Potential of α-amino alcohol p-boronophenylalaninol as a boron carrier in boron neutron capture therapy, regarding its enantiomers

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.

Journal of Cancer Research and Clinical Oncology


Usefulness of combined treatment with mild temperature hyperthermia and/or tirapazamine in the treatment of solid tumors: its independence of p53 states

本論文では，p53正常または変異がん細胞を移植した担癌マウスモデルにおける放射線または化学療法による治療において，マイルドハイパーサーミア単独あるいはTPZとの併用処理が，p53ステータスにかかわらず特にQ細胞に対して有効であることを示した．


Preparation of Gc Protein-derived macrophage Activation Factor (GcMAF) and its Structural Characterization and Biological Activities.

本論文において，Gcタンパク質からマクロファージ活性化因子(GcMAF)へのプロセシング反応を解明するため，糖鎖修飾と生物活性の相関を検討した．その結果GcMAFのマクロファージ活性化作用においてN-acetylgalactosamine糖鎖が重要であることを明らかにした．


TX-1123: an antitumor 2-hydroxyarylidene-4-cyclopentene-1,3-dione as a protein tyrosine kinase inhibitor having low mitochondrial toxicity

A series of 2-hydroxyarylidene-4-cyclopentene-1,3-diones were designed, synthesized, and evaluated with respect to protein tyrosine kinase (PTK) inhibition, mitochondrial toxicity, and antitumor activity. Our results show that the cyclopentenedione-derived TX-1123 is a more potent antitumor tyrphostin and also shows lower mitochondrial toxicity than the malononitrile-derived AG17, a potent antitumor tyrphostin. The O-methylation product of TX-1123 (TX-1925) retained its tyrphostin-like properties, including mitochondrial toxicity and antitumor activities. However, the methylation product of AG17 (TX-1927) retained its tyrphostin-like antitumor activities, but lost its mitochondrial toxicity. Our comprehensive evaluation of these agents with respect to protein tyrosine kinase inhibition, mitochondrial inhibition, antitumor activity, and hepatotoxicity demonstrates that PTK inhibitors TX-1123 and TX-1925 are more promising candidates for antitumor agents than tyrphostin AG17.

本論文において，チロシンキナーゼ阻害を介した抗腫瘍剤をめざして2-hydroxyarylidene-4-cyclopentene-1,3-dione誘導体の分子設計·合成及び生物活性の検討を行った．その結果，AG17と比較してミトコンドリア毒性の低い，強力な抗腫瘍性チロフォスチンであるTX-1123の開発に成功したことを示した．


An Unusual Sesquiterpene Derivative from Ferula Kuhistanica

Ferula Kuhistanicaより単離したセスキテルペン誘導体の構造決定と活性を明らかにした．


First Total Synthesis of Artepillin C Established by o,o'-Diprenylation of p-Halophenols in Water

本論文では，プロポリスの生理活性成分として注目される，artepillin Cの初の全合成を達成した．


Tumor cell alpha-N-acetylgalactosaminidase activity and its involvement in GcMAF-related macrophage activation

本論文では，GcMAFのマクロファージ活性化機構を明らかにするため，腫瘍細胞株におけるalpha-N-acetylgalactosaminidase(alpha-NaGalase)がエキソ型であり，正常細胞に比べて活性化されていることを示した．また，alpha-NaGalaseによってGcMAFのマクロファージ活性化能が阻害されることを明らかにした．

Comparative Biochemistry and Physiology. Part A: Molecular & Integrative Physiology


Design and synthesis of 2,6-diprenyl-4-iodophenol TX-1952 with a novel and potent anti-peroxidative activity

本論文において，抗酸化剤をめざして2,4,6-三置換フェノール誘導体の分子設計·合成を行い，これらの生物活性を調べたところ，TX-1952は非常に強い抗酸化活性を有することを明らかにした．


Design, synthesis and biological activities of antiangiogenic hypoxic cytotoxin, triazine-N-oxide derivatives

本論文において，hypoxic cytotoxinであるTPZをリードとして，トリアジンまたはキノキサリンーNーオキシド類の分子設計を行い，生物活性を調べた．その結果，Hypoxic cyto-toxinはp53非依存的アポトーシス誘導作用を有し，さらに強い血管新生阻害作用を示すことを明らかにした．


Design and Synthesis of Antiangiogenic/Heparin-Binding Arginine Dendrimer Mimicking the Surface of Endostatin

本論文では，血管新生阻害タンパク質のエンドスタチン構造ミミックとしてアルギニンデンドリマーの分子設計，合成，生物活性の検討を行った．その結果，我々の開発したTX-1944は，強い血管新生阻害作用を示し，ヘパリン結合能を有することを明らかにした．

Bioorganic & Medicinal Chemistry Letters


Comparision of Hypoxic Cell Radiosensitizers, KIN-804, KIN-844, KIN-806 and TX-1877, on Brain and Liver Metabolizing Capacities in Mice Bearing Ehrlich Ascites Carcinoma

本論文において，エーリッヒ癌細胞移殖マウスを用いて，低酸素細胞放射線増感剤KIN-804，844，806及びTX-1877の脳及び肝臓の代謝能におよぼす影響を検討した．まず，神経毒性の指標としてアセチルコリンエステラーゼ(AChE)阻害作用について検討し，上記の増感剤はいずれもMISOと異なり毒性を示さないことを見出した．また，KIN-806とTX-1877は，抗酸化システムであるGSH及びSOD活性を上昇させた．以上より，KIN-806及びTX-1877は放射線照射によって減少する抗酸化システムを回復させるユニークで毒性の低い低酸素細胞放射線増感剤として有望な化合物であることを明らかにした．


Synthesis and Biological Activities of Fluorinated Chalcone Derivatives

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.


