p53 states, Cancer Science, Vol.94, No.1, 125-133, 2003.
Saharuddin B. Mohamad, Hideko Nagasawa, Yoshihiro Uto and Hitoshi Hori : Preparation of Gc Protein-derived macrophage Activation Factor (GcMAF) and its Structural Characterization and Biological Activities., Anticancer Research, Vol.22, No.6C, 4297-4300, 2002.
Yoshihiro Uto, Akihiko Hirata, Tomoya Fujita, Syunsuke Takubo, Hideko Nagasawa and Hitoshi Hori : First Total Synthesis of Artepillin C Established by o,o'-Diprenylation of p-Halophenols in Water, The Journal of Organic Chemistry, Vol.67, No.7, 2355-2357, 2002.
Saharuddin B. Mohamad, Hideko Nagasawa, Yoshihiro Uto and Hitoshi Hori : Tumor cell alpha-N-acetylgalactosaminidase activity and its involvement in GcMAF-related macrophage activation, Comparative Biochemistry and Physiology. Part A: Molecular & Integrative Physiology, Vol.132, No.1, 1-8, 2002.
Yoshihiro Uto, Akihiko Hirata, Masaki Ishibashi, Hideko Nagasawa and Hitoshi Hori : Design and synthesis of 2,6-diprenyl-4-iodophenol TX-1952 with a novel and potent anti-peroxidative activity, Comparative Biochemistry and Physiology. Part A: Molecular & Integrative Physiology, Vol.132, No.1, 41-45, 2002.
Chika Nakamura, Nobuhide Kawasaki, Hideki Miyataka, Ezhuthachan Jayachandran, Ho In Kim, Kenneth L. Kirk, Takeo Taguchi, Yoshio Takeuchi, Hitoshi Hori and Toshio Satoh : Synthesis and Biological Activities of Fluorinated Chalcone Derivatives, Bioorganic & Medicinal Chemistry, Vol.10, No.3, 699-706, 2002.
(要約)
We have designed and synthesized new 5-lipoxygenase inhibitors, fluorinated 3,4-dihydroxychalcones, and evaluated their biological activities with respect to antiperoxidation activity and in vitro antitumor activities. All fluorinated chalcones tested showed 5-lipoxygenase inhibition on rat basophilic leukemia-1 (RBL-1) cells and inhibitory action on Fe(3+)-ADP induced NADPH-dependent lipid peroxidation in rat liver microsomes. The potencies were comparable or better to that of the lead 3,4-dihydroxychalcone. 6-Fluoro-3,4-dihydroxy-2',4'-dimethoxy chalcone (7) was the most effective compound in the in vitro assay using a human cancer cell line panel (HCC panel) consisting of 39 systems.
Kazuto Ohkura and Hitoshi Hori : Modification of Cell Response to Insulin by Membrane-acting Agents in Rat White Adipoctes: Analysis of Structural Features by Compytative Simulation, Bioorganic & Medicinal Chemistry, Vol.9, No.11, 3023-3033, 2001.
(要約)
The effect of membrane-acting agents, biscoclaurine alkaloids (cepharanthine, tetrandrine, isotetrandrine), carbobenzoxy-D-Phe-L-Phe-Gly (z-FFG), and tyrphostin AG17, on the insulin-involved fatty acid synthesis by an beta-agonist (e.g., isoproterenol) in adipocytes was examined. The alkaloids dose-dependently enhanced the insulin-involved fatty acid synthesis in rat white adipocytes, stabilized the C(6)-NBD-PC (1-acyl-2-[6-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-caproyl]-sn-glycero-3-phosphatidylcholine) model membrane, and suppressed the phospholipase A(2)-induced phospholipid degradation. In contrast, z-FFG had no effect on the fatty acid synthesis or the membrane stability. Tyrphostin AG17 suppressed insulin action, but promoted the model membrane stabilization. In the same culture conditions as for the fatty acid synthesis assay, cepharanthine, z-FFG and tyrphostin AG17 had no effect on the transcript levels of glucose transporter isoforms (GLUT 1, 4) and hexokinase isozymes (HK I, II) in rat white adipocytes. Thus, these membrane-acting agents modify the insulin action via a change in the cell membrane condition, and do not directly act on the insulin-involved glucose metabolism. Then we analyzed the structural conformation of these membrane-acting agents by computational simulations. The alkaloids had an elliptic macrocyclic structure, and the order of ellipticity (cepharanthine>tetrandrine>isotetrandrine) agreed with that of the modifying ability for insulin action. The distribution of electrostatic potential fields of these alkaloids was essentially equal by turn in surrounding with the dipole moments. Both in z-FFG and tyrphostin AG17, the distribution pattern of electrostatic potential fields was different from that of the alkaloids. Judging from these results, we concluded that the electrostatic potential field is a good index of the modification of insulin action, and the elliptic structure in these alkaloids is regarded with the modification of insulin action.
Shin-ichi Uesato, Yutaka Kitagawa, Masaaki Kamishimoto, Ayako Kumagai, Hitoshi Hori and Hideko Nagasawa : Inhibition of green tea catechins against the growth of cancerous human colon and heptatic epithelial cells, Cancer Letters, Vol.170, No.1, 41-44, 2001.
Shin-ichiro Masunaga, Koji Ono, Mitsunori Kirihata, Masao Takagaki, Yoshinori Sakurai, Yuko Kinashi, Tooru Kobayashi, Hideko Nagasawa, Yoshihiro Uto and Hitoshi Hori : EVALUATION OF THE POTENTIAL OF PARA-BORONOPHENYLALANINOL AS A BORON CARRIER IN BORON NEUTRON CAPTURE THERAPY, REFERRING TO THE EFFECT ON INTRATUMOR QUIESCENT CELLS, Japanese Journal of Cancer Research, Vol.92, No.9, 996-1007, 2001.
Shin-ichiro Masunaga, Koji Ono, Minoru Suzuki, Yasumasa Nishimura, Yuko Kinashi, Masao Takagaki, Hitoshi Hori, Hideko Nagasawa, Yoshihiro Uto, Izumi Tsuchiya, Sotaro Sadahiro and Chieko Murayama : Radiosensitization effect by combination with paclitaxel in vivo, including the effect on intratumor quiscent cells, International Journal of Radiation Oncology*Biology*Physics, Vol.50, No.4, 1063-1072, 2001.
Shin-ichiro Masunaga, Koji Ono and Hitoshi Hori : Exploiting tumor hypoxia in the treatment of solid tumors, Japanese Journal of Hyperthermic Oncology, Vol.17, No.1, 13-22, 2001.
111.
Shin-ichiro Masunaga, Koji Ono, Hitoshi Hori, Minoru Suzuki, Yuko Kinashi, Masao Takagaki, Soko Kasai, Hideko Nagasawa and Yoshihiro Uto : Change in oxygenation status in intratumour total and quienscent cells following γ-ray irradiation,tirapazamine administration, cisplatin injection and bleomycin treatment, The British Journal of Radiology, Vol.73, No.873, 978-986, 2000.
Shin-ichiro Masunaga, Koji Ono, Minoru Suzuki, Yuko Kinashi, Masao Takagaki, Soko Kasai, Hideko Nagasawa, Yoshihiro Uto and Hitoshi Hori : Changes in the Sensitivity of Intratumor Cells during Fractionated Tirapazamine Administration, Japanese Journal of Cancer Research, Vol.91, No.7, 731-736, 2000.
Shin-ichiro Masunaga, Koji Ono, Minoru Suzuki, Yuko Kinashi, Masao Takagaki, Hitoshi Hori, Soko Kasai, Hideko Nagasawa and Yoshihiro Uto : Usefulness of Tirapazamine as a Combined Agent in Chemoradiation and Thermo-chemoradiation Therapy at Mild Temperatures: Reference to the Effect on Intratumor Quiescent Cells, Japanese Journal of Cancer Research, Vol.91, No.5, 566-572, 2000.
Ji-Wen Zhu, Hideko Nagasawa, Fumi Nagura, Saharuddin B. Mohamad, Yoshihiro Uto, Kazuto Ohkura and Hitoshi Hori : Elucidation of Strict Structural Requirements of Brefeldin A as an Inducer of Differentiation and Apoptosis, Bioorganic & Medicinal Chemistry, Vol.8, No.2, 455-463, 2000.
Hideko Nagasawa, Fumi Nagura, Saharuddin B. Mohamad, Ji-Wen Zhu, Takahiko Tsukuda, T. Hashimoto, Yoshimori Asakawa and Hitoshi Hori : Apoptosis induction in HCT116 cells by cytochalasins isolated from the fungus Daldinia vernicosa, Phytomedicine, Vol.6, No.6, 403-409, 2000.
Kazuto Ohkura and Hitoshi Hori : Analyses of Insulin-Potentiating Fragments of Human Growth Hormone by Computative Simulation; Essential Unit for Insulin-Involved Biological Responses, Bioorganic & Medicinal Chemistry, Vol.8, No.7, 1733-1740, 2000.
(要約)
We analyzed the structural features of insulin-potentiating fragments of human growth hormone by computative simulations. The peptides were designated from the N-terminus sequences of the hormone positions at 1-15 (hGH(1-15); H2N-Phe1-Pro2-Thr3-Ile4-Pro5-Leu6-Ser7-Arg8-L eu9-Phe10-Asp11-Asn12-Ala13-Met14-Leu15 -COOH), 6-13 (hGH(6-13)), 7-13 (hGH(7-13)) and 8-13 (hGH(8-13)), which enhanced insulin-producing hypoglycemia. In these peptide molecules, ionic bonds were predicted to form between 8th-arginyl residue and 11th-aspartic residue, and this intramolecular interaction caused the formation of a macrocyclic structure containing a tetrapeptide Arg8-Leu9-Phe10-Asp11. The peptide positions at 6-10 (hGH(6-10)), 9-13 (hGH(9-13)) and 10-13 (hGH(10-13)) did not lead to a macrocyclic formation in the molecules, and had no effect on the insulin action. Although beta-Ala13hGH(1-15), in which the 13th-alanine was replaced by a beta-alanyl residue, had no effect on insulin-producing hypoglycemia, the macrocyclic region (Arg8-Leu9-Phe10-Asp11) was observed by the computative simulation. An isothermal vibration analysis of both of beta-Ala13hGH(1-15) and hGH(1-15) peptide suggested that beta-Ala13hGH(1-15) is molecule was more flexible than hGH(1-15); C-terminal carboxyl group of Leu15 easily accessed to Arg8 and inhibited the ionic bond formation between Arg8 and Asp11 in beta-Ala13hGH(1-15). The peptide of hGH(8-13) dose-dependently enhanced the insulin-involved fatty acid synthesis in rat white adipocytes, and stabilized the C6-NBD-PC (1-acyl-2-[6-[(7-nitro-2,1,3benzoxadiazol-4-yl)amino]-caproyl]-sn- glycero-3-phosphatidylcholine) model membranes. In contrast, hGH(9-13) had no effect both on the fatty acid synthesis and the membrane stability. In the same culture conditions as the fatty acid synthesis assay, hGH(8-13) had no effect on the transcript levels of glucose transporter isoforms (GLUT 1, 4) and hexokinase isozymes (HK I, II) in rat white adipocytes. Judging from these results we considered that the macrocyclic structure in human growth hormonal peptides is regarded with the modification of insulin action, and hGH(8-13) is an essential sequence for the modification of insulin action. This hGH(8-13) peptide modifies the insulin action via stabilizing the cell membrane, and does not directly act on the insulin-involved glucose metabolism.
Taisuke Inomata, Yasuhiro Ogawa, Akihito Nishioka, Norihiko Hamada, Satoshi Ito, Shinnji Kariya, Shoji Yoshida, Hideko Nagasawa, Hitoshi Hori and Seiichi Inayama : The immunopotentiation effects of the bifunctional radiosensitizer KIN-806 in comparison with its analogs KIN-804 and KIN-844, Oncology Reports, Vol.6, No.6, 1209-1212, 1999.
Shin-ichiro Masunaga, Koji Ono, Hitoshi Hori, Minoru Suzuki, Yuko Kinashi, Masao Takagaki, Soko Kasai, Hideko Nagasawa and Yoshihiro Uto : Potentially Lethal Damage Repair by Total and Quiescent Tumor Cells Following Various DNA-damaging Treatments, Radiation Medicine, Vol.17, No.4, 259-264, 1999.
