Pyruvate kinase M2 (PKM2) interacts with phosphotyrosine-containing proteins to regulate glucose metabolism in cells. DASA-58 is a potent activator of recombinant PKM2 with an AC90 value of 680 nM and an AC50 value of 38 nM. In A549-PKM2/kd cell lysates, treatment with 40 μM DASA-58 for 3 hours caused 248% increase of pyruvate kinase activity. Incubation of A549 cells with 0 – 100 μM DASA-58 dose-dependently activated PKM2 with an EC50 value of 19.6 μM. Also in A549 cells, pretreatment with 1 μM DASA-58 prevented pervanadate-induced PKM2 activity inhibition. DASA-58 at 50 μM decreased lactate production in H1299 cells compared to DMSO-treated group. In H1299 cells incubated with [6-14C]-glucose, DASA-58 at 30 μM led to a significant reduction of glucose-derived carbon incorporation into lipids. A549 cells treated with 100 μM DASA-58 showed diminished incorporation of glucose carbons into acetyl-CoA and decreased de novo lipogenesis[3]. Injection of SCID mice with DASA-58-treated PC3 cells (40 μM, 48h incubation) significantly downregulated CAFs-induced lung metastases formation[4].
作用机制
DASA-58 increases PKM2 activity by an allosteric mechanism similar to fructose-1,6-bisphosphate, a glycolytic metabolite fructose that binds to PKM2, inducing a conformational conversion of the enzyme into its most active form[3].
DASA-58 细胞实验
Cell Line
Concentration
Treated Time
Description
References
MDA MB 468
15 µM
72 hours
No significant increase in pyruvate kinase activity
Cancer Metab. 2021 Jan 22;9(1):5.
Bone marrow-derived macrophages (BMDMs)
50 µM
1 hour
DASA-58 effectively reversed the FSTL1o/e-induced increase in glycolysis and reversed FSTL1o/e-induced macrophage M1 polarisation and NF-κB activation.
Gut. 2022 Dec;71(12):2539-2550.
Bone marrow-derived macrophages (BMDMs)
50 µM
1 hour pretreatment followed by 24 hours LPS stimulation
To evaluate the effect of DASA-58 on LPS-induced glycolysis and oxidative phosphorylation. Results showed that DASA-58 significantly inhibited LPS-induced glycolysis but had no significant effect on oxidative phosphorylation.
Cell Metab. 2015 Jan 6;21(1):65-80.
RAW264.7 cells
20 µM
16 hours
DASA-58 and NAC weakened the inhibition of IRD on the aerobic glycolysis while CUT129 and RO8191 partly reduced it.
J Inflamm Res. 2021 Feb 5;14:341-354.
Peritoneal macrophages
50 µM
24 hours
To evaluate the effect of DASA-58 on peritoneal macrophages, results showed similar effects as in BMDMs, with increased FASN and CD36 protein expression.
J Lipid Res. 2020 Mar;61(3):351-364.
Bone marrow-derived macrophages (BMDMs)
50 µM
24 hours
To evaluate the effect of DASA-58 on PKM2 activation, results showed that DASA-58 significantly downregulated LXR-α, ABCA1, and ABCG1 protein expression and augmented FASN and CD36 protein expression.
J Lipid Res. 2020 Mar;61(3):351-364.
K562 cells
40 µM
24 hours
DASA-58 significantly reversed the cytotoxicity of DHA on K562 cells
Drug Des Devel Ther. 2020 May 27;14:2091-2100.
Gastric cancer organoids
50 µM
3 hours
Size-exclusion chromatography analysis showed that DASA-58 treatment resulted in a shift of PKM2 protein into a more tetrameric configuration, especially in NTC organoids.
Proc Natl Acad Sci U S A. 2022 Oct 4;119(40):e2123231119.
MCF7
15 µM
72 hours
Increased pyruvate kinase activity without affecting cell survival
Cancer Metab. 2021 Jan 22;9(1):5.
T47-D
15 µM
72 hours
Increased pyruvate kinase activity without affecting cell survival
Cancer Metab. 2021 Jan 22;9(1):5.
HCC 1443
15 µM
72 hours
Increased pyruvate kinase activity without affecting cell survival
Cancer Metab. 2021 Jan 22;9(1):5.
Mouse primary astrocytes
50 µM
30 min pretreatment followed by 12 hours stimulation
Inhibited PKM2 nuclear translocation, reduced astrocyte glycolytic activity and proliferation
Elife. 2024 Sep 12;13:RP98181.
Peritoneal cells (PECs)
50 µM
30 minutes pretreatment followed by 24 hours LPS stimulation
To evaluate the effect of DASA-58 on LPS-induced pro-IL-1β expression. Results showed that DASA-58 significantly inhibited LPS-induced pro-IL-1β expression.
Cell Metab. 2015 Jan 6;21(1):65-80.
CD4+ T cells
25 µM
48 hours
To evaluate the effect of DASA-58 on TH17 cell differentiation, results showed that DASA-58 inhibited IL-17A production but promoted GM-CSF production.
Sci Signal. 2020 Oct 27;13(655):eaay9217.
HNSCC cell lines (SCC-9, FaDu, Detroit 562, HN, BHY, SCC-154)
30 µM
5 hours or 16 hours
To evaluate the effect of DASA-58 on the glycolytic rate of HNSCC cells. Results showed that DASA-58 increased basal and compensatory glycolysis in some cell lines (e.g., BHY, SCC-154) and decreased the oxygen consumption rate.
Int J Mol Sci. 2022 Jan 11;23(2):775.
MDA MB 231
15 µM
72 hours
Increased pyruvate kinase activity without affecting cell survival
Cancer Metab. 2021 Jan 22;9(1):5.
DASA-58 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
ICR mice
LPS-induced acute lung injury model
Oral
20–80 mg/kg
Once daily for 5 days
Inhibited LPS-induced acute lung injury and inflammatory response
J Inflamm Res. 2021 Feb 5;14:341-354.
Mice
LPS-induced sepsis model and Salmonella typhimurium infection model
Intraperitoneal injection
50mg/kg
Single dose, observed for 2 hours or 24 hours
To evaluate the effect of DASA-58 in vivo on LPS and Salmonella typhimurium-induced inflammatory responses. Results showed that DASA-58 significantly inhibited LPS-induced IL-1β production while boosting IL-10 production, leading to impaired bacterial clearance.
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