Papers - OHE Tomoyuki

Development of a fluorescent-labeled trapping reagent to evaluate the risk posed by acyl-CoA conjugates

Shibazaki C., Mashino T., Ohe T.

Drug Metabolism and Pharmacokinetics  52   100509 2023.10

Research paper (scientific journal), Joint Work, Last author, Corresponding author, Accepted

Fullerene derivatives as inhibitors of the SARS-CoV-2 main protease

Katagishi D., Yasuda D., Takahashi K., Nakamura S., Mashino T., Ohe T.

Bioorganic and Medicinal Chemistry Letters 80   129121 2023.01

Research paper (scientific journal), Joint Work, Last author, Corresponding author, Accepted

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Development of p62-Keap1 protein–protein interaction inhibitors as doxorubicin-sensitizers against non-small cell lung cancer

Daisuke Yasuda, Ippei Yoshida, Riyo Imamura, Daiki Katagishi, Kyoko Takahashi, Hirotatsu Kojima, Takayoshi Okabe, Yoshinobu Ichimura, Masaaki Komatsu, Tadahiko Mashino, Tomoyuki Ohe

Results in Chemistry 4   100609 2022.10

Research paper (scientific journal), Joint Work, Last author, Accepted

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Formation of Reactive Metabolites of Benzbromarone in Humanized-Liver Mice

Cho N., Suemizu H., Kamimura H., Ohe T., Ito F., Akita H., Kobayashi K.

Drug Metabolism and Pharmacokinetics  47   100467 2022.08

Research paper (scientific journal), Accepted

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Synthesis and evaluation of tofacitinib analogs designed to mitigate metabolic activation

Tateishi Y., Shibazaki C., Takahashi K., Nakamura S., Kazuki Y., Mashino T., Ohe

Drug Metabolism and Pharmacokinetics 43   100439 2022.04

Research paper (scientific journal), Joint Work, Accepted

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A bis-pyridinium fullerene derivative significantly induces apoptosis through the generation of ROS in BCR-ABL-positive leukemia cells

Sumi, K., Tago, K., Nakazawa, Y., Takahashi, K., Ohe, T., Mashino, T., Funakoshi-Tago, M.

Eur J Pharmacol 916 ( 5 ) 174714 2022.02

Research paper (scientific journal), Joint Work, Accepted

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Novel mechanism by a bis-pyridinium fullerene derivative to induce apoptosis by enhancing the MEK-ERK pathway in a ROS-independent manner in BCR-ABL-positive CML-derived K562 cells

Sumi, K., Tago, K., Nakazawa, Y., Takahashi, K., Ohe, T., Mashino, T., Funakoshi-Tago, M.

Int J Mol Sci 23 ( 2 ) 749 2022.01

Research paper (scientific journal), Joint Work, Accepted

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A pyridinium‑type fullerene derivative suppresses primary effusion lymphoma cell viability via the downregulation of the Wnt signaling pathway through the destabilization of β‑catenin

Kadota A., Moriguchi M., Watanabe T., Sekine Y., Nakamura S., Yasuno T., Ohe T., Mashino T., Fujimuro M.

Oncology Reports 47   46 2022.01

Research paper (scientific journal), Joint Work, Accepted

Development of a Fluorescent-Labeled Trapping Reagent to Detect Reactive Acyl Glucuronides

Shibazaki, C., Mashita O.,Takahashi K., Nakamura, S., Mashino T., Ohe T.

Chemical Research in Toxicology 34 ( 11 ) 2343 - 2352 2021.11

Research paper (scientific journal), Joint Work, Accepted

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Novel pyridinium-type fullerene derivatives as multitargeting inhibitors of HIV-1 reverse transcriptase, HIV-1 protease, and HCV NS5B polymerase

Kobayashi T, Yasuno T, Takahashi K, Nakamura S, Mashino T, Ohe T

Bioorganic & Medicinal Chemistry Letters 49   128267 2021.10

Research paper (scientific journal), Joint Work, Accepted

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Mitochondrial reactive oxygen species trigger metformin-dependent antitumor immunity via activation of Nrf2/mTORC1/p62 axis in tumor infiltrating CD8T lymphocytes

Nishida M, Yamashita N, Ogawa T, Koseki K, Warabi E, Ohe T, Komatsu M, Kawakami E, Shiroguchi K, Udono H

J Immunotherapy of Cancer  9 ( 9 ) e002954 2021.10

Research paper (scientific journal), Joint Work, Accepted

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Development of Fluorescent-Labeled Trapping Reagents Based on Cysteine to Detect Soft and Hard Electrophilic Reactive Metabolites

Shibazaki C., Ohe T,. Takahashi K., Nakamura S., Mashino T.