GcMAFはヒト単球のFcレセプター介在性ロゼット形成能を顕著に増強させるが，スーパーオキサイド産生能にはほとんど影響を及ぼさない


Modification of Cell Response to Insulin by Membrane-acting Agents in Rat White Adipoctes: Analysis of Structural Features by Compytative Simulation

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.


Inhibition of green tea catechins against the growth of cancerous human colon and heptatic epithelial cells

The ability of (-)-epicatechin (EC), (-)-epigallocatechin (EGC) and (-)-epigallocatechin gallate (EGCG) to inhibit the growth of HCT 116 colorectal and Hep G2 hepatocellular carcinoma cells was examined by MTT and clonogenic assays (CA). The respective catechins inhibited the growth of HCT 116 more strongly than Hep G2. In MTT assay, IC(50) values of EGC and EGCG against HCT 116 grew smaller on prolongation of the exposure times of the cells to the catechins. In CA, however, these two catechins had IC(50) values ranging between 7.6+/-0.4 and 11.2+/-0.5 microM against the same cells regardless of the exposure times. EC showed much weaker growth inhibitions relative to the two aforementioned catechins.

本論文において，種々の緑茶カテキン類の抗腫瘍効果を示した．


EVALUATION OF THE POTENTIAL OF PARA-BORONOPHENYLALANINOL AS A BORON CARRIER IN BORON NEUTRON CAPTURE THERAPY, REFERRING TO THE EFFECT ON INTRATUMOR QUIESCENT CELLS

C57BL mice bearing EL4 tumors and C3H / He mice bearing SCC VII tumors received 5-bromo-2'-deoxyuridine (BrdU) continuously for 5 days via implanted mini-osmotic pumps to label all proliferating (P) cells. Three hours after oral administration of l-p-boronophenylalanine-(10)B (BPA), or 30 min after intraperitoneal injection of sodium borocaptate-(10)B (BSH) or l-p-boronophenylalaninol (BPA-ol), a newly developed (10)B-containing alpha-amino alcohol, the tumors were irradiated with thermal neutron beams. For the combination with mild temperature hyperthermia (MTH) and / or tirapazamine (TPZ), the tumors were heated at 40 degrees C for 30 min immediately before neutron exposure, and 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 MN and apoptosis frequency in total (P + Q) tumor cells were determined from tumors that were not pretreated with BrdU. Without TPZ or MTH, BPA-ol increased both frequencies most markedly, especially for total cells. However, as with BPA, the sensitivity difference between total and Q cells was much larger than with BSH. On combined treatment with both MTH and TPZ, this sensitivity difference was markedly reduced, similarly to when BPA was used. MTH increased the (10)B uptake of all (10)B-compounds into both tumor cells. BPA-ol has good potential as a (10)B-carrier in neutron capture therapy, especially when combined with both MTH and TPZ.

本論文では，腫瘍移殖担癌マウスモデルにおいて，p-boronophenylalaninol(BPA-ol) の熱中性子照射における効果を明らかにした．BPA-olは腫瘍への取り込みが高い優れたボロンキャリアであり，マイルドハイパーサーミアまたはTPZとの併用でさらに増感されることを示した．


Radiosensitization effect by combination with paclitaxel in vivo, including the effect on intratumor quiscent cells

To evaluate the radiosensitization effect on solid tumors upon combination treatment with paclitaxel (TXL), including the effect on intratumor quiescent (Q) cells. Mice bearing SCC VII or EL4 solid tumors received 5-bromo-2'-deoxyuridine (BrdU) continuously for 5 days to label all proliferating (P) cells. The mice then received gamma-irradiation with or without tirapazamine (TPZ) at various time points after TXL administration. Another group of mice received a series of test doses of gamma-rays while alive or after tumor clamping to obtain hypoxic fractions (HFs) in the tumors at various time points after TXL administration. Immediately after irradiation, the tumor cells were isolated and incubated with a cytokinesis blocker. The micronucleus (MN) frequency in cells without BrdU labeling (Q cells) was determined using immunofluorescence staining for BrdU. Meanwhile, 6 h after irradiation, the tumor cells were isolated from the solid tumors in another group of mice, and the apoptosis frequency in Q cells was also determined with immunofluorescence staining for BrdU. The MN and apoptosis frequency in total (P + Q) tumor cells were determined from the tumors that were not pretreated with BrdU. For the measurement of the HFs, the MN or apoptosis frequency of Q cells was then used to calculate the surviving fraction of Q cells from the regression line for the relationship between the MN or apoptosis frequency and the surviving fraction of total tumor cells. In both SCC VII and EL4 tumors, maximum values of mitotic index (MI) and apoptosis frequency were observed 9 and 24 h after TXL administration, respectively. However, on the whole, the apoptosis frequency for SCC VII was very low. gamma-Irradiation 9 h after TXL administration induced significant radiosensitization effects on the total cells of both tumors. Irradiation at 60 h had a more significant effect on total cells of EL4 tumor, but no significant effect on total cells of SCC VII tumor. Combined treatment with TXL induced no radiosensitization effect on Q cells in either tumor. The effect on Q cells was observed only after TPZ was administered. The HF of total cells in EL4 tumors decreased significantly 60 h after TXL administration. No radiosensitization effect upon combination treatment with TXL is induced in Q tumor cells. However, the effect on P cells is produced by irradiation at the time when the maximum values of MI are induced following TXL administration. In addition, for tumors that are susceptible to apoptosis after TXL administration alone, irradiation at the time of sufficient reoxygenation in tumors after TXL administration produces a greater radioenhancement effect on P cells.

