Tetra-O-methyl-nordihydroguaiaretic acid inhibits energy metabolism and synergistically induces anticancer effects with temozolomide on LN229 glioblastoma tumors - Research article

Sep 24, 2026

Tetra-O-methyl-nordihydroguaiaretic acid (terameprocol; M4N) is evaluated as a transcription inhibitor that, alone and with temozolomide (TMZ), suppresses tumor bioenergetics and enhances anticancer effects. Continuous oral M4N with high-fat diets yields organ exposure in mice and, in LN229 xenografts, inhibits glycolysis and the tricarboxylic acid cycle. Combination therapy decreases LDHA, lactate, NADH/NAD+ salvage regulators and NAD+, reduces oxygen consumption, lowers HIF-1A under hypoxia, and increases stress proteins and reactive oxygen species. In parallel, M4N prevents diet-induced obesity and modulates oncometabolites and itaconate within tumor microenvironments.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages20

Reading Time50 min

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Abstract

Introduction

Results and conclusions (bioenergetics, synergy with temozolomide, in vivo and in vitro effects)

PLOS ONE OPEN ACCESS Citation: Kimura K, Chun JH, Lin Y-L, Liang Y-C, Jackson TLB, Huang RCC (2023) Tetra-O-methylnordihydroguaiaretic acid inhibits energy metabolism and synergistically induces anticancer effects with temozolomide on LN229 glioblastoma tumors implanted in mice while preventing obesity in normal mice that consume high-fat diets. PLoSONE 18(5): e0285536 . [https://doi.org/10.1371/](https://doi.org/10.1371/)[ ](https://doi.org/10.1371/)[journal.pone.0285536](journal.pone.0285536) Editor: Rafael Moreno-Sanchez, Facultad de Estudios Superiores Iztacala, Universidad Nacional Aut´onomade M´exico, MEXICO Received: June 16, 2022 Accepted: April 25, 2023 Published: May 25, 2023 Copyright: © 2023 Kimura et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: The datasets used and/or analyzed during the current and past studies cited in this article are available in a repository on Open Science Framework (OSF), which can be accessed through identifier DOI 10.17605/OSF. IO/ 9M7V8 ([https://doi.org/10.17605/OSF.IO/9M7V8](https://doi.org/10.17605/OSF.IO/9M7V8)) . Any other requests for additional information can RESEARCH ARTICLE Tetra-O-methyl-nordihydroguaiaretic acid inhibits energy metabolism and synergistically induces anticancer effects with temozolomide on LN229 glioblastoma tumors implanted in mice while preventing obesity in normal mice that consume high-fat diets Kotohiko Kimura1, Jong Ho Chun1, Yu-Ling Lin2, Yu-Chuan Liang2, Tiffany L. B. Jackson1, Ru Chih C. Huang1,3 * 1 Department of Biology, Johns Hopkins University, Baltimore, Maryland, United States of America, 2 Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan, Republic of China, 3 Academician, Academia Sinica, Taipei, Taiwan, Republic of China * [rhuang@jhu.edu](rhuang@jhu.edu) Abstract Tetra-O-methyl-nordihydroguaiaretic acid (terameprocol; M4N), a global transcription inhibitor, in combination with a second anticancer drug induces strong tumoricidal activity and has the ability to suppress energy metabolism in cultured cancer cells. In this study, we showed that after continuous oral consumption of high-fat (HF) diets containing M4N, the M4N concentration in most of the organs in mice reached ~1 μM (the M4N concentration in intestines and fat pads was as high as 20–40 μM) and treatment with the combination of M4N with temozolomide (TMZ) suppressed glycolysis and the tricarboxylic acid cycle in LN229 human glioblastoma implanted in xenograft mice. Combination treatment of M4N with TMZ also reduced the levels of lactate dehydrogenase A (LDHA), a key enzyme for glycolysis; lactate, a product of LDHA-mediated enzymatic activity; nicotinamide phosphoribosyltransferase, a rate-limiting enzyme for nicotinamide adenine dinucleotide plus hydrogen (NADH)/NAD+ salvage pathway; and NAD+, a redox electron carrier essential for energy metabolism. It was also shown that M4N suppressed oxygen consumption in cultured LN229 cells, indicating that M4N inhibited oxidative phosphorylation. Treatment with M4Nand TMZ also decreased the level of hypoxia-inducible factor 1A, a major regulator of LDHA, under hypoxic conditions. The ability of M4N to suppress energy metabolism resulted in induction of the stress-related proteins activating transcription factor 4 and cation transport regulator-like protein 1, and an increase in reactive oxygen species production. In addition, the combination treatment of M4N with TMZ reduced the levels of oncometabolites such as 2-hydroxyglutarate as well as the aforementioned lactate. M4N also induced methylidenesuccinic acid (itaconate), a macrophage-specific metabolite with anti-inflammatory activity, in tumor microenvironments. Meanwhile, the ability of M4N to suppress en

FAQs

What is the main purpose of combining M4N with temozolomide (TMZ)?

To test whether M4N synergistically enhances anticancer effects when used together with TMZ, while also suppressing tumor energy metabolism.

How does M4N affect energy metabolism in LN229 glioblastoma?

M4N suppresses glycolysis and the tricarboxylic acid cycle, reduces oxygen consumption consistent with inhibited oxidative phosphorylation, and lowers lactate and related enzymatic and redox components.

Does M4N show effects beyond tumor tissue in normal mice?

Yes. In mice consuming high-fat diets, M4N prevents obesity in normal tissues, indicating a beneficial effect alongside its tumor-targeted metabolic suppression.

Sep 24, 2026

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