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Linderapyrone analogue LPD-01 as a cancer treatment agent by targeting importin7

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Abstract

The Wnt/β-catenin signaling pathway plays important roles in several cancer cells, including cell proliferation and development. We previously succeeded in synthesizing a small molecule compound inhibiting the Wnt/β-catenin signaling pathway, named LPD-01 (1), and 1 inhibited the growth of human colorectal cancer (HT-29) cells. In this study, we revealed that 1 inhibits the growth of HT-29 cells stronger than that of another human colorectal cancer (SW480) cells. Therefore, we have attempted to identify the target proteins of 1 in HT-29 cells. Firstly, we investigated the effect on the expression levels of the Wnt/β-catenin signaling pathway-related proteins. As a result, 1 inhibited the expression of target proteins of Wnt/β-catenin signaling pathway (c-Myc and Survivin) and their genes, whereas the amount of transcriptional co-activator (β-catenin) was not decreased, suggesting that 1 inhibited the Wnt/β-catenin signaling pathway without affecting β-catenin. Next, we investigated the target proteins of 1 using magnetic FG beads. Chemical pull-down assay combined with mass spectrometry suggested that 1 directly binds to importin7. As expected, 1 inhibited the nuclear translocation of importin7 cargoes such as Smad2 and Smad3 in TGF-β-stimulated HT-29 cells. In addition, the knockdown of importin7 by siRNA reduced the expression of target genes of Wnt/β-catenin signaling pathway. These results suggest that importin7 is one of the target proteins of 1 for inhibition of the Wnt/β-catenin signaling pathway.

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Acknowledgements

This work was supported by JSPS KAKENHI Grant Numbers 20H03397 and 23KJ2068.

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Authors and Affiliations

  1. Laboratory of Public Health, Kyoto Pharmaceutical University, Misasagi, Yamashina-ku, Kyoto, 607-8412, Japan

    Takahiro Kitagawa, Takahiro Matsumoto & Tetsushi Watanabe

  2. Faculty of Pharmaceutical Sciences, Nagasaki International University, Nagasaki, 859-3298, Japan

    Tomoe Ohta & Tatsusada Yoshida

  3. Laboratory of Biochemistry and Molecular Biology, Kyoto Pharmaceutical University, Misasagi, Yamashina-ku, Kyoto, 607-8412, Japan

    Youhei Saito & Yuji Nakayama

  4. Laboratory of Clinical and Translational Physiology, Kyoto Pharmaceutical University, Misasagi, Yamashina-ku, Kyoto, 607-8412, Japan

    Yuki Hadate & Eishi Ashihara

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  1. Takahiro Kitagawa
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Contributions

Conceptualization and project administration, TM; methodology, TK, TM, TO, TY, YS, YN, YH, and EA; investigation, formal analysis, and data curation, TK, TM, TO, and TY; resources and software, TM, TY, YS, YN, and TW; writing–original draft preparation and visualization, TK; writing–review and editing, TM, YS, and TW; supervision, T.M. and T.W.; funding acquisition, TK and TM; All authors have read and agreed to the published version of the manuscript.

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Correspondence to Takahiro Matsumoto or Tetsushi Watanabe.

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Kitagawa, T., Matsumoto, T., Ohta, T. et al. Linderapyrone analogue LPD-01 as a cancer treatment agent by targeting importin7. J Nat Med 78, 370–381 (2024). https://doi.org/10.1007/s11418-023-01774-y

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