Drug Metabolism and Pharmacokinetics 39   100386 2021.08

Research paper (scientific journal), Joint Work, Accepted

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p62/SQSTM1-droplet serves as a platform for autophagosome formation and anti-oxidative stress response

Kageyama, S., Gudmundsson, S., Sou, Y., Ichimura, Y.,, Tamura, N., Kazuno, S., Ueno, T., Miura, Y.,Noshiro, D., Abe, M., Mizushima, T., Miura, N., Okuda, S., Motohashi, H., Lee, J-A., Sakimura, K., Ohe, T., Noda, N., Waguri, S., Eskelinen, E-L., Komatsu, M.

Nature Communications (Nature Communications)  12 ( 1 ) 16 2021.01

Research paper (scientific journal), Joint Work, Accepted

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© 2021, The Author(s). Autophagy contributes to the selective degradation of liquid droplets, including the P-Granule, Ape1-complex and p62/SQSTM1-body, although the molecular mechanisms and physiological relevance of selective degradation remain unclear. In this report, we describe the properties of endogenous p62-bodies, the effect of autophagosome biogenesis on these bodies, and the in vivo significance of their turnover. p62-bodies are low-liquidity gels containing ubiquitin and core autophagy-related proteins. Multiple autophagosomes form on the p62-gels, and the interaction of autophagosome-localizing Atg8-proteins with p62 directs autophagosome formation toward the p62-gel. Keap1 also reversibly translocates to the p62-gels in a p62-binding dependent fashion to activate the transcription factor Nrf2. Mice deficient for Atg8-interaction-dependent selective autophagy show that impaired turnover of p62-gels leads to Nrf2 hyperactivation in vivo. These results indicate that p62-gels are not simple substrates for autophagy but serve as platforms for both autophagosome formation and anti-oxidative stress.

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Fullerene derivatives as dual inhibitors of HIV-1 reverse transcriptase and protease

Yasuno, T., Ohe, T., Kataoka, H., Hashimoto, K., Ishikawa, Y., Furukawa, K., Tateishi, Y., Kobayashi, T., Takahashi, K., Nakamura, S., Mashino, T,

Bioorganic & Medicinal Chemistry Letters (Bioorganic and Medicinal Chemistry Letters)  31   127675 2021.01

Research paper (scientific journal), Joint Work, Accepted,  ISSN  0960894X

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© 2020 Elsevier Ltd In the present study, we newly synthesized three types of novel fullerene derivatives: pyridinium-type derivatives trans-3a and 4a-5b, piperidinium-type derivative 9, and proline-type derivatives 10a-12. Among the assessed compounds, 5a, 10e, 10f, 10i, 11a-d, and 12 were found to inhibit both HIV reverse transcriptase and HIV protease (HIV-PR), with IC50 values in the low micromolar range being observed. Regarding HIV-PR inhibition activity, proline-type derivatives 11a-11d and 12, bearing an alkyl chain between the hydroxylmethylcarbonyl (HMC) moiety and pyrrolidine ring, were more potent than other derivatives. This result might indicate that connecting HMC moieties with proline-type fullerene derivatives through properly sized alkyl chain leads to improved HIV-PR inhibitory activity.

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Development of Novel Diclofenac Analogs Designed to Avoid Metabolic Activation and Hepatocyte Toxicity

Tateishi, Y., Ohe, T., Ogawa, M., Takahashi, K., Nakamura, S., Mashino, T.

ACS Omega (ACS Omega)  5 ( 50 ) 32608 - 32616 2020.12

Research paper (scientific journal), Joint Work, Accepted

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© Diclofenac (DCF) is widely used as a nonsteroidal anti-inflammatory drug; however, it is associated with severe liver injury. This adverse reaction is thought to be related to the reactive quinone imine (QI) and acyl glucuronide (AG) metabolites of DCF, but it remains controversial which reactive metabolites mainly contribute to DCF-induced toxicity. In this study, we synthesized five types of DCF analogs that were designed to mitigate the formation of reactive QI and/or AG metabolites and evaluated their metabolic stability, cyclooxygenase (COX) inhibitory activity, and toxicity to cryopreserved human hepatocytes. Compounds with fluorine at the 5- A nd 4-positions of aromatic rings exhibited modest and high metabolic stability to oxidation by cytochrome P450, respectively, but induced cytotoxicity comparable to DCF. Replacing the carboxylic group of DCF with its bioisosteres was effective in terms of stability to oxidative metabolism and glucuronidation; however, sulfonic acid and sulfonamide groups were not preferable for COX inhibition, and tetrazole-containing analogs induced strong cytotoxicity. On the other hand, compounds that have fluorine at the benzylic position were resistant to glucuronidation and showed little toxicity to hepatocytes. In addition, among these compounds, those with hydrogen at the 4-position (2a and 2c) selectively inhibited the COX-2 enzyme. Throughout these data, it was suggested that compounds 2a and 2c might be novel safer and more efficacious drug candidates instead of DCF.