Chengzhe Jin, Zheshan Quan and Hitoshi Hori : The Design and Synthesis of 2-nitroimidazoles Compounds as a Hypoxic Cell Radiosensitizer and Their Anti-angiogenic Activity, Chinese Journal of Medicinal Chemisty, Vol.9, No.1, 21-26, 1999.
123.
Shin-ichiro Masunaga, Koji Ono, Hitoshi Hori, Yuko Kinashi, Minoru Takagaki, Minoru Suzuki, Soko Kasai, Hideko Nagasawa and Yoshihiro Uto : Modification of tirapazamine-induced cytotoxycity in combination with mild hyperthermia and/or nicotinamide: reference to effect on quiescent tumour cells, Int. J. Hyperthermia, Vol.15, No.1, 7-16, 1999.
Ichiro Shimizu, Yue-Rong Ma, Yoko Mizobuchi, Fei Liu, Tetsuo Miura, Yoichiro Nakai, Mitugi Yasuda, Masako Shiba, Takahiro Horie, Sakae Amagaya, Norifumi Kawada, Hitoshi Hori and Susumu Ito : Effects of Sho-saiko-to, a Japanese Herbal Medicine, on Hepatic Fibrosis in Rats, Hepatology, Vol.29, No.1, 149-160, 1999.
(要約)
It has been shown that lipid peroxidation is associated with hepatic fibrosis and stellate cell activation. Sho-saiko-to (TJ-9) is an herbal medicine, which is commonly used to treat chronic hepatitis in Japan, although the mechanism by which TJ-9 protects against hepatic fibrosis is not known. As a result, we assayed the preventive and therapeutic effects of TJ-9 on experimental hepatic fibrosis, induced in rats by dimethylnitrosamine (DMN) or pig serum (PS), and on rat stellate cells and hepatocytes in primary culture, and assessed the antioxidative activities and the active components of TJ-9. Male Wistar rats were given a single intraperitoneal injection of 40 mg/kg DMN or 0.5 mL PS twice weekly for 10 weeks. In each model, rats were fed a basal diet throughout, or the same diet, which also contained 1.5% TJ-9, for 2 weeks before treatment or for the last 2 weeks of treatment. TJ-9 suppressed the induction of hepatic fibrosis, increased hepatic retinoids, and reduced the hepatic levels of collagen and malondialdehyde (MDA), a production of lipid peroxidation. Immunohistochemical examination showed that TJ-9 reduced the deposition of type I collagen and the number of alpha-smooth muscle actin (alpha-SMA) positive-stellate cells in the liver and inhibited, not only lipid peroxidation in cultured rat hepatocytes that were undergoing oxidative stress, but also the production of type I collagen, alpha-SMA expression, cell proliferation, and oxidative burst in cultured rat stellate cells. In addition, TJ-9 inhibited Fe2+/adenosine 5'-diphosphate-induced lipid peroxidation in rat liver mitochondria in a dose-dependent manner and showed radical scavenging activity. Among the active components of TJ-9, baicalin and baicalein were found to be mainly responsible for the antioxidative activity. These findings suggest that Sho-saiko-to (TJ-9) functions as a potent antifibrosuppressant by inhibition of lipid peroxidation in hepatocytes and stellate cells in vivo.
Kazuto Ohkura and Hitoshi Hori : Analysis of Structure-Permeability Correlation of Nitrophenol Analogues in Newborn Rat Abdominal Skin Using Semiempirical Molocular Orbital Calculation, Bioorganic & Medicinal Chemistry, Vol.7, No.2, 309-314, 1999.
(要約)
Theoretical analysis of the permeation process in percutaneous absorption is important for the molecular design of bioavailable transdermal drugs. In the present study, we examined the difference in permeability of nitrophenols across newborn rat abdominal skin and analyzed the structure-permeability correlation by molecular orbital calculation. The permeable rate of o-, n-, p-nitrophenol and 2,4-, 2,5-, 2,6-dinitrophenols was 0.291, 0.212, 0.085 and 0.042, 0.109, 0.027 micromol/cm2/h, respectively. The permeability of the nitrophenols correlated with their pKa values, indicating the ionizing process related to the permeation through the skin. The pKa values better correlated with the ionization potential (IP) energies than lowest unoccupied molecular orbital (LUMO) energies. Solvation free energies (dGW) of molecular form nitrophenols correlated better with permeabilities (pA) than partition coefficients (log P). In analyzing the dGW values with the permeability at pH 7.4, o-nitrophenol outlay from the theoretical line by ortho-effect. We conclude solvation free energy is a practical parameter and very useful for the molecular design of transdermal drugs.
Hitoshi Hori, Masaki Ishibashi, Saharuddin B. Mohamad, Hideko Nagasawa, Yoshihiro Uto, Hiroyuki Sakamaki, Ning Pan, Kazuto Ohkura and Sansei Nishibe : Enhancement Effect of Methyl α-D-Glucoside for Inhibitory Effects of Antioxidants on ADP/Fe2+-Induced Lipid Peroxidation in Rat Liver Mitochondria, Advances in Experimental Medicine and Biology, Vol.471, 395-401, 1999.
Yoko Mizobuchi, Ichiro Shimizu, Mitugi Yasuda, Hitoshi Hori, Masayuki Shono and Susumu Ito : Retinyl palmitate reduces hepatic fibrosis in rats induced by dimethylnitrosamine or pig serum, Journal of Hepatology, Vol.26, No.6, 933-943, 1998.
Shin-ichiro Masunaga, Koji Ono, Hitoshi Hori, Ken-ichi Akaboshi, Minoru Suzuki, Yuko Kinashi, Soko Kasai, Hideko Nagasawa and Yoshihiro Uto : Enhancement of Cisplatin Sensitivity of Quiescent Cells in Solid Tumors by Combined Treatment with Tirapazamine and Low-Temperature Hyperthermia, Radiation Medicine, Vol.16, No.6, 441-448, 1998.
Shin-ichiro Masunaga, Koji Ono, Yoshinori Sakurai, Hitoshi Hori, Tooru Kobayashi, Masao Takagaki, Minoru Suzuki, Yuko Kinashi and Mitsuhiko Akaboshi : Appplicability of Combination with Tirapazamine in Boron Neutron Capture Therapy, Japanese Journal of Cancer Research, Vol.89, No.7, 768-774, 1998.
(要約)
SCC VII tumor-bearing mice were continuously given 5-bromo-2'-deoxyuridine (BrdU) to label all proliferating cells. After injection of tirapazamine (TPZ), a bioreductive agent, combined with sodium borocaptate-10B (BSH) or dl-p-boronophenylalanine-10B (BPA) administration, the tumors were irradiated with thermal neutrons, and then isolated and incubated with cytochalasin-B (a cytokinesis blocker). The micronucleus (MN) frequency in cells without BrdU labeling (quiescent (Q) cells) was determined by means of immunofluorescence staining for BrdU, and that for total cells was obtained from tumors not pretreated with BrdU. Even when no 10B-compound was administered, TPZ increased the MN frequency of tumor cells including Q cells, resulting in reduction of the difference in MN frequency between total and Q cells, mainly by increasing the MN frequency of Q cells. TPZ increased the MN frequency of Q cells when combined with BPA administration, but TPZ showed no apparent effect on each cell population when combined with BSH. Namely, TPZ reduced the difference in MN frequency between total and Q cells caused by 10B-compound administration, especially when BPA was administered. From the viewpoint of the overall cell killing effect in boron neutron capture therapy (BNCT), combination with TPZ appeared to be useful in BPA-BNCT, but not in BSH-BNCT.
Satoru GOTO, Kai-Xian Chen, Sanae Oda, Hitoshi Hori and Hiroshi Terada : Dynamic Structural Features of Macrocyclic Cytochalasin Analogues Responsible for Their Hexose Transport Inhibition, Journal of the American Chemical Society, Vol.120, No.10, 2457-2463, 1998.
Kazuto Ohkura, Hitoshi Hori and Hiroshi Terada : Effect of Extracellular Calcium on the Intracellular Calcium Level of Newborn Rat Skin Basal Cells, Biological & Pharmaceutical Bulletin, Vol.21, No.1, 1-4, 1998.
(要約)
The effect of extracellular calcium on the intracellular calcium level of newborn rat skin basal cells was examined. The intracellular calcium level of basal cells was affected by the extracellular calcium concentration on culture of the cells on a collagen-coated filter, but not on a plate. The intracellular calcium ion concentration ([Ca2+]i) of cells on a collagen-coated filter with 0.05 mM external CaCl2 was 15 nM and increased to 20-22 nM on culture with 2.0 mM extracellular CaCl2. From these results we concluded that the culture matrix (e.g. culture plates, or collagen-coated filters) affects the response of the basal cells to changes in the extracellular calcium content. The intracellular calcium level of Swiss 3T3 cells cultured both on plates and collagen-coated filters were affected by the extracellular calcium concentration. Their [Ca2+]i was determined as 49-50 nM in the presence of 0.05 mM external CaCl2, and 54-56 nM in the presence of 2.0 mM CaCl2. These different responses to extracellular Ca2+ may be due to differences in the proliferative profiles of basal cells and fibroblasts.
(キーワード)
Animals / Animals, Newborn / カルシウム (calcium) / Culture Media / Extracellular Space / Indicators and Reagents / Mice / Rats / Rats, Wistar / Skin
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 9477160
Shin-ichiro Masunaga, Koji Ono, Hitoshi Hori, Toru Shibata, Minoru Suzuki, Yuko Kinashi, Masao Takagaki and Mitsuhiko Akaboshi : Effects of Bioreductive Agents, Tirapazamine and Mitomycin C, on Quiescent Cell Populations in Solid Tumors, Evaluated by Micronucleus Assay, Japanese Journal of Cancer Research, Vol.88, No.9, 907-914, 1997.
(要約)
Mice bearing transplantable solid tumors received 10 intraperitoneal administrations of 5-bromo-2'-deoxyuridine (BrdU) to label the proliferating (P) tumor cells, and were then irradiated with 60Co gamma-rays or injected with cis-diamminedichloroplatinum (II) (cisplatin). The tumor cells were isolated and incubated with cytochalasin-B (a cytokinesis blocker). The micronucleus (MN) frequency in the cells without BrdU labeling, which were regarded as quiescent (Q) cells in the tumor, was determined using immunofluorescence staining for BrdU. The MN frequency in the total (P + Q) tumor cell population was determined from tumors that were not pretreated with BrdU. Pretreatment with tirapazamine, a bioreductive agent, could enhance the sensitivity of tumor cells, including Q cells, to radiation more markedly than mitomycin C pretreatment as judged from an in vivo assay immediately after irradiation. Post-irradiation administration of tirapazamine produced a large post-irradiation radiosensitizing effect on both the total and Q tumor cell populations in vivo. Cisplatin treatment combined with tirapazamine demonstrated that tirapazamine also has a chemosensitizing potential for both the total and Q tumor cell populations. We confirmed that the sensitivity of Q cell populations to radiation and chemotherapy using cisplatin can be enhanced by combined treatment with tirapazamine.
Tatsuya Fujimoto, Hitoshi Hori, Hideakira Yokoyama, Ning Pan, Miki Kurosaki and Hideko Nagasawa : Respiratory activities of liver mitochondria, isolated from freshwater turtle chinemys revesii as an experimental anoxia-tolerant model system, determined by mitochondrial modifiers, Pathophysiology, Vol.4, No.3, 183-190, 1997.
Ji-Wen Zhu, Hitoshi Hori, Hisao Nojiri, Takahiko Tsukuda and Zenei Taira : Synthesis and Activity of Brefeldin a Analogs as Inducers of Cancer Cell Differentiation and Apotosis, Bioorganic & Medicinal Chemistry Letters, Vol.7, No.2, 139-144, 1997.
137.
Hitoshi Hori, Cheng-Zhe Jin, Masatoshi Kiyono, Soko Kasai, Mariko Shimamura and Seiichi Inayama : Design, Synthesis, and Biological Activity of Anti-angiogenic Hypoxic Cell Radiosensitizer Haloacetylcarbamoyl-2-nitroimidazoles, Bioorganic & Medicinal Chemistry, Vol.5, No.3, 591-599, 1997.