本論文では，腫瘍移殖担癌マウスモデルにおいて，TPZ存在または非存在下での放射線処理にpaclitaxel(TXL)を併用した場合の影響について解析した．その結果，TXLはP細胞において増感作用を示すことを明らかにした．

International Journal of Radiation Oncology*Biology*Physics


New Antimetastatic Hypoxic Cell Radiosensitizers: Design, Synthesis, and Biological Activities of 2-Nitroimidazole-acetamide, TX-1877, and its Analogues

We designed, based on the molecular orbital (MO) calculation, synthesized, and evaluated the biological activities of the new antimetastatic hypoxic cell radiosensitizer, 2-nitroimidazole-acetamide, TX-1877, and its analogues. Each analogue has an electron-affinic imidazole group, an acetamide group and a certain hydrophilic group to control its biological effect, toxicity, and pharmacokinetics. In in vitro radiosensitization assay, most TX-1877 analogues, which have an electron affinity (EA) of more than 0.9 eV and partition coefficient (P) of more than 0.021, showed satisfactory enhancement ratios (ER > 1.60) at doses of I mM. On the other hand, imidazole analogues, such as TX-1908 (EA = 0.67 eV), TX-1910 (EA = -0.34 eV) and TX-1931 (EA = -0.37 eV), which have low electron affinities, had an ER of 1.31 or less. TX-1877 and KIN-806 effectively inhibited tumor regrowth when administered with irradiation in vivo at a dose of 0.4 mg/g. Tumor lung metastasis was inhibited by treatment with either TX-1877 or KIN-806 without irradiation at a dose of 0.4 mg/g. TX-1877 reduced markedly the mean number of metastatic lung nodules in comparison with KIN-806. Moreover, TX-1877 and KIN-806 enhanced macrophage and helper T lymphocyte infiltration for 3 weeks after drug treatment. TX-1877 shows a high EA value and has the C2 of HOMO localizing on N-methylamide and the C2 of LUMO localizing on 2-nitroimidazole group. The MO data might be useful for designing a bifunctional hypoxic cell radiosensitizer. TX-1877 and its analogues are potential antimetastatic hypoxic cell radiosensitizers, which would improve the efficiency of radiotherapy and quality of life in cancer treatment.

本論文において，多機能性低酸素細胞放射線増感剤TX-1877及びそのアナログの分子設計を行い，これらの低酸素放射線増感作用，血管新生阻害作用，転移抑制作用について，構造活性相関を行い，それらの有効性を示した．


Exploiting tumor hypoxia in the treatment of solid tumors

Japanese Journal of Hyperthermic Oncology


Change in oxygenation status in intratumour total and quienscent cells following γ-ray irradiation,tirapazamine administration, cisplatin injection and bleomycin treatment

C3H/He mice bearing SCC VII tumours received 5-bromo-2'-deoxyuridine (BrdU) continuously for 5 days via implanted mini-osmotic pumps to label all proliferating (P) cells. The mice then received gamma-ray irradiation, or administration of tirapazamine (TPZ), cisplatin or bleomycin. At various time points after each treatment, tumour-bearing mice were irradiated with a series of test doses of gamma-rays, while alive or after being killed, to obtain hypoxic fractions (HFs) in the tumours. Immediately after gamma-ray test irradiation, the tumours were excised, minced and trypsinized. Tumour cell suspensions obtained were incubated with cytochalasin-B, a cytokinesis blocker, and the micronucleus (MN) frequency in cells without BrdU labelling (i.e. quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. MN frequency in the total (P + Q) tumour cells was determined from the tumours that were not pre-treated with BrdU. MN frequency of BrdU-unlabelled cells was then used to calculate the surviving fraction of the unlabelled cells from the regression line for the relationship between the MN frequency and the surviving fraction of total tumour cells. TPZ and cisplatin reduced the HF after treatment, especially in Q cells, and this tendency was particularly marked with TPZ. In contrast, bleomycin increased the HF after treatment. Both reoxygenation following gamma-ray irradiation or bleomycin treatment and a subsequent return to pre-treatment levels of HF following TPZ or cisplatin treatment (rehypoxiation) occurred more rapidly in total (P + Q) cells than in Q cells. Based on our previous report that total (P + Q) and Q cells within this tumour have large acutely and chronically HFs, respectively, we conclude that acute hypoxic cells play a major role in reoxygenation and rehypoxiation in SCC VII tumours.

本論文では，腫瘍移殖担癌マウスモデルに対して，放射線，TPZ，シスプラチン投与，ブレオマイシン処理を行った場合の，総腫瘍細胞またはQ細胞における酸素化ステータスの変化を調べた．この結果TPZはQ細胞における低酸素分画を減少させることを示した．


Changes in the Sensitivity of Intratumor Cells during Fractionated Tirapazamine Administration

Mice bearing solid tumors received 10 intraperitoneal administrations of 5-bromo-2'-deoxyuridine (BrdU) to label the proliferating (P) tumor cells. Then, as a priming treatment, tirapazamine (TPZ) was intraperitoneally administered. Further, 0 through 48 h later, the tumor-bearing mice received TPZ again at various doses. The tumor cells were isolated and incubated with a cytokinesis blocker. The micronucleus (MN) frequencies in cells with and without BrdU labeling, which were regarded as P and quiescent (Q) cells at the priming treatment, respectively, were determined using immunofluorescence staining for BrdU. The MN frequency in the total (P + Q) tumor cells was determined from the tumors that were not pretreated with BrdU. In addition, P cell ratios in the tumors at the second treatment were determined using immunofluorescence staining for P cell nuclear antigen. In each cell fraction, the longer the interval between the two treatments, the higher was the sensitivity to TPZ, except 1 h after the priming treatment. More than 24 h later, total and P cells, especially P cells, showed significantly higher sensitivity to TPZ than in the case of a single TPZ treatment. The longer the period between the two TPZ treatments, the lower was the P cell ratio at the second treatment. These findings were thought to indicate that the use of TPZ in the treatment of solid tumors causes a shift from the P to the Q state in vivo.

