RS6212 is a specific lactate dehydrogenase (LDH) inhibitor (IC50=12.03 μ M）。 RS6212 shows strong anticancer activity in a variety of cancer cell lines.

Most cancer cells switch metabolism from mitochondrial oxidative phosphorylation to aerobic glycolysis, which is catalyzed by lactate dehydrogenase (LDH). Sonic Hedgehog (SHH) pathway aberrant activation is related to metabolism shifting to glycolysis [1]. RS6212 (compound 18) (80 μM; 0-72 h) exhibits significantly anti-proliferative activity against cancer cells and (1 μM, 10 μM, 100 μM; 24 h) inhibits Med-MB (SHH MB, medulloblastoma) with an IC 50 value of 81 μM [1]. RS6212 (80 μM; 6 h) decreases LDH activity, glycolytic level, and ECAR (extracellular acidification rate), and (12.03 μM; 6 h) increases NADH level [1]. RS6212 (0-320 μM; 48 h) inhibits cell growth in HCT116 cells without PARP cleavage nor LC3B–I lipidation [1]. RS6212 (50 nM and 100 nM; 24 h) increases inhibitory effect against HCT116 cells in combination with 50 nM or 100 nM rotenone. RS6212-Rotenone causes significant cleavage of PARP, thus activates programmed cell death of cancer cells [1]. Cell Proliferation Assay [1] Cell Line: HCT116 CRC cells lacking LDHA Concentration: 12.03 μM Incubation Time: 24 hours Result: Failed to inhibit cell proliferation without LDHA, indicating anticancer proliferation by specifically inhibiting LDHA activity. Cell Proliferation Assay [1] Cell Line: Hct116, SW480, A549, PANC-1 Concentration: 80 μM Incubation Time: 0, 24, 48, 72 hours Result: Inhibited cancer cells growth, characterized by increasing glycolytic metabolism.