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Inhibitors of the protein–protein interaction between phosphorylated p62 and Keap1 attenuate chemoresistance in a human hepatocellular carcinoma cell line

Yasuda D., Ohe T., Takahashi K., Imamura R., Kojima H., Okabe T., Ichimura Y., Komatsu M., Yamamoto M., Nagano T., Mashino T.

Free Radical Research 54 ( 11-12 ) 859 - 871 2020.03

Research paper (scientific journal), Accepted,  ISSN  10715762

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© 2020, © 2020 Informa UK Limited, trading as Taylor & Francis Group. Resistance to anticancer agents has been an obstacle to developing therapeutics and reducing medical costs. Whereas sorafenib is used for the treatment of human hepatocellular carcinoma (HCC), resistance limits its efficacy. p62, a multifunctional protein, is overexpressed in several HCC cell lines, such as Huh-1 cells. Phosphorylated p62 (p-p62) inhibits the protein–protein interaction (PPI) between Keap1 and Nrf2, resulting in the Nrf2 overactivation that causes drug resistance. We have found a unique Nrf2 inactivator, named K67, that inhibited the PPI between Keap1 and p-p62 and attenuated sorafenib resistance in Huh-1 cells. Herein, we designed and synthesised novel K67 derivatives by modification of the substituent at the 4-position of the two benzenesulfonyl groups of K67. Although these new derivatives inhibited the Keap1-p-p62 PPI to a level comparable to or weaker than that of K67, the isopropoxy derivative enhanced the sensitivity of Huh-1 cells to sorafenib to a greater extent than K67 without any influence on the viability of Huh-7 cells, which is a non-resistant HCC cell line. The isopropoxy derivative also increased the sensitivity of Huh-1 cells to regorafenib, which suggests that this derivative has the potential to be used as an agent to overcome chemoresistance based on Nrf2 inactivation.

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Synthesis and Evaluation of Nevirapine Analogs to Study the Metabolic Activation of Nevirapine

Yasuhiro Tateishi, Tomoyuki Ohe, Daisuke Yasuda, Kyoko Takahashi, Shigeo Nakamura, Yasuhiro Kazuki, Tadahiko Mashino

Drug Metabolism and Pharmacokinetics (Drug Metabolism and Pharmacokinetics)  35 ( 2 ) 238 - 243 2020.02

Research paper (scientific journal), Joint Work, Accepted,  ISSN  13474367

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Nevirapine (NVP) is widely used as a non-nucleoside reverse transcriptase inhibitor of HIV-1, however, it is associated with severe skin and liver injury. The mechanisms of these adverse reactions are not yet clear, but the metabolic activation of NVP is thought to be related to the injury process. Until now, several metabolic activation pathways of NVP have been reported. In this study, in order to identify the reactive metabolite of NVP mainly responsible for CYP inhibition and liver injury, we synthesized five NVP analogs designed to avoid the proposed bioactivation pathway and evaluated their metabolic stabilities, CYP3A4 time-dependent inhibitory activities, and cytotoxicity. As a result, only a pyrimidine analog of NVP, which could avoid the formation of a reactive epoxide intermediate, did not inhibit CYP3A4. Outside of this compound, the other synthesized compounds, which could avoid the generation of a reactive quinone-methide intermediate, inhibited CYP3A4 equal to or stronger than NVP. The pyrimidine analog of NVP did not induce cytotoxicity in HepG2 and transchromosomic HepG2 cells, expressing major four CYP enzymes and CYP oxidoreductase. These results indicated that the epoxide intermediate of NVP might play an important role in NVP-induced liver injury.