本論文では，腫瘍移殖担癌マウスモデルにおいて，TPZの分割投与により，その感受性を増すことを示した．この際，増殖期細胞(P細胞)がQ細胞にシフトしていることも明らかにした．


Usefulness of Tirapazamine as a Combined Agent in Chemoradiation and Thermo-chemoradiation Therapy at Mild Temperatures: Reference to the Effect on Intratumor Quiescent Cells

C3H / He mice bearing SCC VII tumors received 5-bromo-2'-deoxyuridine (BrdU) continuously for 5 days via implanted mini-osmotic pumps to label all proliferating (P) cells. The mice then received one of six different DNA-damaging agents with or without mild temperature hyperthermia (40 degrees C, 30 min, MTH). These agents were adriamycin (ADM), mitomycin C (MMC), cyclophosphamide (CPA), bleomycin (BLM), cisplatin (CDDP), and tirapazamine (TPZ). After the drug treatment, the tumor-bearing mice were irradiated with a series of doses of gamma-rays. Immediately after irradiation, 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. The MN frequency in the total (P + Q) tumor cells was determined from the tumors that had not been pretreated with BrdU. MTH significantly increased the MN frequency of total cells in tumors irradiated with gamma-rays combined with CPA, BLM, CDDP or TPZ, and that of Q cells in tumors irradiated with gamma-rays combined with BLM or TPZ. The sensitivity difference in the MN frequency between total and Q tumor cells was significantly decreased by the combination with TPZ. TPZ combined with radiotherapy and TPZ combined with thermo-radiotherapy at mild temperatures appear to be promising modalities for sensitizing tumor cells in vivo, including Q tumor cells.

本論文では，腫瘍移殖担癌マウスモデルにおいて，TPZは種々の化学療法剤に比べて，放射線との併用，あるいはマイルドハイパーサーミア及び放射線との併用により，Q細胞に対して優位に増感作用を示すことを明らかにした．以上により，このような癌治療法において，TPZを併用することは有効であることを示した．


Studies of Methyl 2-Nitroimidazole-1-acetohydroxamate (KIN-804)1: Effect on Free Radical Scavenging System in Mice Bearing Ehrlich Ascites Carcinoma

Methyl 2-nitroimidazole-1-acetohydroxamate (KIN-804) is a 2-nitroimidazole derivative containing a hydroxamate side chain designed to enhance the radiosensitization response of hypoxic cells. The possible sensitization of tumor tissue by KIN-804 can be evaluated through investigation of the levels of the free radical scavengers; namely, glutathione (GSH) and its complex enzyme system including glutathione reductase (GR) and glutathione peroxidase (GSH-Px), as well as glucose-6-phosphate dehydrogenase (G-6-PD). Female albino mice were inoculated with Ehrlich carcinoma in the thigh. Administration of KIN-804 (i.p. 80 mg/kg body weight) was carried out 20 min before localized irradiation of 10 Gy. The data revealed that KIN-804 administration, followed or not by gamma irradiation, resulted in a significant decrease in GSH content in tumor tissues associated with inhibition in GR and G-6-PD activities. Blood GSH-Px was enhanced in tumor inoculated mice and the administration of KIN-804 returned it to the normal value. These changes were more noticeable in tumor bearing mice exposed to both KIN-804 and irradiation.

本論文では，エーリッヒ癌細胞移殖マウスにおいて，低酸素放射線増感剤KIN-804単独または放射線との併用により，生体のラジカル消去システムであるグルタチオン含量が，腫瘍内で抑制されることを示した．これは，グルタチオンレダクターゼ及びglucose-6-phosphate dehydrogenaseの阻害と関連していると考えられた．


Studies of Methyl 2-Nitroimidazole-1-acetohydroxamate (KIN-804)2: Effect on Certain Antioxidant Enzyme Systems in Mice Bearing Ehrlich Ascites Carcinoma

In order to decrease toxicity and/or increase radiosensitizing activity, a new 2-nitroimidazole derivative, methyl 2-nitroimidazole-1-acetohydroxamate (KIN-804), was synthesized to solve the problem of tumor hypoxia. Evaluation of the efficiency of KIN-804 was carried out through studying the antioxidant enzyme system: The superoxide dismutase (SOD), catalase and lipid peroxide levels provide a rough index of the balance between free radical generation and scavenging. Female albino mice were inoculated with Ehrlich ascites carcinoma (EAC) in the thigh. The administration of KIN-804 (i.p. 80 mg/kg body weight) was carried out 20 min before localized irradiation of 10 Gy. In general, the data revealed that KIN-804 administration, followed or not by gamma irradiation, exerted significant inhibition of SOD and catalase activities accompanied by a significant increase in lipid peroxide level in tumor-bearing mice.

本論文では，エーリッヒ癌細胞移殖マウスにおいて，低酸素放射線増感剤KIN-804単独または放射線との併用により，生体のラジカル消去システムであるSOD及びカタラーゼ が抑制されることを示した．これにより，生体の脂質過酸化が増大した．


Elucidation of Strict Structural Requirements of Brefeldin A as an Inducer of Differentiation and Apoptosis

Brefeldin A (BFA) can induce a wide variety of human cancer cells to differentiation and apoptosis and is in development as an anticancer agent. To elucidate structural requirements for cytotoxicity and induction of differentiation and apoptosis, BFA was structurally modified into various derivatives including 4-epi-BFA in this study. Their inducing activities of apoptosis were evaluated with their cytotoxicities, DNA fragmentation and morphological changes in human colon cancer cell HCT 116. The cytotoxicity of 4-epi-BFA (TX-1923) (IC50 = 60 microM) was 300 times lower than that of BFA (IC50 = 0.2 microM). Furthermore, 4-epi-BFA induced DNA fragmentation and apoptotic morphological changes at much higher concentrations (70 and 50 microM, respectively) than BFA (0.11 and 0.36 microM, respectively). These results indicated that the configuration of 4-hydroxyl group of brefeldin A plays a key role in the cytotoxicity and induction of apoptosis. On the other hand, 7-O-acetyl-BFA, 4-O-acetyl-BFA, and 4,7-di-O-acetyl-BFA exhibited potential activities in cytotoxicity and inducibility of apoptosis. We suggested that the structural determinants for BFA include the moiety of the Michael acceptor, the conformational rigidity of the 13-membered ring, and the configuration of 4-hydroxyl group.