(要約)
We designed, synthesized, and evaluated haloacetylcarbamoyl-2-nitroimidazoles, including chloro (KIN-1800, TX-1835, and TX-1836) and bromo derivatives (TX-1844, TX-1845, and TX-1846), as potential hypoxic cell radiosensitizers with antiangiogenic activities. To establish biological function owing to the haloacetylcarbamoyl group in the side-chain, we compared their in vitro radiosensitizing activities with those of their parent 2-nitroimidazoles without haloacetylcarbamoyl groups: misonidazole (MISO), TX-1831, and TX-1832, respectively. Both tert-butoxy substituted derivatives. TX-1835 and TX-1845, were more potent radiosensitizers than TX-1831. The p-tert-butylphenoxy-substituted derivatives, TX-1836 and TX-1846, and the methoxysubstituted derivatives, KIN-1800 and TX-1844, were stronger radiosensitizers than TX-1832 and MISO. We examined the anti-angiogenic activities of these 2-nitroimidazole derivatives containing haloacetylcarbamoyl group by the rat lung endothelial (RLE) cell proliferation assay and chick embryo chorioallantoic membrane (chick CAM) angiogenesis assay and showed that haloacetylcarbamoyl-2-nitroimidazoles were more potent angiogenic inhibitors than the corresponding desacetylcarbamoyl-2-nitroimidazoles. The in vivo chick CAM angiogenesis assay showed that the strong bromoacetylcarbamoyl-2-nitroimidazole radiosensitizers, such as TX-1845 and TX-1846, were the strongest angiogenic inhibitors among them. We concluded that the bromoacetylcarbamoyl-2-nitroimidazole radiosensitizers, such as TX-1845 and TX-1846, are promising as anti-angiogenic hypoxic cell radiosensitizers.
Eiji Majima, Satoru GOTO, Hitoshi Hori, Yasuo Shinohara, Yeong-Man Hong and Hiroshi Terada : Stablilities of the fluorescent SH-reagent eosin-5-maleimide and its adducts with sulfhydryl compounds, Biochimica et Biophysica Acta (BBA) - General Subjects, Vol.1243, No.3, 336-342, 1995.
(要約)
The stabilities of the SH-reagent eosin-5-maleimide (EMA) and its adducts with the SH-compounds L-cysteine, N-acetyl-L-cysteine and glutathione (reduced form) were studied under various conditions in comparison with those of the adducts of N-ethylmaleimide (NEM). Studies by reversed-phase high performance liquid chromatography and mass spectrometry showed that EMA was less stable than NEM at neutral and moderately alkaline pH values. EMA formed a succinimide-type adduct with SH-compounds, and then underwent further modification by nucleophilic attack of OH- or an amino group. The succinimide-type adducts with acetylcysteine and glutathione were converted to open-type adducts, in which the succinimide ring was cleaved, whereas the adduct with cysteine was modified to a thiazine-type adduct. Kinetic analyses showed that these open-type and thiazine-type adducts were readily formed and were stable at moderately alkaline pH values such as pH 8.0 or 9.0.
Hitoshi Hori, Hideakira Yokoyama, Hideko Nagasawa, Chieko Murayama, Tomoyuki Mori, S Yonei, Tetsuo Sato and Seiichi Inayama : A Convenient Screening Test for Hypoxic Cell Radiosensitizers/Cytotoxins, Biological & Pharmaceutical Bulletin, Vol.17, No.12, 1676-1678, 1994.
Takuhito Tada, Toshibumi Nakajima, Yasuto Onoyama, Chieko Murayama, Tomoyuki Mori, Hideko Nagasawa, Hitoshi Hori and Seiichi Inayama : In Vivo Radiosensitizing Effect of Nitroimidazole Derivative KIN-804, International Journal of Radiation Oncology*Biology*Physics, Vol.29, No.3, 601-605, 1994.
Satoru GOTO, Zong-Ru Guo, Yukako Futatsuishi, Hitoshi Hori, Zenei Taira and Hiroshi Terada : Quantitative structure-activity relationships of benzamide derivatives for anti-leukotriene activities, Journal of Medicinal Chemistry, Vol.35, No.13, 2440-2445, 1992.
(要約)
To determine the structural requirements of the benzamide derivatives reported by Nakai et al. (J. Med. Chem. 1988, 31, 84-91) for antileukotriene activity, we studied their conformational characteristics in comparison with those of leukotriene. By superimpositions of the conformations of antagonists on that of leukotriene, we found that the conformations of the conjugated benzamide moiety, tetrazole ring, and benzopyran or benzodioxan ring of the antagonists correspond to the triene moiety, peptide carboxylic acid residue, and cysteine residue of leukotriene, respectively, but that no moiety of the antagonists corresponds to the terminal aliphatic carboxylic acid moiety of leukotriene. Furthermore, the stable conformations of alkyl and alkoxy groups of the antagonists were quite different from that of the omega-chain of leukotriene. However, conformational analyses taking all the possible rotations of these flexible chains into consideration showed that antagonists in which these flexible chains can most feasibly adopt the same lengths as those of the omega-chain exhibit potent antagonist activity. From these results, we deduced the structural features of benzamide derivatives necessary for potent antileukotriene activity.
Hideko Nagasawa, Masahiro Bando, Hitoshi Hori, Tetsuo Sato, Takuhito Tada, Yasuto Onoyama and Seiichi Inayama : Radiosensitizing, Toxicological, and Pharmacokinetic Properties of Hydroxamate Analogues of Nitroimidazoles as Bifunctional Radiosensitizers/Chemical Modifiers, International Journal of Radiation Oncology*Biology*Physics, Vol.22, No.3, 561-564, 1992.
Hiromu Satake, Hitoshi Hori and Shoji Kaneshina : A Coated Wire Electrode Sensitive to Tetraphenylphosphonium Ion for Measurement of the Mitochondrial Membrane Potential, Analytical Letters, Vol.24, No.2, 295-304, 1991.
Keisuke Sasai, Yuta Shibamoto, Masaharu Takahashi, L. Zhou, Hitoshi Hori, Hideko Nagasawa, Tetsuichi Shibata, Seiichi Inayama and Mitsuyuki Abe : KIH-802, An Ace-tohydroxamic Acid Derivative of 2-Nitroimidazole, as A New Potent Hypoxic Cell Ra-diosensitizer: Radiosensitizing Activity, Acute Toxicity, and Pharmacokinetics, Cancer Chemotherapy and Pharmacology, Vol.26, No.2, 112--116, 1990.
H. Fukuda, T. Tada, M. Tsumura, T. Nakajima, Y. Onoyama, Hitoshi Hori and Seiichi Inayama : Radiosensitizing Effects of Nitroimidazole Derivatives RP170 and KIH-802, J. Osaka City Medical Center, Vol.39, No.3, 573-593, 1990.
147.
Shin-ichi Suzuki, Takako Inoue, Hitoshi Hori and Seiichi Inayama : Analysis of Methamphetamine in Hair, Nail, Sweat, and Saliva by Mass Fragmentography, Journal of Analytical Toxicology, Vol.13, No.3, 176-178, 1989.
(要約)
A method for the detection and quantitation of methamphetamine and its major metabolite in hair, nails, sweat, and saliva from habitual users of methamphetamine by mass fragmentography has been developed. Hair and nail samples were washed with water and methanol to remove the external contamination, processed with 0.6M HCl, alkalinized, and extracted with CHCl3/isopropanol (3:1 v/v). Sweat and saliva samples were extracted with methanol. After trifluoroacetyl derivatization, the samples were analyzed by mass fragmentography. Methamphetamine and its major metabolite, amphetamine, were detected in hair, nail, and sweat samples, but methamphetamine alone was detected in saliva samples.
(キーワード)
Amphetamine / Gas Chromatography-Mass Spectrometry / Hair / Humans / Methamphetamine / Nails / Saliva / Sweat
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 2755090
Hitoshi Hori, Chieko Murayama, Tomoyuki Mori, Yuta Shibamoto, Mitsuyuki Abe, Yasuto Onoyama and Seiichi Inayama : KIH-802: 2-Nitroimidazole-1-acetohydroxamate as a hypoxic Cell Radiosensitizer, International Journal of Radiation Oncology*Biology*Physics, Vol.16, No.4, 1029-1032, 1989.
(要約)
We have identified potassium 2-nitroimidazole-1-acetohydroxamate (KIH-802) as a hypoxic cell radiosensitizer potentially superior to Miso. The water-soluble acetohydroxamates of 2-nitroimidazole (KIH-802; free acid 801) and 4-nitroimidazole (KIH-852) were designed, synthesized, and evaluated by in vitro and in vivo screening against EMT6 cells. Enhancement ratios of KIH-802 and 801 were 1.92 and 1.68, respectively, compared with 1.58 for MISO all at 1 mM. These acetohydroxamates are also expected to be more effective in vitro than SR-2508 based on our previous experiments. In vivo ERs of KIH-802, 801, and 852 were 1.75, 1.50, and 1.35, respectively, compared with 1.57 for MISO all at the same dose of 200 mg/kg. The data clearly show that the addition of an acetohydroxamic acid moiety to the 2-nitroimidazole skeleton can enhance radiosensitizing ability.
G.-M. Pang, C.-J. Zhao, 堀 均, Seiichi Inayama : Studies on New Triterpenoids of Tripterygium regelii, Yao Xue Xue Bao, Vol.24, No.1, 75-79, 1989年.
(要約)
Four triterpenoids were isolated from the root of Tripterygium regelii collected in Fusong Prefecture, Jilin Province. III was confirmed as methyl 3-oxo-22 alpha-acetoxy-23-hydroxy-urs-12-ene-30-oate, named regelin C, V and VI were a pair of isomers identified as methyl 3 beta, 22 alpha-dihydroxy-urs-12-ene-30-oate and methyl 3 beta, 22 alpha-dihydroxy-olean-12-ene-29-oate, named regelindiol A and regelindiol B respectively. VII was represented as methyl 3-oxo-22 alpha-hydroxy-olean-12-ene-29-oate, named regelin D. III, V and VII were new compounds, and VI was isolated for the first time as a natural compound.
(キーワード)
Chemical Phenomena / 化学 (chemistry) / Drugs, Chinese Herbal / Triterpenes
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 2801129
Seiichi Inayama, Hitoshi Hori, G.-M. Pang, Hideko Nagasawa, Yoichi Iitaka and H. Ageta : Isolation of a Hopane-type Trterpenoid, Zeorin, From Higher Plant, Tripterygium regelii, Chemical & Pharmaceutical Bulletin, Vol.37, No.20, 2836-2837, 1989.
151.
Kenzo Harimaya, Hitoshi Hori, Tamiko Ohkura, Takeshi Kawamata, Ji-Fu Gao and Seiichi Inayama : Isolation and Strucuture of New Pseudoquianolides, 4-Epipulchelin and 4-Epineopulchellin From Gaillardia pulchella, Heterocycles, Vol.27, No.1, 83-87, 1988.
152.
H. Koike, Hitoshi Hori, Seiichi Inayama and Hiroshi Terada : Effect of Arylidene-cyclopentenedione Radiosensitizers on ATP Synthesis in Mitochondria:Action as Potent Inhibitors of phosphate Transport, Biochemical and Biophysical Research Communications, Vol.155, No.2, 1066-1074, 1988.
(要約)
The effects of the arylidene-cyclopentenedione radiosensitizers, KIH-200, 201 and 202 on ATP synthesis in mitochondria were examined. In spite of the close similarity of their chemical structure to that of the most potent known weakly acidic uncoupler, SF 6847, they did not show any uncoupling activity at concentrations of up to 50 microM. However, these three compounds were found to have very potent inhibitory effects on Pi-transport into mitochondria, all causing 50% inhibition (I50%) at about 7 microM. Thus they are much more potent than the commonly used Pi-transport inhibitors N-ethylmaleimide (I50% = about 40 microM), and mersalyl (I50% = about 30 microM). They may act as SH-reagents, and inhibit Pi-transport by modifying an SH-group(s) in the Pi-transporter.
M. Tanaka, H. Fukuda, T. Nakajim, K. Sugimoto, Y. Onoyama, Hitoshi Hori and Seiichi Inayama : Radiosensitizing Effect of the Nitroimidazole Derivatives RK28 and DNIE on Murine Tumors, Nippon Acta Radiologica, Vol.48, No.10, 1249-1260, 1988.
154.
K. Takanuki, T. Igarashi, K. Hata, Hitoshi Hori, T. Shibata, H. Kitagawa, T. Satoh and Seiichi Inayama : Glutathione Depletion by 2-Nitroimidazole-1-acetohydroxamic Acid as a Radiosensitizer in Hypoxic Rat Hepatocytes, Biochemistry International, Vol.17, No.1, 155-162, 1988.
155.
