Benserazide is a novel inhibitor targeting PKM2 for melanoma treatment

Abstract

The M2 splice isoform of pyruvate kinase (PKM2) is a key enzyme for generating pyruvate and ATP in the glycolytic pathway, whereas the role of PKM2 in tumorigenesis remains a subject of debate. In our study, we found PKM2 is highly expressed in melanoma patients and the malignance is positively correlated with high PKM2 activity and glycolytic capability in melanoma cells. Suppression of PKM2 expression by knocking down markedly attenuated malignant phenotype both in vitro and in vivo, and restoration of PKM2 expression in PKM2 depleted cells could rescue melanoma cells proliferation, invasion and metastasis. With the data indicating PKM2 as a potential therapeutic target, we performed screening for PKM2 inhibitors and identified benserazide (Ben), a drug currently in clinical use. We demonstrated that Ben directly binds to and blocks PKM2 enzyme activity, leading to inhibition of aerobic glycolysis concurrent up-regulation of OXPHOS. Of note, despite PKM2 is very similar to PKM1, Ben does not affect PKM1 enzyme activity. We showed that Ben significantly inhibits cell proliferation, colony formation, invasion and migration in vitro and in vivo. The specificity of Ben was demonstrated by the findings that, suppression of PKM2 expression diminishes the efficacy of Ben in inhibition of melanoma cell growth; ectopic PKM2 expression in normal cells sensitizes cells to Ben treatment. Interestingly, PKM2 activity and aerobic glycolysis are upregulated in BRAFi-resistant melanoma cells. As a result, BRAFi-resistant cells exhibit heightened sensitivity to suppression of PKM2 expression or treatment with Ben both in vitro and in vivo.Y.Z. and Z.H. contributed equally to this work Additional Supporting Information may be found in the online version of this article.

<p>Simultaneous Inhibition of Ornithine Decarboxylase 1 and Pyruvate Kinase M2 Exerts Synergistic Effects Against Hepatocellular Carcinoma Cells</p>

“…5 Targeting PKM2 inhibits the development of HCC by suppressing glycolysis. 6 Several drugs that target PKM2 have shown potential antitumor activity, such as benserazide 7 and compound 3k. 8 Compound 3k showed anti-proliferative activity against colorectal and cervical cancer cells.…”

Phosphorylation regulates cullin-based ubiquitination in tumorigenesis

“… Substrate Kinase Inhibitor/activator Cancer Ref. SPOP CDK4/6 Palbociclib Colon cancer 24 c-Fos EGF Dihydrocapsaicin Neoplastic cell transformation 89 c-Fos EGF R1881 Prostate cancer 90 PYGO2 AKT TCN Various cancers 45 BCL2 PKM2 PKM2 389–405 peptide Glioma 91 BCL2 PKM2 Benserazide Melanoma 92 BCL2 PKM2 Shikonin Various cancers 93 ERG CKI Etoposide Prostate cancer 39 FBW7 ERK SCH772984 Liver cancer 94 P62 PKC δ Rottlerin Hepatocellular carcinomas 97 P62 PKC δ Sotrastaurin Non-small cell lung cancer 98 …”

Adaptive redox homeostasis in cutaneous melanoma

“…Melanoma cells overexpress a redox-depending enzyme PKM2, an isoform of the pyruvate kinase, that converts phosphoenolpyruvate into pyruvate, is the last irreversible reaction of aerobic glycolysis [ 80 ]. ROS oxidizes a specific cysteine residue in PKM2, thus diverting glucose away from lactate production and towards the oxidative branch of PPP leading to increased NADPH production and thus redox homeostasis.…”

“…Melanoma cell invasion and metastasis levels were positively correlated with high PKM2 activity as well as the glycolytic capability. In addition, knockdown of PKM2 markedly attenuated the malignant phenotypes of melanoma cells including cell proliferation, invasion and metastasis in vitro and in vivo , suggesting that PKM2 is a potential therapeutic target in melanoma [ 80 ].…”