本論文では，brefeldin A(BFA)をリードとして種々のアナログの分子設計，合成を行い，それらの分化誘導作用を介したアポトーシス誘導効果に関して構造活性相関の解析を行った．


Apoptosis induction in HCT116 cells by cytochalasins isolated from the fungus Daldinia vernicosa

本論文において，Daldinia Venicosaより単離，構造決定したサイトカラシン類の大腸がん由来HCT116細胞に対するアポトーシス誘導作用を明らかにし，これらの構造活性相関について論述した．


Analyses of Insulin-Potentiating Fragments of Human Growth Hormone by Computative Simulation; Essential Unit for Insulin-Involved Biological Responses

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.


The immunopotentiation effects of the bifunctional radiosensitizer KIN-806 in comparison with its analogs KIN-804 and KIN-844

本論文において，新規な多機能性低酸素細胞放射線増感剤 KIN-806，KIN-804及び844が腫瘍移殖担癌マウスモデルにおいて，強い放射線増感効果を有すること，さらに特にKIN-806投与群においては，マクロファージ/T-リンパ球の腫瘍への浸潤が活性化され，肺転移の抑制が認められたことから，KIN-806には免疫賦活，転移抑制効果があることも明らかにした．


Lung metastasis suppression of the bifunctional new radiosensitizer KIN-806

本論文では，新規な多機能性低酸素細胞放射線増感剤 KIN-806が腫瘍移殖担癌マウスモデルにおいて，強い放射線増感効果のほかに，肺転移を抑制する効果があることを示した．

International Journal of Molecular Medicine


Potentially Lethal Damage Repair by Total and Quiescent Tumor Cells Following Various DNA-damaging Treatments

本論文では，マウス移殖腫瘍モデルにおいて，全腫瘍細胞とQ細胞に対する放射線，TPZ投与，シスプラチン投与などのDNA障害性処理による潜在的致死損傷の回復(PLDR)に関する解析を行った．Q細胞では全細胞に比べて，いずれの処理においても強い回復能を示した． ただし，TPZ処理の場合には，他の処理に比べて両者の細胞ともに回復能の低下が見られた．


The Design and Synthesis of 2-nitroimidazoles Compounds as a Hypoxic Cell Radiosensitizer and Their Anti-angiogenic Activity


Modification of tirapazamine-induced cytotoxycity in combination with mild hyperthermia and/or nicotinamide: reference to effect on quiescent tumour cells

本論文では，マウス移殖腫瘍モデルにおいて，TPZのQ細胞および低酸素細胞に対する細胞毒性が．マイルドハイパーサーミアまたはmicotinamideを併用することにより，増感されることを明らかにした．


Effects of Sho-saiko-to, a Japanese Herbal Medicine, on Hepatic Fibrosis in Rats

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.


Analysis of Structure-Permeability Correlation of Nitrophenol Analogues in Newborn Rat Abdominal Skin Using Semiempirical Molocular Orbital Calculation

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.


Enhancement Effect of Methyl α-D-Glucoside for Inhibitory Effects of Antioxidants on ADP/Fe2+-Induced Lipid Peroxidation in Rat Liver Mitochondria

本論文では，ラット肝ミトコンドリアにおいて，ADP/Fe2+で誘導される膜脂質過酸化に対する，種々の抗酸化剤の効果をmethyl -D-glucosideが増強することを示した．本論文では，ラット肝ミトコンドリアにおいて，ADP/Fe2+で誘導される膜脂質過酸化に対する，種々の抗酸化剤の効果をmethyl -D-glucosideが増強することを示した．

Advances in Experimental Medicine and Biology


Retinyl palmitate reduces hepatic fibrosis in rats induced by dimethylnitrosamine or pig serum


Enhancement of Cisplatin Sensitivity of Quiescent Cells in Solid Tumors by Combined Treatment with Tirapazamine and Low-Temperature Hyperthermia

本論文では，マウス移殖腫瘍モデルにおいて，tirapazamine(TPZ)とマイルドハイパーサーミアの併用により，固形腫瘍に存在すると考えられる休止期の細胞(Q細胞)のシスプラチンに対する化学療法感受性が増感することを明らかにした．


TX-1877: DESIGN, SYNTHESIS, AND BIOLOGICAL ACTIVITIES AS A BRM-FUNCTIO HYPOXIC CELL RADIOSENSITIZER

2-Nitroimidazole acetamide TX-1877 and its derivatives (TX-1877 analogs) were designed, synthesized, and evaluated by their in vitro and in vivo radiosensitization, tumor growth control, suppression of lung metastasis, and immunopotentiation, as biological response modifier (BRM)-functional hypoxic cell radiosensitizers. TX-1877 analogs were designed and synthesized in our laboratory. In vitro radiosensitizing ability was estimated using EMT6/KU cells under hypoxic conditions. In vivo radiosensitization, antimetastasis, and immunopotentiation were evaluated using female C3H/He mice bearing the SCCVII tumor. Days (15 or 10) after the inoculation of 10(5) SCCVII tumor cells into the hinder thigh, a drug (0.4 mg/g) was administered i.p. and local irradiation of 30 Gy was given at 30 min after its administration. Tumor growth was observed for 20 days and mice were euthanized to count the number of metastatic nodules on the surface of the lungs. Tumor tissues were extirpated and stained by the ABC method at 1, 2, and 3 weeks after treatment for immunological evaluation. Novel types of bifunctional radiosensitizers, TX-1877 and its analogs possessing BRM-functions (i.e., antimetastatic and immunopotentiation effects) were developed. In vitro radiosensitizing abilities of TX-1877 and its analogs, with their partition coefficient values of more than 0.050, were comparable to misonidazole (MISO) at their doses of 1 mM. Tumor regrowth was suppressed evidently 20 days after the treatment in the irradiated group with TX-1877 (TX-1877 plus R) and with KIN-806 (KIN-806 plus R). The former group reduced markedly the mean number of metastatic lung nodules regardless of radiation therapy. TX-1877 and KIN-806 plus R induced helper T lymphocytes. The TX-1877, TX-1877 plus R, KIN-806, and KIN-806 plus R enhanced macrophage infiltration for 3 weeks after treatment. TX-1877 is an excellent BRM-functional hypoxic cell radiosensitizer, expected to be useful for clinical use.

