In the lactate dehydrogenase (LDH) family, Lactate dehydrogenase A (LDHA) is the enzyme that catalyzes the conversion of L-lactate and NAD to pyruvate and NADH in the final step of anaerobic glycolysis. Another isoform, Lactate dehydrogenase B (LDHB) catalyzes the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD+ in a post-glycolysis process. As altered glycolytic metabolism is a feature of cancer cells, inhibition of LDHA and LDHB, thus reducing glycolysis was assumed to have anti-tumor potential. R）-GNE-140 is an LDH inhibitor. Based on enzymatic assays, the inhibitory IC50 values of (R)-GNE-140 against LDHA and LDHB are 3 nM and 5 nM, respectively. Results of a lactate production assay in pancreatic MiaPaca2 cell line shows that (R)-GNE-140 inhibited lactate production with the IC50 of 0.67 μM[2]. In an experiment to determine the IC50s of (R)-GNE-140 across a panel of pancreatic cell lines, it was reported that the 4 cell lines of MiaPaca2, KP2, PSN1 and Hup-T3 are sensitive to (R)-GNE-140 with IC50s below 5 μM. When MiaPaca-2 cells were incubated with 2 μM (R)-GNE-140 for the 1 - 5 days and then assessed for levels of active caspase-3 or sub-2N content, levels of active caspase-3 increased within 1 - 2 d and peaked at around days 2 - 3, while obvious increase in cell death (appearance of cells with a DNA content <2N) was first detected at day 3[3]. In xenograft studies, (R)-GNE-140 was administrated to MiaPaca-2-tumor-bearing mice twice daily at 100, 200, and 400 mg/kg for a total of 7 days. A dose-dependent significant reduction in lactate levels and a corresponding increase in both pyruvate and glycerol-3-phosphate levels was observed within 1h of the last GNE-140 dose but not 6h after the last dose, suggesting rapid clearance of GNE-140 in vivo. Consequently, tumor-bearing mice treated for 21 d with twice daily administration of 100, 200 and 400 mg/kg GNE-140 showed no tumor growth inhibition when compared to vehicle control animals[3].
作用机制
X-ray structure assay revealed that GNE-140 bound to LDHA by forming hydrogen bonds in the active site adjacent to NADH[3].
(R)-GNE-140 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Th17 cells
10 μM
3 days
GNE-140 could decrease the percentage of IL-17+CD4 T cells, suggesting the suppressor role of GNE-140 on Th17 differentiation in Bcl-3-/- mice
Front Immunol. 2022 Sep 9;13:929785.
rat alveolar bone marrow mesenchymal cells (ABMMCs)
10 μM
24 h
Inhibited lactate dehydrogenase A (LDHA) activity, reduced lactate production, thereby suppressing osteogenic differentiation, proliferation, and migration of ABMMCs, and promoting apoptosis
Cells. 2022 Nov 22;11(23):3724.
MDA-MB-231 cells
50 μM
24 h to 7 days
To evaluate the effects of GNE-140 on lactic acid production, glucose utilization, cell viability, and mitochondrial membrane potential in MDA-MB-231 cells. Results showed that GNE-140 fully blocked lactic acid production, inhibited glucose utilization and the glycolytic pathway, but had no adverse effects on cell viability or mitochondrial membrane potential, only attenuating cell division.
Biomolecules. 2021 Nov 24;11(12):1751.
mouse melanoma B16-F10 cells
10 μM
1 h
inhibited LDHA and LDHB activity, mimicking the metabolic phenotype of LDHA/B-DKO cells, characterized by suppressed glycolysis and reactivated OXPHOS
J Biol Chem. 2018 Oct 12;293(41):15947-15961.
human colon adenocarcinoma LS174T
10 μM
1 h
inhibited LDHA and LDHB activity, mimicking the metabolic phenotype of LDHA/B-DKO cells, characterized by suppressed glycolysis and reactivated OXPHOS
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