DASA-58

DASA-58 is an selective activator of pyruvate kinase M2 (PKM2) with an AC90 value of 680 nM, and an AC50 value of 38 nM [1].

In cancer, the altered glucose metabolism can be influenced by the regulatory properties of PKM2. The interaction between PKM2 and phosphotyrosine-containing proteins results in the inhibition of enzyme activity and hence more glycolytic metabolites [1].

In A549-PKM1/kd and A549-PKM2/kd cells, Flag-PKM1 and Flag-PKM2 were expressed, respectively. Endogenous PKM2 was knockdowned. Treatment with DMSO resulted in 233±27% more pyruvate kinase activity in lysates from A549-PKM1/kd cells than that in lysates from A549-PKM2/kd cells. Treatment with DASA-58 did not increase the pyruvate kinase activity in A549-PKM1/kd cells, but it resulted in a 248 ±21% pyruvate kinase activity increase in the lysate of A549-PKM2/kd cells. In cells, 0-100 μM of DASA-58 dose-dependently activated PKM2 with an EC50 value of 19.6 μM [1].

TEPP-46 is another PKM2 activator. In A549 xenograft tumors, TEPP-46 at an acute oral-dose of 150 mg/kg resulted in the maximal activation of PKM2. In mice bearing H1299 xenograft tumors, treatment with TEPP-46 at a 5-day repeat-dose of 50 mg/kg twice daily made tumors harbor exclusively tetrameric PKM2. In xenografts from vehicle-treated mice, little tetrameric PKM2 was found [1].

Reference:
[1].  Anastasiou D, Yu Y, Israelsen WJ, et al. Pyruvate kinase M2 activators promote tetramer formation and suppress tumorigenesis. Nature chemical biology, 2012, 8(10): 839-847.

Targeting stromal-induced pyruvate kinase M2 nuclear translocation impairs oxphos and prostate cancer metastatic spread.[Pubmed:26183399]

Oncotarget. 2015 Sep 15;6(27):24061-74.

Cancer associated fibroblasts (CAFs) are key determinants of cancer progression. In prostate carcinoma (PCa), CAFs induce epithelial-mesenchymal transition (EMT) and metabolic reprogramming of PCa cells towards oxidative phosphorylation (OXPHOS), promoting tumor growth and metastatic dissemination. We herein establish a novel role for pyruvate kinase M2 (PKM2), an established effector of Warburg-like glycolytic behavior, in OXPHOS metabolism induced by CAFs. Indeed, CAFs promote PKM2 post-translational modifications, such as cysteine oxidation and Src-dependent tyrosine phosphorylation, allowing nuclear migration of PKM2 and the formation of a trimeric complex with hypoxia inducible factor-1alpha (HIF-1alpha) and the transcriptional repressor Differentially Expressed in Chondrocytes-1 (DEC1). DEC1 recruitment is mandatory for downregulating miR205 expression, thereby fostering EMT execution and metabolic switch toward OXPHOS. Furthermore, the analysis of a cohort of PCa patients reveals a significant positive correlation between PKM2 nuclear localization and cancer aggressiveness, thereby validating our in vitro observations. Crucially, in vitro and in vivo pharmacological targeting of PKM2 nuclear translocation using DASA-58, as well as metformin, impairs metastatic dissemination of PCa cells in SCID mice. Our study indicates that impairing the metabolic tumor:stroma interplay by targeting the PKM2/OXPHOS axis, may be a valuable novel therapeutic approach in aggressive prostate carcinoma.

Pyruvate kinase M2 regulates Hif-1alpha activity and IL-1beta induction and is a critical determinant of the warburg effect in LPS-activated macrophages.[Pubmed:25565206]

Cell Metab. 2015 Jan 6;21(1):65-80.

Macrophages activated by the TLR4 agonist LPS undergo dramatic changes in their metabolic activity. We here show that LPS induces expression of the key metabolic regulator Pyruvate Kinase M2 (PKM2). Activation of PKM2 using two well-characterized small molecules, DASA-58 and TEPP-46, inhibited LPS-induced Hif-1alpha and IL-1beta, as well as the expression of a range of other Hif-1alpha-dependent genes. Activation of PKM2 attenuated an LPS-induced proinflammatory M1 macrophage phenotype while promoting traits typical of an M2 macrophage. We show that LPS-induced PKM2 enters into a complex with Hif-1alpha, which can directly bind to the IL-1beta promoter, an event that is inhibited by activation of PKM2. Both compounds inhibited LPS-induced glycolytic reprogramming and succinate production. Finally, activation of PKM2 by TEPP-46 in vivo inhibited LPS and Salmonella typhimurium-induced IL-1beta production, while boosting production of IL-10. PKM2 is therefore a critical determinant of macrophage activation by LPS, promoting the inflammatory response.