本論文において，多機能性低酸素細胞放射線増感剤 TX-1877の分子設計·合成を行い，このものが強い放射線増感効果のほかに，転移抑制作用とマクロファージの浸潤を活性化する免疫賦活作用を有することを明らかにした．

International Journal of Radiation Oncology*Biology*Physics


Appplicability of Combination with Tirapazamine in Boron Neutron Capture Therapy

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.


Dynamic Structural Features of Macrocyclic Cytochalasin Analogues Responsible for Their Hexose Transport Inhibition

Journal of the American Chemical Society


Effect of Extracellular Calcium on the Intracellular Calcium Level of Newborn Rat Skin Basal Cells

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.


Effects of Bioreductive Agents, Tirapazamine and Mitomycin C, on Quiescent Cell Populations in Solid Tumors, Evaluated by Micronucleus Assay

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.


Respiratory activities of liver mitochondria, isolated from freshwater turtle chinemys revesii as an experimental anoxia-tolerant model system, determined by mitochondrial modifiers

本論文では，酸欠耐性モデルとして，クサガメ(Chinemys reevesii)より単離した肝ミトコンドリア(TLM)を用いて，ミトコンドリア修飾剤による呼吸活性の解析を行った．その結果，TLMの呼吸活性が低いために，基質酸化速度が遅いことが高い酸欠耐性に寄与していることを明らかにした．


Bifunctional hypoxic cell radiosensitizer KIH 896の肺転移抑制作用に関する実験的研究 --- -C57BL/6マウス·ルイス肺癌細胞を用いたKIN806との比較検討-


Synthesis and Activity of Brefeldin a Analogs as Inducers of Cancer Cell Differentiation and Apotosis

Bioorganic & Medicinal Chemistry Letters


Design, Synthesis, and Biological Activity of Anti-angiogenic Hypoxic Cell Radiosensitizer Haloacetylcarbamoyl-2-nitroimidazoles

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.


Stablilities of the fluorescent SH-reagent eosin-5-maleimide and its adducts with sulfhydryl compounds

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.

Biochimica et Biophysica Acta (BBA) - General Subjects


A Convenient Screening Test for Hypoxic Cell Radiosensitizers/Cytotoxins

A convenient in vitro screening test using E. coli B/r for evaluating a variety of hypoxic cell radiosensitizers/hypoxic cell cytotoxins has been developed for the initial selection of candidates in medicinal/organic chemistry laboratories. E. coli cells were used for convenience since: (1) the bacterium is grown using commercially available broths, where it multiplies rapidly, and requires little specialized equipment for growth and handling. (2) More is known about the genetics and biochemistry of the radiation damage to these cells and their repair than any other organism.

本論文において，大腸菌を利用した低酸素細胞放射線増感剤の簡便な活性試験法を開発し，これを種々の低酸素細胞放射線増感剤に適用して活性を比較し，その有用性を示した．


In Vivo Radiosensitizing Effect of Nitroimidazole Derivative KIN-804

In vivo characteristics of 2-nitroimidazole-1-methylacetohydroxamate (KIN-804), which is a newly developed hypoxic cell radiosensitizer, are presented. The toxicity, pharmacokinetics, and radiosensitizing effect of KIN-804 were studied by in vivo experiments using C3H/He mice bearing the SCC-VII tumor. Results were compared with misonidazole (MISO). LD50(7) of KIN-804 and MISO were 3200 mg/kg and 2000 mg/kg, respectively. The peak of concentrations of KIN-804 in the tumor occurred 20 min after intraperitoneal injection and reached about 62% of the maximum concentration in the blood. The concentrations in brain and sciatic nerve were very low and clearance from sciatic nerve was rapid. Enhancement ratios of KIN-804 calculated using the growth delay method were 1.22, 1.50 and 1.71 at doses of 50, 100, and 200 mg/kg, respectively, compared with 1.36 for MISO at a dose of 100 mg/kg. In the TCD50 assay, enhancement ratios at a dose of 200 mg/kg were 1.69 for KIN-804 and 1.52 for MISO, respectively. KIN-804 is a promising radiosensitizer since it shows less toxicity and higher radiosensitizing activity than MISO.

本論文では，低酸素細胞放射線増感剤KIN-804(論文11)に関して，マウスによるin vivo活性試験と毒性試験を行った結果，既存の増感剤，ミソニダゾール(MISO)よりも高い活性を有し，毒性は低いことを明らかにした．また脳への移行も少なく，新規低酸素細胞放射線増感剤として非常に有望であることも示した．

International Journal of Radiation Oncology*Biology*Physics


Quantitative structure-activity relationships of benzamide derivatives for anti-leukotriene activities

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.