Hitoshi Hori, H. Maezawa, Yoichi Iitaka, T. Ohsaka, T. Shibata, T. Mori and Seiichi Inayama : 2-Arylidene-4-cyclopenten-1,3-diones Designed as Non-nitro Radiosensitizers and Hypoxic Cytotoxins, Japanese Journal of Cancer Research, Vol.78, No.10, 1128-1133, 1987.
(要約)
The radiosensitizing effects and hypoxic cytotoxicity of 2-(4'-hydroxybenzylidene)-cyclopentene-1,3-dione (KIH-200) and its 3'-methoxy and 3',5'-dimethoxy derivatives (KIH-201 and KIH-202) were investigated in vitro. These synthetic compounds were newly designed as non-nitro electron-affinic radiosensitizers and hypoxic cytotoxins on the basis of their high electron affinities and electrostatic potentials which were estimated by semiempirical molecular orbital calculation using the CNDO/2 program. The compounds were shown to have weak but apparent hypoxic cytotoxicity to FM3A cells from C3H mice. On irradiation in the presence of KIH-201 or KIH-202, hypoxic cells were more sensitive than aerobic cells at very low drug concentrations (below 10 microM), whereas KIH-200 was less effective.
Hitoshi Hori, G.-M. Pang, Kenzo Harimaya, Yoichi Iitaka and Seiichi Inayama : Isolation and Structure of Regelin and Regelinol, New Antitumor Ursene-type Triterpenoids From Tripterygium Regelii, Chemical & Pharmaceutical Bulletin, Vol.35, No.5, 2125-2128, 1987.
(要約)
Regelin (1a), a new antitumor ursene-type triterpenoid, the major principle, and another new ursene-type regelinol (1c) were isolated together with oleanene-type regelide (2) from the roots of Tripterygium regelii. The respective structures of 1a and 1c were determined, by ^1H-and ^<13>C-NMR spectrometric studies and X-ray analysis of 1a, to be the methyl ester of 3-oxo-22α-hydroxy-urs-12-en-30-oic acid and its 23-hydroxy analogue. 2 was found to be identical with wilforlide A or abruslactone A by comparing their physico-chemical data.
R.L. Stein, M. Strimpler, Hitoshi Hori and J.C. Powers : Catalysis by Human Leukocyte Elastase: Proton Inventory as a Mechanistic Probe, Biochemistry, Vol.26, No.5, 1305-1314, 1987.
(要約)
Proton inventories (rate measurements in mixtures of H2O and D2O) were determined for the human leukocyte elastase catalyzed hydrolyses of thiobenzyl esters and p-nitroanilides of the peptides MeOSuc-Val, MeOSuc-Alan-Pro-Val (n = 0-2), and MeOSuc-Alan-Pro-Ala (n = 1 or 2). The dependencies of k2/Ks on mole fraction of solvent deuterium for the p-nitroanilides are "dome-shaped" and were fit to a model that incorporates the mechanistic features of generalized solvent reorganization when substrate binds to enzyme and partial rate limitation of k2/Ks by physical and chemical steps [Stein, R. L. (1985) J. Am. Chem. Soc. 107, 7768-7769]. The proton inventories for the deacylation of MeOSuc-Val-HLE and MeOSuc-Pro-Val-HLE are linear while those for the deacylation of MeOSuc-Ala-Pro-Val-HLE and MeOSuc-Ala-Ala-Pro-Val-HLE are "bowl-shaped" and could be fit to a quadratic dependence of rate on mole fraction of deuterium. These results are interpreted to suggest that the correct operation of the catalytic triad is dependent on substrate structure. Minimal substrates, which cannot interact with elastase at remote subsites, are hydrolyzed via a mechanism involving simple general-base catalysis by the active site histidine and transfer of a single proton in the rate-limiting transition state. In contrast, tri- and tetrapeptide substrates, which are able to interact at remote subsites, are hydrolyzed by a more complex mechanism of protolytic catalysis involving full functioning of the catalytic triad and transfer of two protons in the rate-limiting transition state. Finally, the proton inventories for the deacylation of MeOSuc-Ala-Pro-Ala-HLE and MeOSuc-Ala-Ala-Pro-Ala-HLE are dome-shaped and suggest that the chemical events of acyl-enzyme hydrolysis are only partially rate limiting for these reactions and that some other physical step is also partially rate limiting.
R.L. Stein, M. Strimpler, Hitoshi Hori and J.C. Powers : Catalysis by Human Leukocyte Elastase: Mechanistic Insights into Specificity Requirements, Biochemistry, Vol.26, No.5, 1301-1305, 1987.
(要約)
Steady-state kinetic parameters were determined for the human leukocyte elastase catalyzed hydrolysis of a series of peptide-based thiobenzyl esters and p-nitroanilides. The peptide units are MeOSuc-Val, MeOSuc-Alan-Pro-Val (n = 0-2), and MeOSuc-Alan-Pro-Ala (n = 1 or 2). The results of this study suggest five important mechanistic features for HLE. Few important remote subsite contacts are established in the Michaelis complex. Full recognition and tight binding of the substrate occurs in the transition state for acylation. The P3-S3 interaction is critical during acylation. Subsite contacts are unimportant in deacylation. P1 specificity is regulated by peptide length. An important steady-state kinetic consequence of this specificity is that the rate-limiting step of kc for p-nitroanilide hydrolysis changes from acylation to deacylation as the peptide chain is lengthened.
K. Isobe, K. Yoshino, K. Ishibiki, O. Abe, Kenzo Harimaya, Hitoshi Hori, T. Shibata and Seiichi Inayama : Effect of SS-094 (Solcoseryl) on the Healing of Experimetnal Burns as Determined by Measuring Hydroxyproline Concentration in Tissue, Cytoprotection & Cytobiology (Excerpta Medica Current Clinical Practice Series 24, Amsterdam), Vol.1, 76-88, 1987.
160.
Hitoshi Hori, G.-M. Pang, Kenzo Harimaya, Yoichi Iitaka and Seiichi Inayama : The Absolute configuration of Regelidine, a Novel 6-Nicotinoyl Dihydroagarofuran Sesquiterpene Alkaloid From Tripterygium Regelii, Chemical & Pharmaceutical Bulletin, Vol.35, No.11, 4683-4686, 1987.
Y. Shibamoto, K. Ono, M. Takahashi, E. Kano, Hitoshi Hori, T. Shibata, Seiichi Inayama and M. Abe : An in vitro and in vivo Screeening System for New Hypoxic Cell Radiosensitizers Using EMT6 Cells, Japanese Journal of Cancer Research, Vol.77, 1027-1033, 1986.
163.
Kiyoshi Noguchi, Tatsuaki Hattori, Takashi Igarashi, Koichi Ueno, Tetsuo Satoh, Haruo Kitagawa, Hitoshi Hori, Tetsuichi Shibata and Seiichi Inayama : Effect on Hypoxic Cell Radiosensitizers on Glutathione Level and Related Enzyme Activities in Isolated Rat Hepatocytes, Life Sciences, Vol.37, No.7, 625-633, 1985.
(要約)
A comparative study of the effect of misonidazole and novel radiosensitizers on glutathione (GSH) levels and related enzyme activities in isolated rat hepatocytes was performed. Incubation of hepatocytes with 5 mM radiosensitizers led to a decrease in the intracellular GSH level. The most pronounced decrease in cellular GSH was evoked by 2,4-dinitroimidazole-1-ethanol (DNIE); after incubation for only 15 min, GSH was hardly detected. DNIE-mediated GSH loss was dependent upon its concentration. DNIE reacted with GSH nonenzymatically as well as with diethylmaleate, while misonidazole and 1-methyl-2-methyl-sulfinyl-5-methoxycarbonylimidazole (KIH-3) did not. Addition of partially purified glutathione S-transferase (GST) did not enhance DNIE-mediated GSH loss in a cell-free system. DNIE inhibited glutathione peroxidase (GSH-Px), GST, and glutathione reductase (GSSG-R) activities in hepatocytes, while misonidazole and KIH-3 did not. GSH-Px activity assayed with H2O2 as substrate was the most inhibited. Inhibition of GSH-Px activity assayed with cumene hydroperoxide as substrate and GST was less than that of GSH-Px assayed with H2O2 as substrate. GSSG-R activity was decreased by DNIE, but not significantly. Incubation of purified GSH-Px with DNIE resulted in a little change in the activity when assayed with H2O2 as substrate.
Yasuhiko Ohta, Carlos Jaime, Eiji Osawa, Yoichi Iitaka, Nobuko Shimizu, Shoko Nishihara, Tetsushi Ohsaka, Hitoshi Hori, Tetsuichi Shibata and Seiichi Inayama : A Note on the A-ring Conformation in 2-Chloro-1,2-dihydrosantonins, Chemical & Pharmaceutical Bulletin, Vol.33, No.1, 400-403, 1985.
(要約)
The half-chair and half-boat conformations observed by X-ray analysis for the A-ring of 2α- and 2β-chloro-1,2-dihydro-1-α-santonin 1,respecitvely, should be regarded as a deformed sofa conformation based on molecular mechanics.
Seiichi Inayama, Kenzo Harimaya, Nobuko Shimizu, Hitoshi Hori, Tamiko Ohkura and Takeshi Kawamata : Chemical Transformation of Aromatin into Pulchellin-A Formal Synthesis of Pulchellidine, Heterocycles, Vol.23, No.2, 377-381, 1985.
166.
Hitoshi Hori, A. Yasutake, Y. Minematsu and J.C. Powers : Inhibition of Human Leukocyte Elastase, Porcine pancreatic Elastase and Cathepsin G by Peptide Ketones. Peptides: Synthesis-structure-fuction, Proceedings of the Nineth American Peptide Symposium. C. M. Deber, V. J. Hruby and K. D. Kopple. Rockford, Pierce Chem. Co., 819-822, 1985.
167.
Susumu Hirose, Kazuyuki Takeuchi, Hitoshi Hori, Tadaaki Hirose, Seiichi Inayama and Yoshiaki Suzuki : Contact Points between Transcription Machinery and the Fibroin Gene Promoter Deduced by Functional Tests od Single-base Substitution Mutants, Proceedings of the National Academy of Sciences of the United States of America, Vol.81, No.5, 1394-1397, 1984.
168.
Seiichi Inayama, Kenzo Harimaya, Hitoshi Hori, Tamiko Ohkura, Takeshi Kawamata, M. Hikichi and T. Yokokura : Studies on Non-sesquiterpenoid Constituents on Gaillarida pulchella. II. Less Lipophilic Substances, Methyl Caffeate as an Antitumor Catecholic, Chemical & Pharmaceutical Bulletin, Vol.32, No.3, 1135-1141, 1984.
S. Suzuki, T. Inoue, T. Yasuda, T. Niwaguchi, Hitoshi Hori and Seiichi Inayama : Analysis of Methamphetamine in Human Hair by Mass Fragmentography, EISEI KAGAKU, Vol.30, No.1, 23-26, 1984.
170.
Takako Inoue, Yukiko Kanda, Tohro Kishi, Tokiyasu Sakai, Shinishi Suzuki, Tetsukichi Niwaguchi, Hitoshi Hori and Seiichi Inayama : Studies on Radioimmunoassay for Methamphetamine Extracted in Human Urine, Chemical & Pharmaceutical Bulletin, Vol.32, No.1, 1394-1397, 1984.
H. Baba, N. Aikawa, H. Amano, N. Ando, O. Abe, Hitoshi Hori, T. Shibata and Seiichi Inayama : New Extraction Method of Thromboxane B2 -etermination of TxB2 in Rats with Experimental Endotoxin Shock, Jpn. J. Inflammation, Vol.Suppl.4, 104-107, 1984.
172.
Seiichi Inayama, Nobuko Shimizu, Shoko Nishihara, Tetsushi Ohsaka, Hitoshi Hori, Tetsuichi Shibata, B. Andrzej Buda and Eiji Osawa : Equilibration of 2-Chloro-1,2-dihydrosantonin Conformers; A Theoretical Approach Using X-ray Diffraction and MO Calculations, Chemical & Pharmaceutical Bulletin, Vol.31, No.12, 4582-4585, 1983.
(要約)
2α-Chloro-1,2-dihydro-l-α-santonin (1) with the cyclohexenone A-ring of the ordinary half-chair form and the 2β-chloro isomer (2) with that of the unique half-boat from were found to equilibrate through acid-catalyzed epimerization and thermodynamic conformational change. In this, the former was favored over the latter by a free energy difference of 0.7-1.0 kcal/mol. The crystal structure and the half-chair conformation of the molecule of 1 were confirmed by X-ray diffraction methods. Semi-empirical (MNDO and CNDO) and ab initio molecular orbital calculations of these chloroenone conformers using input parameters obtained from their X-ray analytical data reproduced the observed conformational energy.