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Synthesis and antitumor activity of novel pyridinium fullerene derivatives

Takumi Yasuno, Tomoyuki Ohe, Hitomi Ikeda, Kyoko Takahashi, Shigeo Nakamura, Tadahiko Mashino

International Journal of Nanomedicine 14   6325 - 6337 2019.08

Research paper (scientific journal), Joint Work, Accepted,  ISSN  11769114

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Purpose: We have previously reported that some cationic fullerene derivatives exhibited anticancer activity, and they are expected to be a potential lead compound for an anti-drug resistant cancer agent. However, they are bis-adducts and a mixture of multiple regioisomers, which cannot be readily separated due to the variability of substituent positions on the fullerene cage. To overcome this issue, we evaluated the antiproliferative activities of a set of mono-adduct derivatives and examined their structure-activity relationship. In addition, the in vivo antitumor activity of selected derivatives was also examined. Methods: Nineteen pyridinium fullerene derivatives were newly designed and synthesized in this study. Their antiproliferative activities were evaluated using several cancer cell lines including drug-resistant cells. Furthermore, in vivo antitumor activity of several derivatives was investigated in mouse xenograft model of human lung cancer. Results: The derivatives inhibited the proliferation of cancer cell lines, including cisplatin-resistant cells and doxorubicin-resistant cells. It was also shown that compound 10 (10 μM), 13 (10 μM) and cis-14 (10 μM) induced the intracellular oxidative stress. In addition, compound 13 (20 mg/kg) and cis-14 (15 mg/kg) significantly exhibited antitumor activity in mouse xenograft model of human lung cancer. Conclusion: We synthesized a novel set of mono-adduct fullerene derivatives functionalized with pyridinium groups and found that most of them show potent antiproliferative activities against cancer cell lines and some of them show significant antitumor activities in vivo. We propose that these fullerene derivatives serve as the lead compounds for a novel type of antitumor agents.

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N-Acetyl cysteine prevents activities of STAT3 inhibitors, Stattic and BP-1-102 independently of its antioxidant properties

Yuki Uchihara, Tomoyuki Ohe, Tadahiko Mashino, Takayuki Kidokoro, Hiroomi Tamura and Megumi Funakoshi-Tago

Pharmacological Reports 71 ( 6 ) 1067 - 1078 2019.05

Research paper (scientific journal), Joint Work, Accepted,  ISSN  17341140

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Inhibitors for signal transducer and activator of transcription 3 (STAT3), Stattic, BP-1-102, and LLL12 significantly induce apoptosis in transformed Ba/F3 cells expressing an oncogenic fusion protein, nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) that induces the activation of STAT3. We found that the antioxidant reagent, N-acetyl cysteine (NAC) prevented the abilities of Stattic and BP-1-102, but not LLL12 to induce apoptosis in transformed cells expressing NPM-ALK, providing a novel problem in use of STAT3 inhibitors. We herein investigated the mechanisms how NAC prevented the effects of Sttatic and BP-1-102. Methods: Ba/F3 cells expressing NPM-ALK and SUDHL-1 cells were treated with antioxidants such as NAC, Trolox or edaravone in combination with STAT3 inhibitors. Phosphorylation of STAT3, cell proliferation rate, cell viability, cell cycle, internucleosomal DNA fragmentation and the intracellular accumulation of reactive oxygen species (ROS) was investigated. The binding of STAT3 inhibitors and NAC was analyzed by LC–MS. Results: NAC but not Trolox and edaravone diminished the abilities of Stattic and BP-1-102 to induce apoptosis in cells expressing NPM-ALK. The ROS levels in cells expressing NPM-ALK were not markedly affected by the treatments with Stattic and BP-1-102 in combination with NAC, suggesting that NAC inhibited the activity of Stattic and BP-1-102 independent of its antioxidant activity. LC–MS analysis revealed that NAC directly bound to Stattic and BP-1-102. Furthermore, these NAC adducts exhibited no cytotoxicity, and failed to affect the activity of STAT3. Conclusions: NAC antagonizes the activities of Stattic and BP-1-102, which inhibit STAT3 activation by interacting with cysteine residues in STAT3.

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Synthesis of novel benzbromarone derivatives designed to avoid metabolic activation

Ohe T, Umezawa R, Kitagawara Y, Yasuda D, Takahashi K, Nakamura S, Abe A, Sekine S, Ito K, Okunushi K, Morio H, Furihata T, Anzai N, Mashino T.

Bioorganic and Medicinal Chemistry Letters 28 ( 23-24 ) 3708 - 3711 2018.10

Research paper (scientific journal), Joint Work, Accepted,  ISSN  0960894X

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We synthesized six novel BBR derivatives that were designed to avoid metabolic activation via ipso-substitution and evaluated for their degree of toxicity and hURAT1 inhibition. It was found that all of the derivatives demonstrate lower cytotoxicity in mouse hepatocytes and lower levels of metabolic activation than BBR, while maintaining their inhibitory activity toward the uric acid transporter. We propose that these derivatives could serve as effective uricosuric agents that have much better safety profiles than BBR.

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