Radiosensitizing Effect of a 2-Nitroimidazole Hydroxamate (KIN-804) to Murine Tumors

ヒドロキサム酸型ニトロイミダゾールKIN-804の実験動物腫瘍に対する放射線増感効果

We studied the toxicity, pharmacokinesis and radiosensitizing effect of a newly developed 2-nitroimidazole-1-methylacetohydroxamate (KIN-804) in C3H/He mice bearing SCC-VII tumor. They were compared with misonidazole. LD50/7 of KIN-804 and misonidazole were 3200 and 2000 mg/kg. The concentration of KIN-804 in the brain and sciatic nerve was at a very low level and its clearance from the sciatic nerve was rapid. Enhancement ratios calculated using the growth delay method were 1.50 for KIN-804 and 1.36 for misonidazole respectively when they were administered by intraperitoneal injection with a dose of 100 mg/kg. KIN-804 was considered to be a promising radiosensitizer because it obtained less toxicity and a higher radiosensitizing activity than misonidazole.

本論文では，先に開発した低酸素細胞放射線増感剤KIN-804(論文11) に関して，マウスによるin vitro及びvivo活性試験と毒性試験を実施し，MISOよりも高い活性を有し，毒性は低いことを明らかにした．この結果，本化合物は，臨床適用可能な放射線増感剤の候補物質として有望であることを示した．


Radiosensitizing, Toxicological, and Pharmacokinetic Properties of Hydroxamate Analogues of Nitroimidazoles as Bifunctional Radiosensitizers/Chemical Modifiers

We examined the pharmacokinetics of KIH-802, potassium 2-nitroimidazole-1-acetohydroxamate, using its radioisotope-labelled compound and the acute toxicity in mice. We discovered that the concentration of KIH-802 was very low in the brain and its LD50 was nearly half the value of that of MISO. We also present here new 2-nitroimidazole radiosensitizers/chemical modifiers (KIN-804, 811, 831, 841, 844, 821, 823 and 824) designed to enhance their sensitizing ability intensely by substituting various biologically active groups, such as hydroxamic acids and oximes, with moderate lipophilicity to the aromatic ring, if necessary, through some spacers. The sensitizing effects of all compounds were estimated to be almost equal to or better than that of MISO. The results of their toxicities shows that new hydroxamates KIN-804 and 831 are less toxic than KIH-802 and MISO. Their in vitro enhancement ratios are 2.00 and 1.75, respectively, compared with those of KIH-802, MISO and SR-2508, 1.77, 1.72 and 1.72, respectively, at each dose of 1 mM for EMT6/KU single cell. We concluded that they may be superior radiosensitizers of hydroxamic acid analogues to KIH-802.

本論文において，多機能性低酸素細胞放射線増感剤として，種々のヒドロキサム酸基を有するニトロイミダゾール誘導体を新たに分子設計·合成した．これらはいずれもin vitroで高い活性を有することを明らかにし，特にKIN-804及び831は，毒性が低いことから論文9のKIH-802に比べて優れた放射線増感剤であることを示した．


A Coated Wire Electrode Sensitive to Tetraphenylphosphonium Ion for Measurement of the Mitochondrial Membrane Potential

テトラフェニルホスホニウム(TPP+)イオンに応答する被覆線型電極を開発した．TPP+の10-7∼10-3 Mの濃度範囲でネルンスト応答した．この電極を利用してミトコンドリア膜電位を測定し，ミトコンドリアの呼吸作用の解析に応用した．


KIH-802, An Ace-tohydroxamic Acid Derivative of 2-Nitroimidazole, as A New Potent Hypoxic Cell Ra-diosensitizer: Radiosensitizing Activity, Acute Toxicity, and Pharmacokinetics

The radiosensitizing activity, acute toxicity, and pharmacokinetics of a new hypoxic cell radiosensitizer, potassium 2-nitroimidazole-1-acetohydroxamate (KIH-802), were compared with those of misonidazole (MISO) and etanidazole (SR-2508). The radiosensitizing activity of KIH-802 was slightly higher than that of MISO and SR-2508 in vitro and was similar to or slightly higher than that of MISO or SR-2508 in vivo. The acute toxicity of KIH-802 was slightly higher than that of MISO. The concentrations of KIH-802 in the brains and peripheral nerves of mice were as low as those of SR-2508 and lower than those of MISO.

本論分において，我々が開発した多機能性放射線増感剤KIH-802は，マウスによるin vitro及びin vivo活性試験及び毒性試験の結果，MISOよりも高い活性を有し，低毒性であることが分かった．またMISOと異なり，脳移行率が非常に低いことから神経毒性も改善されるものと予測され，新しい放射線増感剤としての高い有用性を示唆した．


Radiosensitizing Effects of Nitroimidazole Derivatives RP170 and KIH-802


Analysis of Methamphetamine in Hair, Nail, Sweat, and Saliva by Mass Fragmentography

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.


KIH-802: 2-Nitroimidazole-1-acetohydroxamate as a hypoxic Cell Radiosensitizer

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.


Studies on New Triterpenoids of Tripterygium regelii

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.


Isolation of a Hopane-type Trterpenoid, Zeorin, From Higher Plant, Tripterygium regelii


Isolation and Strucuture of New Pseudoquianolides, 4-Epipulchelin and 4-Epineopulchellin From Gaillardia pulchella


Effect of Arylidene-cyclopentenedione Radiosensitizers on ATP Synthesis in Mitochondria:Action as Potent Inhibitors of phosphate Transport

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.

Biochemical and Biophysical Research Communications


Radiosensitizing Effect of the Nitroimidazole Derivatives RK28 and DNIE on Murine Tumors


Glutathione Depletion by 2-Nitroimidazole-1-acetohydroxamic Acid as a Radiosensitizer in Hypoxic Rat Hepatocytes


2-Arylidene-4-cyclopenten-1,3-diones Designed as Non-nitro Radiosensitizers and Hypoxic Cytotoxins

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.


Isolation and Structure of Regelin and Regelinol, New Antitumor Ursene-type Triterpenoids From Tripterygium Regelii

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.