(キーワード)
2α- / 2β-chloro-1, 2-dihydro-l-α-santonin / half-chair / halfboat form / conformer / acid-catalyzed equilibration / stabilization energy / heat of formation / total energy / X線回折 (X-ray diffraction) / MO calculation (MNDO / CNDO / ab initio)
C. Murayama, Hitoshi Hori, T. Mori and Seiichi Inayama : Cytotoxic Properties of Hydroxylamino- and Amino-misonidazole, Possible Metabolic Products of Misonidazole, in Hypoxic HELA S3 Cells, Gan, Vol.74, No.5, 693-698, 1983.
(要約)
Misonidazole, a derivative of 2-nitroimidazole, has selective cytotoxic activity on hypoxic cells in addition to its radiosensitizing activity. This cytotoxicity is considered to be due to metabolic reduction of the drug. A possible metabolite seems to be hydroxylaminomisonidazole, an intermediate product derived via reduction of the nitro group. Authentic samples of hydroxylamino- and aminomisonidazole (a final reduction product) were synthesized and their cytotoxicity towards HeLa S3 cells was compared with that of misonidazole. After a 3-hr exposure to 1mM hydroxylaminomisonidazole under aerobic and hypoxic conditions, the surviving cell fractions were 0.18 and 0.0056, respectively. This represents a cytotoxicity five and 125 times greater, respectively, than that of misonidazole. Under the same conditions, aminomisonidazole showed no apparent cytotoxicity.
Seiichi Inayama, Kenzo Harimaya, Hitoshi Hori, Tamiko Ohkura, Takeshi Kawamata, Iwao Miura and Yoichi Iitaka : Structure and Conformation of a New Helenanolide, Pulchelloid C, Heterocycles, Vol.20, No.8, 1501-1506, 1983.
MANABU NODE, KIYOHARU NISHIDE, KEIICHIRO OHTA, KAORU FUJI, EIICHI FUJITA, Hitoshi Hori and SEIICHI INAYAMA : Hard Acid and Soft Nucleophile System. VI. A Convenient Synthesis of Alkylthiopolycyclic Aromatics with A Metal Halide and Thiol System, Chemical & Pharmaceutical Bulletin, Vol.31, No.2, 545-551, 1983.
(要約)
On treatment with a metal halide and alkanethiol system, polycyclic aromatics having various substituents such as hydroxy, alkoxy, phenoxy, acetoxy, and halogen were easily converted into alkylthiopolycyclic aromatics in high yields. The chemo and regioselectivity are described for the reaction of disubstituted naphthalenes.
(キーワード)
hard acid / soft nucleophile / aromatic sulfide / aluminum chloride / naphthalene / phenanthrene / anthracene / MO calculation
K. Isobe, K. Yoshino, K. Ishibiki, O. Abe, Kenzo Harimaya, T. Shibata, 堀 均, Seiichi Inayama : 組織ヒドロキシプロリン量よりみた実験的熱傷創の治癒状態とSS-094の創傷治癒に及ぼす影響, Cyto-protection & Biology, 63-73, 1983年.
177.
C. Murayama, Hitoshi Hori, Seiichi Inayama and T. Mori : A Comparative Study of the Radiosensitizing Effects and Cytotoxic Properties of Misonidazole and 2,4-Dinitroimidazole-1-ethanol in Hela S3 Cells, Gan, Vol.73, No.4, 588-591, 1982.
(要約)
Hypoxic HeLa S3 cells were used to compare the radiosensitizing and cytotoxic properties of misonidazole and 2,4-dinitroimidazole-1-ethanol. To achieve an enhancement ratio of 1.6 (C1.6), the required concentrations of 2,4-dinitroimidazole-1-ethanol and misonidazole were 0.15 and 2.4mM, respectively; at this ratio, the cytotoxicity values (1/To) of the two compounds were 0.054 hr-1 and 0.29 hr-1. These findings indicate that 2,4-dinitroimidazole-1-ethanol had a greater sensitizing effect at lower cytotoxicity, and that it is therefore the clinically more desirable radiosensitizing compound.
(キーワード)
Antineoplastic Agents / Cells, Cultured / Chemical Phenomena / 化学 (chemistry) / Dose-Response Relationship, Drug / Electrons / HeLa Cells / Humans / Misonidazole / Nitroimidazoles / Radiation-Sensitizing Agents
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 7152197
K. Isobe, K. Yoshino, S. Ishibiki, R. Abe, T. Shibata, Kenzo Harimaya, Hitoshi Hori and Seiichi Inayama : Effect of Solcoseryl® (SS-094) on Wound Healing of Dermal Burns in Ginea Pig (Based on Evaluation of Tissue Collagen), Pharmacometrics, Vol.24, No.4, 591-597, 1982.
181.
Seiichi Inayama, Nobuko Shimizu, Hitoshi Hori, Tetsushi Ohsaka, Tadaaki Hirose, Ttetsuichi Shibata and Yoichi Iitaka : Conformation of Chlorodihydrosantonins, Chemical & Pharmaceutical Bulletin, Vol.30, No.10, 3856-3859, 1982.
182.
C Shibata, Hitoshi Hori, Seiichi Inayama and T Mori : Radiosensitizing Effect of 2,4-Dinitroimidazole-1-ethanol and its Cytotoxicity in Hela S3 Cells, Strahlentherapie, Vol.157, No.7, 481-485, 1981.
(要約)
Using cultured HeLa S3 cells, the radiosensitizing and cytotoxic effects of newly synthesized derivatives of dinitroimidazole were investigated and compared with those of misonidazole. 2,4-Dinitroimidazole-1-ethanol radiosensitized hypoxic cells selectively. At 5 mM misonidazole, the enhancement ratio was 1.95; with 0.5 mM 2,4-dinitroimidazole-1-ethanol, almost the same enhancement could be obtained. This indicates that the radiosensitizing effect of the latter agent was about 10 times greater than that of misonidazole. However, its cytotoxicity was twice that of misonidazole under hypoxic conditions and there was no apparent differential cytotoxicity to hypoxic and aerobic cells.
Seiichi Inayama, Hitoshi Hori, Tetsushi Ohsaka, T. Mori and C. Shibata : An Approach to the Design and Synthesis of Mercaptoimidazole Derivatives Containing Oxidized Sulfur based on Electron Affinity Sensitization, Gan, Vol.72, No.1, 156-159, 1981.
(要約)
New mercaptoimidazole derivatives containing oxidized sulfur were designed and synthesized for screening with hypoxic radioresistant cells in place of nitroimidazoles, which are well-known radiosensitizing agents. The electron affinities of various mercaptoimidazole derivatives such as the sulfoxide (4) and sulfone (5) were estimated on the basis of the lowest unoccupied molecular orbital (LUMO) by using the CDNO/2 method. Compounds (4) and (5) thus designed were synthesized from the sulfide (3) by metaperiodate oxidation in 72% and 89% yields, respectively. No appreciable activity was observed with these compounds upon in vitro screening, probably due to their low water-solubility. Nevertheless, this approach may be useful for the development of improved radiosensitizers provided that mercaptoimidazole derivatives with adequate water-solubility can be obtained.
T. Mori, C. Shibata, Y. Ohizumi, H. Maezawa, S. Ushiro, Hitoshi Hori and S. Inayama : Hypoxic Cell Radiosensitizer in vitro, Japanese Journal of Cancer Clinics, Vol.27, No.12, 1447-1452, 1981.
185.
Hitoshi Hori, W. Kitajima, T. Shibata, Y. Ozawa and S. Inayama : An Approach to the Profiling Analysis of Urinary Prostaglandins by Simple Extraction Methods, Jpn. J. Inflammation, Vol.2, No.3, 247-250, 1981.
186.
Seiichi Inayama, Tetsushi Ohsaka, Tetsuichi Shibata, Tadaaki Hirose, Takeshi Kawamata, Hitoshi Hori and Yoichi Iitaka : Structure and Synthesis of Bissantanolides, Heterocycles, Vol.15, No.2, 861-864, 1980.
Seiichi Inayama, Hitoshi Hori, Tetsuichi Shibata, Yukio Ozawa, Keiichi Yamagami, Motoko Imazu and Hisako Hayashida : Simple and Rapid Separation of Certain Prostaglandins by Reversed-phase High-Performance Liquid Chromatography, Journal of Chromatography. A, Vol.194, No.1, 85-88, 1980.
Seiichi Inayama, Nobuko Shimizu, Tetuichi Shibata, Hitoshi Hori and Yoichi Iitaka : Unusual Bromination of Tetrahydro-(-)-α-santonins and New Santonin Isomers: X-ray Crystal and Molecular Structure of 2β, 14-Dibromo-4α,5β, 6β,11βH-tetrahydrosantonin, Journal of the Chemical Society. Chemical Communications, Vol.11, 495-496, 1980.
S. Inayama, Hitoshi Hori, T. Shibata, K. Yamagami and Y. Ozawa : Profiling Analysis of Prostaglandins on Bartter's Syndrome Using a New HPLC Separation Method, Jpn. J. Inflammation, Vol.1, No.2, 251-254, 1980.
190.
S. Ito, M. Numata, H. Ito, S. Inayama, Hitoshi Hori, M. Itoh, K. Saito, A. Aoyama and M. Iwaki : Time Course Effect of Anti-inflammatory Drugs on PGF2α and PGE1 as measured by GC/MS, Proc. Jpn. Soc. Med. Mass Spectrom., Vol.4, 263-268, 1979.
191.
M. Fujita, M. Node and Hitoshi Hori : Terpenoids. Part 39. Total Synthesis of Gibberellin A15 and Gibberellin A37, Journal of the Chemical Society, Perkin Transactions 1, Vol.6, 2144-2149, 1977.
192.
M. Node, Hitoshi Hori and M. Fujita : Terpenoids. XXXVII. Hypoiodite Reactions with 6-Hydroxy-17-norkaurene- and 7-Norgibberellane-derivatives, Chemical & Pharmaceutical Bulletin, Vol.24, No.9, 2149-2156, 1976.
193.
Manabu Node, Hitoshi Hori and Eiichi Fujita : Terpenoids. Part XXXVIII. Ring B Contraction of Kaurenolides and Related Compounds into Gibberellane-type Compounds, Journal of the Chemical Society, Perkin Transactions 1, Vol.20, 2144-2149, 1976.
Manabu Node, Hitoshi Hori and Eiichi Fujita : Demethylation of Aliphatic Methyl Ethers with a Thiol and Boron Trifluoride, Journal of the Chemical Society, Perkin Transactions 1, Vol.20, 2137-2140, 1976.
Manabu Node, Hitoshi Hori and Eiichi Fujita : Syntheses of Methyl Esters of Gibberellin A15 and Gibberellin A37, Journal of the Chemical Society. Chemical Communications, Vol.21, 898-899, 1975.
Phytyl quinols, namely acyclic tocopherols, are key intermediates of tocopherol biosynthesis,but their biological activities remain unclear. We therefore investigated the structure-activityrelationship of phytyl quinols to apply a chemical biosynthesis design for an antiatherosclerosisdrug based on isoprenomics. We have achieved the biosynthesis-oriented synthesis of α- andβ-phytyl quinol as an unnatural intermediate, other γ- and δ-phytyl quinol as a natural one. All fourphytyl quinols showed almost the same moderate inhibitory activity against low-density lipoproteinoxidation instead of their different degree of C-methylation with character different fromtocopherols. In vivo toxicities of phytyl quinols against chick embryo chorioallantoic membranevasculature were hardly observed. We proposed phytyl quinols were possible antioxidants in plantsand animals, like vitamin E.
Naoyuki Shimomura, Masataka Nagahama, Kenji Teranishi, Yoshihiro Uto and Hitoshi Hori : Introduction of Embryonic Chick Assay on Experiment Applying Nanosecond Pulse Electric Fields on Solid Tumor, IEEJ Transactions on Fundamentals and Materials, Vol.133, No.4, 231-232, 2013.
(要約)
Embryonic chick assay was introduced as a biological method for an experiment to apply a nanosecond pulse electric field on a tumor. Embryonic chick assay allowed many in vivo tests. Pulsed electric fields were applied on a solid tumor of the mouse breast adenocarcinoma cell, EMT6/KU, which was formed on the chorioallantoic membrane. We found that average weight of solid tumors applied pulsed electric fields was significantly smaller than that of controls.