Catalysis by Human Leukocyte Elastase: Proton Inventory as a Mechanistic Probe

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.


Catalysis by Human Leukocyte Elastase: Mechanistic Insights into Specificity Requirements

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.


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)


The Absolute configuration of Regelidine, a Novel 6-Nicotinoyl Dihydroagarofuran Sesquiterpene Alkaloid From Tripterygium Regelii


Radioprotective Phenomena In Murine Tumors Induced by Radiosensitizers at High Dose Levels

放射線増感剤の大量投与時にみられたマウス腫瘍の放射線防護現象


An in vitro and in vivo Screeening System for New Hypoxic Cell Radiosensitizers Using EMT6 Cells


Effect on Hypoxic Cell Radiosensitizers on Glutathione Level and Related Enzyme Activities in Isolated Rat Hepatocytes

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.


A Note on the A-ring Conformation in 2-Chloro-1,2-dihydrosantonins

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.


Chemical Transformation of Aromatin into Pulchellin-A Formal Synthesis of Pulchellidine


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.


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


Studies on Non-sesquiterpenoid Constituents on Gaillarida pulchella. II. Less Lipophilic Substances, Methyl Caffeate as an Antitumor Catecholic


Analysis of Methamphetamine in Human Hair by Mass Fragmentography


Studies on Radioimmunoassay for Methamphetamine Extracted in Human Urine


New Extraction Method of Thromboxane B2 -etermination of TxB2 in Rats with Experimental Endotoxin Shock


Equilibration of 2-Chloro-1,2-dihydrosantonin Conformers; A Theoretical Approach Using X-ray Diffraction and MO Calculations

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.


Cytotoxic Properties of Hydroxylamino- and Amino-misonidazole, Possible Metabolic Products of Misonidazole, in Hypoxic HELA S3 Cells

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.


Structure and Conformation of a New Helenanolide, Pulchelloid C


Hard Acid and Soft Nucleophile System. VI. A Convenient Synthesis of Alkylthiopolycyclic Aromatics with A Metal Halide and Thiol System

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.


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


A Comparative Study of the Radiosensitizing Effects and Cytotoxic Properties of Misonidazole and 2,4-Dinitroimidazole-1-ethanol in Hela S3 Cells

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.


New Tricyclic Ketones Derived from Eudesmanolide


Effect of Solcoseryl® (SS-094) on Wound Healing of Dermal Burns in Ginea Pig (Based on Evaluation of Tissue Collagen)


Radiosensitizing Effect of 2,4-Dinitroimidazole-1-ethanol and its Cytotoxicity in Hela S3 Cells

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.


An Approach to the Design and Synthesis of Mercaptoimidazole Derivatives Containing Oxidized Sulfur based on Electron Affinity Sensitization

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.


An Approach to the Profiling Analysis of Urinary Prostaglandins by Simple Extraction Methods


Structure and Synthesis of Bissantanolides


Simple and Rapid Separation of Certain Prostaglandins by Reversed-phase High-Performance Liquid Chromatography


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


Profiling Analysis of Prostaglandins on Bartter's Syndrome Using a New HPLC Separation Method


Time Course Effect of Anti-inflammatory Drugs on PGF2α and PGE1 as measured by GC/MS


Terpenoids. Part 39. Total Synthesis of Gibberellin A15 and Gibberellin A37

Journal of the Chemical Society, Perkin Transactions 1


Terpenoids. XXXVII. Hypoiodite Reactions with 6-Hydroxy-17-norkaurene- and 7-Norgibberellane-derivatives


Terpenoids. Part XXXVIII. Ring B Contraction of Kaurenolides and Related Compounds into Gibberellane-type Compounds


Demethylation of Aliphatic Methyl Ethers with a Thiol and Boron Trifluoride


Syntheses of Methyl Esters of Gibberellin A15 and Gibberellin A37


システムバイオロジー構築のためのフィチルキノールの合成と抗酸化活性評価

システムバイオロジー構築のためのフィチルキノールの合成と抗酸化活性評価

ビタミンE研究の進歩XIV

ビタミンE研究の進歩XIV


ビタミンEの生合成前駆体およびその非環状類縁体のラジカル消去速度

ビタミンEの生合成前駆体およびその非環状類縁体のラジカル消去速度


イソプレン側鎖を有する植物成分のex vivo 有機合成とその生理活性の評価

イソプレン側鎖を有する植物成分のex vivo 有機合成とその生理活性の評価

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.

Bulletin of Institute of Technology and Science, The University of Tokushima

徳島大学大学院ソシオテクノサイエンス研究部研究報告


ニワトリ卵白のリゾホスホリパーゼDによるリゾホスファチジン酸の産生

ニワトリ卵白のリゾホスホリパーゼDによるリゾホスファチジン酸の産生

Proceedings of the Japanese Conference on the Biochemistry of Lipids


新医薬品の開発をめざした脳神経特異的発現遺伝子産物のプロテオミクス解析

新医薬品の開発をめざした脳神経特異的発現遺伝子産物のプロテオミクス解析

本論文では，新薬開発における新規標的分子の同定のための脳神経特異的遺伝子産物のプロテオミクス解析に関する成果を報告した．

Bulletin of Faculty of Engineering, The University of Tokushima


Introduction of Embryonic Chick Assay on Experiment Applying Nanosecond Pulse Electric Fields on Solid Tumor

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.

IEEJ Transactions on Fundamentals and Materials


Synthesis and photophysical properties of new SNARF derivatives as dual emission pH sensors

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.

Bioorganic & Medicinal Chemistry Letters


Enhanced radical-scavenging activity of naturally-oriented artepillin C derivatives

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.


Suppression of tumor-induced angiogenesis by Brazilian propolis: Major component artepillin C inhibits in vitro tube formation and endothelial cell proliferation

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.