(キーワード)
embryonic chick assay / in vivo / bioelectrics / nanosecond pulse electric fields / solid tumor
Eiji Nakata, Yoshijiro Nazumi, Yoshihiro Yukimachi, Yoshihiro Uto, Hiroshi Maezawa, Toshihiro Hashimoto, Yasuko Okamoto and Hitoshi Hori : Synthesis and photophysical properties of new SNARF derivatives as dual emission pH sensors, Bioorganic & Medicinal Chemistry Letters, Vol.21, No.6, 1663-1666, 2011.
(要約)
We report the synthesis and properties of two new seminaphthorhodafluor (SNARF) derivatives, SNARF-F and SNARF-Cl. Both these derivatives exhibit typical red shifts of absorption and fluorescence, and higher cell permeability as compared to traditional SNARF, while the pH-dependent dual-emission characteristics are well retained. In particular, the lower pK(a) (7.38) of SNARF-F makes it more suitable than traditional SNARF derivatives for intracellular applications.
Manda Sushma, Nakanishi Ikuo, Ohkubo Kei, Yoshihiro Uto, Kawashima Tomonori, Hitoshi Hori, Fukuhara Kiyoshi, Okuda Haruhiro, Ozawa Toshihiko, Ikota Nobuo, Fukuzumi Shunichi and Anzai Kazunori : Enhanced radical-scavenging activity of naturally-oriented artepillin C derivatives, Chemical Communications, No.5, 626-628, 2008.
(要約)
More than two-fold augmentation in the radical-scavenging activity of artepillin C could be achieved via altering the O H bond dissociation enthalpy of artepillin C by means of structural modifications.
Mok-Ryeon Ahn, Kazuhiro Kunimasa, Toshiro Ohta, Shigenori Kumazawa, Miya Kamihira, Kazuhiko Kaji, Yoshihiro Uto, Hitoshi Hori, Hideko Nagasawa and Tsutomu Nakayama : Suppression of tumor-induced angiogenesis by Brazilian propolis: Major component artepillin C inhibits in vitro tube formation and endothelial cell proliferation, Cancer Letters, Vol.252, No.2, 235-243, 2007.
(要約)
Propolis, a resinous substance collected by honeybees from various plant sources, possesses various physiological activities such as antitumor effects. We have previously shown that propolis of Brazilian origin was composed mainly of artepillin C and that its constituents were quite different from those of propolis of European origin. In this report, we examined an antiangiogenic effects of Brazilian propolis and investigated whether artepillin C was responsible for such effects. In an in vivo angiogenesis assay using ICR mice, we found that the ethanol extract of Brazilian propolis (EEBP) significantly reduced the number of newly formed vessels. EEBP also showed antiangiogenic effects in an in vitro tube formation assay. When compared with other constituents of EEBP, only artepillin C was found to significantly inhibit the tube formation of HUVECs in a concentration-dependent manner (3.13-50microg/ml). In addition, artepillin C significantly suppressed the proliferation of HUVECs in a concentration-dependent manner (3.13-50microg/ml). Furthermore, artepillin C significantly reduced the number of newly formed vessels in an in vivo angiogenesis assay. Judging from its antiangiogenic activity in vitro and in vivo, we concluded that artepillin C at least in part is responsible for the antiangiogenic activity of EEBP in vivo. Artepillin C may prove useful in the development of agents and foods with therapeutic or preventive activity against tumor angiogenesis.
Hitoshi Hori, Yoshihiro Uto and Eiji Nakata : Boron tracedrugs challenge for neutron dynamic therapy, Anticancer Research, Vol.32, No.6, 2235-2239, Jun. 2012.
(要約)
In this short review we describe our innovative boron tracedrugs and drugs for neutron dynamic therapy (NDT), as a newly emerging challenge beyond conventional drug treatments.
Hitoshi Hori, Yoshihiro Uto and Eiji Nakata : Medicinal electronomics bricolage design of hypoxia-targeting antineoplastic drugs and invention of boron tracedrugs as innovative future-architectural drugs, Anticancer Research, Vol.30, No.9, 3233-3242, Sep. 2010.
(要約)
We describe herein for the first time our medicinal electronomics bricolage design of hypoxia-targeting antineoplastic drugs and boron tracedrugs as newly emerging drug classes. A new area of antineoplastic drugs and treatments has recently focused on neoplastic cells of the tumor environment/microenvironment involving accessory cells. This tumor hypoxic environment is now considered as a major factor that influences not only the response to antineoplastic therapies but also the potential for malignant progression and metastasis. We review our medicinal electronomics bricolage design of hypoxia-targeting drugs, antiangiogenic hypoxic cell radiosensitizers, sugar-hybrid hypoxic cell radiosensitizers, and hypoxia-targeting 10B delivery agents, in which we design drug candidates based on their electronic structures obtained by molecular orbital calculations, not based solely on pharmacophore development. These drugs include an antiangiogenic hypoxic cell radiosensitizer TX-2036, a sugar-hybrid hypoxic cell radiosensitizer TX-2244, new hypoxia-targeting indoleamine 2,3-dioxygenase (IDO) inhibitors, and a hypoxia-targeting BNCT agent, BSH (sodium borocaptate-10B)-hypoxic cytotoxin tirapazamine (TPZ) hybrid drug TX-2100. We then discuss the concept of boron tracedrugs as a new drug class having broad potential in many areas.
Hideko Nagasawa, Yoshihiro Uto, Kenneth L. Kirk and Hitoshi Hori : Design of Hypoxia-Targeting Drugs as New Cancer Chemotherapeutics, Biological & Pharmaceutical Bulletin, Vol.29, No.12, 2335-2342, Dec. 2006.
(要約)
The tumor microenvironment is now recognized as a major factor that influences not only the response to conventional anti-cancer therapies but also helps define the potential for malignant progression and metastasis. In particular, hypoxia is now considered a fundamentally important characteristic of the tumor microenvironment. Furthermore, discovery of the hypoxia inducible factor 1alpha (HIF-1alpha) has led to a rapidly increasing understanding of the molecular mechanisms involved in tumor hypoxia. This in turn has led to the current extensive interest in the signal molecules related to tumor hypoxia as potential molecular targets for cancer therapeutics. In this paper we give an overview of recent advances in hypoxia research, including cancer treatments that target tumor hypoxia. Progress in the development of hypoxia-targeting drugs will be discussed, including antiangiogenic hypoxic cell radiosensitizers and hypoxic cytotoxins, hypoxia targeting boron carriers and p53-inhibiting bifunctional radiosensitizers. We will also review our own recent research results in these areas. For example, we have found that certain of the 2-nitroimidazole radiosensitizers and heterocycle-N-oxide hypoxic cytotoxins we developed have antiangiogenic activity and antimetastatic activity. We propose that these activities are based on the inhibition of signal transduction mediated by HIF-1alpha. The anti-tumor activities of hypoxia response are considered to be cytostatic (tumor dormancy-inducing) effects in contrast to cytotoxic DNA damaging effects. The combination of these cytostatic effects that are related to radiosensitization with the cytotoxic effects of radiation should improve the prognosis and QOL of patients receiving radiation and lead to an overall response to treatment. Based on these considerations, we developed the antiangiogenic hypoxic cell radiosensitizers, TX-1877, TX-1898 and the hypoxic cytotoxin TX-402 that inhibits the HIF-1alpha pathway We will also discuss our research involved with the development of other drugs to exploit tumor hypoxia, including a hypoxia-targeting boron carrier for boron neutron capture therapy (BNCT) and a p53 inhibiting radiosensitizer.
Ryu Tada, Hitoshi Hori, Shinichiro Masunaga, Yuko Kinashi, Koji Ono and Yoshihiro Uto : NDT-based Approach of Boron tracedrug UTX-51 to Glycated BSA as a AGE model., 15th International Congress of Radiation Research (ICRR2015), May 2015.
2.
Masataka Oita, Yoshihiro Uto, Hitoshi Hori, Masahide Tominaga and Motoharu Sasaki : Effects of Uncertainties of Radiation Sensitivity of Biological Modelling for Treatment Planning, AAPM 56th Annual Meeting, Jun. 2014.
3.
Yoshihiro Uto, Chiaki Abe, Toru Yoshitomi, Yukio Nagasaki, Yoshio Endo and Hitoshi Hori : Evaluation of in vivo antioxidative activity of O-TEMPO-RNP using our newly developed chicken egg assay, The 16th biennial meeting for the Society for Free Radical Research International (SFRRI), London, Sep. 2012.
4.
Masataka Nagahama, Naoyuki Shimomura, Yoshihiro Magori, Kenji Teranishi, Yoshihiro Uto and Hitoshi Hori : Development of Techniques Applying Nanosecond Pulse Electric Fields on Solid Tumor, Proceedings of the 2012 IEEE International Power Modulator and High Voltage Conference, 516-519, San Diego, Jun. 2012.
(要約)
Bioelectrics has become a notable field of pulsed power application at the present day. Cancer therapy using apoptosis with nanosecond pulsed electric fields (nsPEFs) has been studied in laboratories. We introduced embryonic chick assay to study in vivo. The embryonic chick assay has several advantages and is categorized as in vivo experiment. Many samples can be prepared with low cost by the assay. This study adopted EMT6/KU (mouse breast adenocarcinoma cells) as tumor cells, which formed a solid tumor on the chorioallantoic membrane (CAM) of a fertilized hen egg in the assay. Nanosecond pulsed electric fields with width of 1.5 ns were applied on the solid tumor on CAM with needle electrodes. Stainless wires of 1 mm in diameter were used as the needle electrodes. The two or four needle electrodes were used in this experiment. The solid tumor was dissected and extracted from the egg on a three days after application of nsPEF and was then weighed. The controls of tumor sample were also prepared without nsPEF application. Significance test on the weight of tumors was adopted to determine the effect of nsPEFs application. The difference of weight between the tumor with nsPEF application and control was significantly confirmed. The difference enlarged with the charging voltage of pulsed power generator.
Yoshihiro Magori, Seiji Ohta, Tatsuya Kageyama, Kenji Teranishi, Naoyuki Shimomura, Yoshihiro Uto and Hitoshi Hori : In Vivo Experiment of Applying Nanosecond Pulsed Electric Fields on Solid Tumor, Proceedings of the 18th IEEE International Pulsed Power Conference, 1237-1241, Chicago, Jun. 2011.
Yoshihiro Uto, Chiaki Abe, Toru Yoshitomi, Yukio Nagasaki, Yoshio Endo, Eiji Nakata and Hitoshi Hori : In vivo antioxidative activity of radical-containing-nanoparticle (RNP) in fertilized chicken egg assay, International Conference on Biomaterials Science 2011, Tsukuba, Mar. 2011.
7.
Yoshihiro Uto, Chiaki Abe, Takashi Nakae, Yoshio Endo, Masahide Tominaga, Hiroshi Maezawa, Eiji Nakata and Hitoshi Hori : Systems biology-based drug design of sugar-hybrid hypoxic cell radiosensitizers using the tumor-implantable chick embryo model, PACIFICHEM 2010, Honolulu, Dec. 2010.
8.
Hitoshi Hori, Eiji Nakata, Masato Koizumi, Yohei Yamashita and Yoshihiro Uto : Boron tracedrug: Design, synthesis and pharmacological activity of phenolic BODIPY-containing antioxidants as traceable next-generation drug model, International Society on Oxygen Transport to Tissue 2010 (ISOTT 2010), Ascona, Switzerland, Jul. 2010.
9.
Yoshihiro Uto, Ryo Tanaka, Kenta Ohnaka, Yuki Ohta, Kazufumi Yazaki, Naoyuki Umemoto, Eiji Nakata and Hitoshi Hori : Prenylated acylphloroglucinol derivatives: Isoprenomics-based design, syntheses and antioxidative activities, International Society on Oxygen Transport to Tissue 2010 (ISOTT 2010), Ascona, Switzerland, Jul. 2010.
10.
Eiji Nakata, Yukimachi Yoshihiro, Nazumi Yoshijiro, Abe Chiaki, Yoshihiro Uto, Hiroshi Maezawa and Hitoshi Hori : Design of a Bioreductively-Activated Fluorescent pH Probe for Tumor Hypoxia Imaging, SJBC2009, Tokyo, Sep. 2009.
11.
Hitomi Nakashima, Kazuhiro Ikkyu, Kouichiro Nakashima, Keiichiro Sano, Yoshihiro Uto, Eiji Nakata, Hideko Nagasawa, Hiroshi Sugimoto, Yoshitsugu Shiro, Yoshinori Nakagawa and Hitoshi Hori : Design of novel hypoxia-targeting IDO hybrid inhibitors conjugated with an unsubstituted L-Trp as an IDO affinity moiety, ISOTT2008 (2008 International Society on Oxygen Transport to Tissue Conference), Sapporo, Aug. 2008.
(要約)
Indoleamine 2,3-dioxygenase (IDO) catalyzes the initial and rate limiting step of L-Tryptophan (L-Trp) catabolism in the kynurenine pathway. IDO expresses in many neoplasms, and play a role in immunosuppression. IDO is activated only reductive or hypoxic conditions, because the heme iron of active site is easily oxidized to its inactive form. Hypoxic neoplastic cells indicate chemo- and radioresistances and also cause neoplasm' progression, invasion and metastasis. Previously we focused to the mode of activation of IDO only in hypoxic and reductive condition to design hypoxia-targeting IDO hybrid inhibitors, which constituted an IDO inhibitor 1-methyl-tryptophan (1MT) and hypoxic cytotoxin tirapazamine (TPZ), such as TX-2236, TX-2235, TX-2228 and TX-2234. They were good hypoxia-targeting IDO inhibitors. 1MT-TPZ hybrids inhibited IDO uncompetitively. Thus, it suggests that 1MT-TPZ hybrids were able to bind the only enzyme-substrate complex, not to bind the active site. In this study, we present here the novel design and syntheses of hypoxia-targeting IDO hybrid inhibitors conjugated with an unsubstituted L-Trp as an IDO affinity moiety without inhibitor 1MT such as L-Trp-TPZ hybrids 1 (n = 4, monoxide), 2 (n = 4, dioxide), 3 (n = 5, monoxide), and 4 (n = 5, dioxide). These L-Trp-TPZ hybrids were synthesized from 3-chlorinated TPZ monoxide with various length alkyldiamines to give the corresponding amine derivatives, which were conjugated with Boc-L-Trp to give the L-Trp-TPZ monoxide hybrids 1 and 3. L-Trp-TPZ dioxide hybrids 2 and 4 were also synthesized from the corresponding TPZ-dioxide trifluoroacetamide intermediates. L-Trp-TPZ hybrids were good competitive inhibitors of IDO. Thus, it suggests that these L-Trp-TPZ hybrids were able to bind the active site. In conclusion, we discovered IDO hybrid inhibitors conjugated with an unsubstituted L-Trp as an IDO affinity moiety. Additionally we are under evaluation of their hypoxic cytotoxicity to develop hypoxia-targeting IDO hybrid inhibitors as our final goal.
12.
Yoshihiro Uto, Chiaki Abe, Yoshio Endo, Eiji Nakata, Hideko Nagasawa and Hitoshi Hori : Development of an in vivo evaluation system of antioxidants for their vascular protective activities using the chick embryo chorioallantoic membrane, ISOTT2008 (2008 International Society on Oxygen Transport to Tissue Conference), Sapporo, Aug. 2008.
(要約)
The blood vessel is one of the most important organs in the oxygen transport, and the drug discovery research of clinical-use antioxidants, which may control various vascular disorders caused by the oxidative stress, is extremely important. In fact, the in vitro evaluation system of antioxidants being mainly used now, however, has a problem of no appreciable in vivo redox status of the antioxidants. We, therefore, tried the development of an in vivo model for the evaluation of antioxidants for their vascular protective activities using the chick embryo having many advantages, such as handy, quick, cheap, and possible high-throughput screening. The topical administration (doses from 150 to 350 g) of 2,2'-azobis(2-methylpropionamidine) dihydrochloride (AAPH), a water-soluble prooxidant, to the chorioallantoic membrane (CAM) of 6-days chick embryos, resulted in their vascular injuries of percentage from 25 to 100% dose dependently, which were correlated with their increased fatal rate of the percentage from 29 to 86%. Artepillin C, a potent natural antioxidant, showed a weak vascular injury (26% at 10 g). These results suggest that immature developing CAM vasculature might have low tolerance to oxidative injury. In the case of 12-days chick embryos, AAPH (doses from 0.3 to 7 mg) induced their vascular injuries (20 to 80%) against the CAM veins and venous capillaries without their fatal damage. Artepillin C, with its dose of up to 30 g, did not show the chick embryo's venous injury, and pre-treatment with artepillin C dose-dependently protected the CAM venous and venous capillary injuries induced by the post-administration of 3 mg of AAPH: 1 g artepillin C decreased 60% of venous injury. These results suggest that artepillin C efficiently protected the AAPH-induced vascular injury of chick embryonic CAM. In conclusion, artepillin C might be a promising clinical-use antioxidant for prevention of vascular disease.
13.
Hitoshi Hori, Yoshihiro Uto, Daisuke Koyama, Mamoru Otsuki, Naoki Otomo, Tadashi Shirai, Chiaki Abe, Eiji Nakata and Hideko Nagasawa : A chemical biosynthesis design for antiatherosclerosis drug by acyclic tocopherol intermediate analogue based on ``isoprenomics'', ISOTT2007 (2007 International Society on Oxygen Transport to Tissue Conference), Uppsala, Sweden, Aug. 2007.
(要約)
Tocopherols constitute members of the vitamin E group and act as a potent lipid peroxidation inhibitor against for cell membrane such as blood vessels. Phytyl quinols, namely acyclic tocopherols, are key intermediate of tocopherol biosynthesis, but their biological activities remain unclear. We therefore investigated the structure-activity relationship of phytyl quinols to apply chemical biosynthesis design for an antiatherosclerosis drug. We have achieved the biosynthesis-oriented design and synthesis of alpha- (TX-2254) and beta- (TX-2247) phytyl quinol as an unnatural intermediate, other gamma- (TX-2242) and delta- (TX-2231) phytyl quinol as a natural one. Free radical reactivity was determined by using DPPH radical. Inhibitory activity of human low density lipoprotein (LDL) oxidation was measured by TBARS assay. Free radical reactivity of TX-2242 (EC50 = 24.6 microM) and TX-2231 (24.2 microM) were equal with DL-alpha-tocopherol (22.3 microM), whereas TX-2254 (38.0 microM) and TX-2247 (30.2 microM) showed lower reactivity than DL-alpha-tocopherol. TX-2242 (IC50 = 61 microM) and TX-2231 (69 microM) also showed almost same LDL antioxidant activity as DL-alpha-tocopherol (64 microM), while TX-2254 (149 microM) and TX-2247 (254 microM) showed lower antioxidant activity than DL-alpha-tocopherol. We suggested from these results that the cyclization of phytyl quinol to tocopherol is not indispensable for the appearance of the potent antioxidant activity. We also have found an interesting negative correlation between methylation and antioxidant activity of phytyl quinols. We will present ex vivo antioxidant activity for blood vessels in the chick embryo chorioallantoic membranes (CAM) as an alternative in vivo model.
14.
Junichi Morishige, Kenji Fukuzawa, Hideko Nagasawa, Hitoshi Hori and Akira Tokumura : Lysophosphatidic acid produced by lysophospholipase D in hen egg white induces blood vessel formation on hen chorioallantoic membrane, 3rd International Conference on phospholipase A2 and lipid mediators, Sorrento, May 2007.
15.
Hitoshi Hori, Yoshihiro Uto, Hideko Nagasawa, Shutaro Ae, Daisuke Koyama, Naoki Otomo, Mamoru Otsuki and Takashi Tuji : Design and synthesis of LDL oxidation inhibitor based on isoprenomics, XIII Biennial Meeting of the Society for Free Radical Research International, Davos, Aug. 2006.
16.
Hitoshi Hori, Hideko Nagasawa, Yoshihiro Uto, Masamitsu Hiraoka, Keiko Goto, Hitomi Nakashima, Satoru GOTO and Shin-ichiro Masunaga : Design, synthesis, and pharmacokinetics of hypoxic tumor-targeting boron carriers, IMEBORON12, Sendai, Sep. 2005.
(要約)
Purpose: The presence of hypoxic tumor cells is also associated with resistance to boron neutron capture therapy (BNCT). To accomplish the effective boron carrier for BNCT, we have recently developed 2-nitroimidazole-sodium borocaptate-10B (BSH) conjugates, TX-2016, 2017, 2018, 2041, 2042, and 2060, that is, hybrids that have both hypoxic tumor cell g-ray-sensitizing unit, 2-nitroimidazoles, and thermal neutron-sensitizing unit, BSH [1]. Furthermore, In this communication, to improve the hypoxia-targating character, we report the design, synthesis, and pharmacokinetics of hypoxic cytotoxin-hybrid boron carriers, TX-2091, 2095, 2097, 2100, 2124 and 2125, having hypoxic cytotoxin moiety, such as 2-quinoxalinecarbonitrile-1,4-di-N-oxide (TX-402) and tirapazamine (TPZ), and BSH via some carbon chain linker as a pharmacokinetic controlling unit. Result: TX-2016, 2017, and 2018: tertamethylammonium salts of alkyl sulfides of BSH. TX-2041, 2042 and 2060: sodium salts of an alkyl sulfide or a dialkyl sulfonium derivatives of BSH. They were synthesized from their corresponding haloacetylcarbamoyl-2-nitroimidazoles[2] and BSH. To synthesize hypoxic cytotoxin conjugate, 3-clorinated hypoxic cytotoxin (TX-402 or TPZ) was condensed with ethyl-, propyl-, or hexyl-alcoholamine to obtain the corresponding hydroxyalkylamino derivatives. They were treated with bromoacetyl isocyanate followed by disodium (2-cyanoethyl)-thio-undecahydro-closo-dodecaborate(2-) to yield cyanoethyl sulfonium compounds, TX-2095, 2100, 2110, 2124, and 2125. Among the 2-nitroimidazole conjugates, TX-2041 has the most favorable pharmacokinetic characteristics with higher tumor affinity of 10B during neutron beam irradiation. In addition, TX-2041 showed a significantly higher radiosensitizating effect with reactor thermal neutron beams than that of BSH. Cyanoethyl sulufonium derivative, TX-2060 showed better tumor affinity of 10B and more cytotoxicity after irradiation with reactor thermal neutron beams than that of TX-2041. Furthermore, TX-2100 achieved significantly higher tumor affinity than 2-nitoroimidazoles, TX-2041 and 2060, and also showed cytotoxicity comparable to TX-2060. In conclusion, these hypoxic tumor trageting cytotoxin-hybrid 10B-carriers , such as TX-2100, with the potential tumor affinty of10B, are expected to have clinical applications as promising boron carriers for use in BNCT.
17.
Shinichi Nakayama, Hideko Nagasawa, Tomoya Fujita, Toshihiro Hashimoto, Yoshinori Asakawa, Yoshihiro Uto, Hitoshi Hori and Douglas A. Kuntz : Synthesis of new GMII mannosidase inhibitors, diversity-oriented 5-substituted swainsonine analogues, via stereoselective Mannich reaction, 230th American Chemical Society Meeting & Exposition, Washington, D.C., Aug. 2005.
(要約)
A potent Golgi a-mannosidase (GMII) inhibitor, (-)-swainsonine, exhibits pleiotropic effects such as anticancer activities including metastasis and immunomodulatory effects. To develop more potent immunomodulatory anticancer agents and gain insight on the molecular bases of these activities, we synthesized 5-substituted swainsonine analogues stereoselectivity using our developed diversity-oriented modification reaction (ref.1). We have developed amine acetal (1) as a versatile intermediate for the preparation of 5a-(3) and 5b-acyl (4). Swainsonine analogues were synthesized by stereoselective Mannich reaction of an in situ generated (-)-swainsonine iminium ion intermediate (2) followed by their epimerization reaction. Their carbonyl groups were reduced by Pd-catalyzed hydrogenation to afford 5a-(5a) and 5b-alkyl analogues(5b). Some of these new 5a-substituted swainsonine analogues possessing aromatic group showed more potent a-mannosidase inhibitory activities than swainsonine itself. To gain insight on the molecular bases of these activities, we also analyzed the crystal structures of Dorosophila GMII in complex with some of these swainsonine analogues.
18.
Hitoshi Hori, Hideko Nagasawa, Yoshihiro Uto, Cheng-Zhe Jin, Ayako Tanaka, Mariko Shimamura, Yoshio Takeuchi, Kenneth L. Kirk and Seiichi Inayama : Design of 2-methylene-4-cyclopentene-1,3-dione-containing azomycin derivatives as antiangiogenic hypoxic cell radiosensitizers, 96th American Association for Cancer Research Annual Meeting, Anaheim, Apr. 2005.
19.
Hitoshi Hori, Yoshihiro Uto, Mitsutoshi Sakakibara and Hideko Nagasawa : Artepillin C isoprenomics; first total synthesis of artepillin C and medicinal approach for artepillin C analogues, 3rd International Symposium on Natural Drugs, Napoli, Italy, Oct. 2003.
Yoshihiro Uto, Hideko Nagasawa and Hitoshi Hori : Phenolic natural product isoprenomics; first total synthesis of artepillin C and medicinal approach for artepillin C analogues, 226th ACS National Meeting, New York, Sep. 2003.
Hitoshi Hori, Hideko Nagasawa and Yoshihiro Uto : Structure-based design of the antitumor 2-hydroxyarylidene-4-cyclopentene-1,3-dione TX-1123 as a protein tyrosine kinase inhibitor having low mitochondrial toxicity, 3rd International Conference on Inhibitors of Protein Kinases and Workshop on Phosphoryl-Transfer Mechanisms, Warsaw, Poland, Jun. 2003.
Hideko Nagasawa, Naoko Mikamo, Yoshimi Nakajima, Hideki Matsumoto, Yoshihiro Uto and Hitoshi Hori : TX-402: Hypoxic cytotoxin as an inhibitor pf hypoxia-inducible factor 1 pathway, The Tumor Microenvironment and Its Inpact on Cancer Therapies 8th International Workshop, Miami, Florida, May 2003.
Hitoshi Hori, Hideko Nagasawa, Yoshihiro Uto, Kazuto Ohkura, Y. Takeuchi, K.L. Kirk, H. Miyataka and T. Sato : Design of selective arachidonic acid cascade inhibitors of low-mitochondrial-toxic PTK inhibitor TX-1123 analogues based on their COX-1, 2 and 5-LO inhibitory activities, The 8th International Workshop on the Tumor Microenvironment and Its Impact on Cancer Therapies, Miami, Florida, USA, May 2003.
Atsushi Umemoto, M.A. Momen, J. Honda, J.-W. Zhu, Megumi Yukawa, Yoshihiro Uto, Hideko Nagasawa, Hitoshi Hori and Yasumasa Monden : Possible potent activity of 7-Oac-UDCA, a novel compound, in the prevention of colon cancer, 18th World Congress of Digestive Surgery (International Society for Digestive Surgery), 9th Hong Kong International Cancer Congress, Hong Kong, China, Dec. 2002.
Hitoshi Hori, Soko Kasai, Hideko Nagasawa, Yoshihiro Uto, T. Inomata, S. Oka and S. Inayama : TX-1877: Designed antimetastatic hypoxic cell radiosensitizer, The 6th China-Japan Joint Symposium on Drug Design and Development (Asian Federation for Medicinal Chemistry), Dalian, China, Oct. 2001.
S.B. Mohamad, Hitoshi Hori, Hideko Nagasawa, Kenji Usui and Yoshihiro Uto : Characterization of human Gc protein-derived macrophage activation factor (GcMAF) and its functional role in macrophage tumoricidal activity, 29th ISOTT (International Society on Oxygen Transport to Tissue) annual meeting, Philadelphia, PA, USA, Aug. 2001.
T. Satoh, E. Miyataka, E. Jayachandran, K.L. Kirk, Y. Takeuchi and Hitoshi Hori : Cancer Cell-Killing Agent with an Extremely Wide anticarcinogenic Spectrum and Almost No Side Effects, AACR(American Associationfor Cancer Research) 92nd Annual Meeting, New Orleans, LA, USA, Mar. 2001.
29.
Y. Nakagawa, T. Suda and Hitoshi Hori : The difference of bleaching of platonin derivatives by active oxygen species, 10th Biennial Meeting of the International Society for Free Radical Research (SFRR 2000), Kyoto, Oct. 2000.
30.
Hitoshi Hori, Yoshihiro Uto, Akihiko Hirata, Mitsutoshi Sakakibara, Masaki Ishibashi and Hideko Nagasawa : Structure-based elucidation nof antioxidative and antiperoxidative effects with 2,4,6-trisubstituted phenols, 10th Biennial Meeting of the International Society for Free Radical Research (SFRR 2000), Kyoto, Oct. 2000.
(要約)
2,4,6三置換フェノール類の抗酸化作用美ついての構造活性相関を論述した．
31.
Hideko Nagasawa, Fumi Nagura, Saharuddin B. Mohamad, Ji-Wen Zhu, Yoshihiro Uto, T. Hashimoto, Yoshimori Asakawa and Hitoshi Hori : Apoptosis Induc-tion of Cyto-chalasins Iso-lated from The Japanese Fungus Daldinia Ver-nicaso for Human Colon Cancer Cell, Third Interna-tional Congress on Phytomedi-cine, Munich, Germany, Oct. 2000.
Hitoshi Hori, Soko Kasai, Hideko Nagasawa, Mao Yamashita, Yoshihiro Uto, I Ota and N Matsuura : Design, Synthesis, and Activities of Antimetastatic Hypoxic Cell Radiosensitizer TX-1877 and its Analogues Inhibiting Endothelial Cells Migration, 11th Int'l Conference of Chemical Modifiers of Cancer Treatment, Banff, Alberta, Canada, Oct. 2000.
Hitoshi Hori, Soko Kasai, Hideko Nagasawa, Mao Yamashita, Yoshihiro Uto, I Ota and N. Matsuura : Design, synthesis, and activities of antimetastatic hypoxic cell radiosensitizer TX-1123 and its analogues inhibiting endotherlial cells migration, Tumor Physiology and Cancer Treatment, 8th International Conference-Chemical Modifiers of Cancer Treatment Series, Banff, Alberta, Canada, Oct. 2000.
Yoshihiro Uto, Akihiko Hirata, Masaki Ishibashi, Hideko Nagasawa and Hitoshi Hori : Design and synthesis of 2,6-diprenyl-4-iodophenol TX-1952 having a novel and potent anti-peroxidative activity, ISOTT(International Society on Oxygen Transport to Tissue) 2000 annual meeting, Nijmegen, Netherlands, Aug. 2000.
Hideko Nagasawa, Mao Yamashita, Naoko Mikamo, Mariko Shimamura, S. Oka, Yoshihiro Uto and Hitoshi Hori : Design, synthesis and biological activities od antiangiogenic hypoxic cytotoxin, triazine-N-oxide derivatives, ISOTT2000 (International Society on Oxygen Transport to Tissue), Nijmegen, Netherlands, Aug. 2000.
S.B. Mohamad, Hideko Nagasawa, Yoshihiro Uto and Hitoshi Hori : Tumor cell alpha-N-acetylgalactosaminidase activity and its involvement in GcMAF-related macrophage activation, ISOTT2000 (International Society on Oxygen Transport to Tissue), Nijmegen, Netherlands, Aug. 2000.
Hitoshi Hori, J.-W. Zhu, Hideko Nagasawa, Fumi Nagura, S.B. Mohamad, Yoshihiro Uto and Kazuto Ohkura : Brefeldin A: Toward new Differentiation and Apoptosis Inducers, Fourth Australia/Japan Joint Symposium on Drug Design and Development, Werribee, Victoria, Australia, May 2000.
(要約)
ブレフェルジンAの分化誘導作用と抗腫瘍活性及びアポトーシス誘導作用について発表した．
38.
Soko Kasai, Hideko Nagasawa, Mao Yamashita, Yoshihiro Uto, I Ota, N Matuura and Hitoshi Hori : Design, Synthesis and effect of hypoxic cell radiosensitizer TX-1877 and its derivatives as endothelial cells migration inhibitor, MILLENNIAL WORLD CONGRESS OF PHARMACEUTICAL SCIENCES, San Francisco, Apr. 2000.
I Ota, Soko Kasai, Hitoshi Hori and N Matuura : A novel function of TX-18777, a hypoxic cell radiosensitizer, as anti-angiogenic agent, Keystone Symposium on Experimental and Clinical Regulation of Angiogenesis, Salt Lake City, Mar. 2000.
40.
Hitoshi Hori, Yoshihiro Uto, Masaki Ishibashi, Akihiro Hirata and Hideko Nagasawa : Artepillin C: Molecular modeling and synthetic approach, 2nd International Conference on Food Factors, Kyoto, Dec. 1999.
(要約)
プロポリス成分，アルテピリンCの全合成を完成させてた．
41.
J.-W. Zhu, Hideko Nagasawa, Fumi Nagura, S.B. Mohamad, Yoshihiro Uto, Kazuto Ohkura and Hitoshi Hori : 4-epi-Brefeldin A: probing new structural requirements of brefeldin A for inducing activity of differentiation and apoptosis in human colon cancer cell HCT 116, AACR-NCI-EORTC International Conference: Molecular Tragets and Cancer Therapeutics, Discovery, Development, and Clinical Validation, Washington, DC, USA, Nov. 1999.
Satoru GOTO, Hitoshi Hori, Zenei Taira and Hiroshi Terada : Leukotriene antagonists: Biological activities and their dynamic structures, The 2nd China/Japan Symposium on Drug Design and Development, Beijing, Jan. 1991.
Yoshihiro Uto, ryota Takeuchi, Yoshinori Nakagawa, Keiji Hirota, Hiroshi Terada, Shinya Onizuka, Kentaro Kubo, Daisuke Kuchiike, Mette Martin, Toshio Inui, Yoshio Endo and Hitoshi Hori : Development of Immunomodulatory Cancer Therapy Based on Gc protein-derived Macrophage Activating Factor (GcMAF), 7th International Symposium on Nanomedicine, Nov. 2013.
(キーワード)
癌免疫療法 (cancer immunotherapy) / GcMAF
15.
Yoshihiro Uto, Yoshio Endo, Hiroshi Sato and Hitoshi Hori : Development of antimetastatic hypoxic cytotoxin TX-2137 targeting for Akt/protein kinase B, 72th Annual Meeting of the Japanese Cancer Association, Oct. 2013.
Yoshihiro Uto and Hitoshi Hori : beta-Galactosidase treatment is common modification method of three major subtypes of Gc protein to GcMAF in vivo, 71th Annual Meeting of the Japanese Cancer Association, Sep. 2012.
Yoshihiro Uto, Onizuka Shinya and Hitoshi Hori : Effect of the Macrophage Activating Factor Precursor (preGcMAF) on Phagocytic Activation of Mouse Peritoneal Macrophages, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
36.
Chiaki Abe, Yoshihiro Uto, Yoshio Endo, Hiroshi Maezawa, Shin-ichiro Masunaga and Hitoshi Hori : Evaluation of the In vivo Radiosensitizing Activity of Etanidazole Using Tumor-bearing Chick Embryo, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
Hitoshi Hori, Eiji Nakata and Yoshihiro Uto : Design of boron tracedrug phenolic BODIPY-containing antioxidants as autopsy/virtopsy imaging agents, 第69回日本癌学会学術総会, Sep. 2010.
54.
Yoshihiro Uto, Chiaki Abe, Eiji Nakata and Hitoshi Hori : SAR analysis of electron-rich polyyne analogues of FTY720 based on in vivo chick embryo antiangiogenic assay, 第69回日本癌学会学術総会, Sep. 2010.
We designed hypoxia-targeting drugs, azomycin-2-methylene-4-cyclopentene-1,3-dione derivatives(McAZs) as anti-angiogenic hypoxic cell radiosensitizers which are incorporated a chiral 1-hydrophobic alkyloxy-or aryloxy-ethyl group substituent on an electron-deficient azomycin moiety, and a 2-methylene-4-cydopentene-1,3-dione. To evaluate McAZs as anti-angiogenic hypoxic cell radiosensitizer, we examjned radiosensitizing effects, hypoxic cytotoxicities anti-angiogenic activities, and PTK inhibitory activities. TX-2036,was expectedly a promising and potent EGFR kinase inhibitor, having an IC50 value of lower than 1.84 μM and also was a Flt-1 kinase inhibitor with an IC50 value of lower than 18.4 μM. TX-2036 also showed a high radiosensitizing effect with an ER of 1.79 at l µM in EMT6/KU cell. There was also good hypoxic cytotoxicity, and very strong anti-angiogenic activities showing 64% inhibition at 1 μg/CAM by CAM assay and an IC50 of 0.81 μM to RLE cells.
Yoshihiro Uto, Hitoshi Hori, 乾 利夫 and 久保 健太郎 : Pharmaceutical composition and method of preparing same, US 13/988,376 (Sep. 2012), US8,747,919 (Jun. 2014).