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http://dx.doi.org/10.14348/molcells.2021.0074

A Novel Pyrazolo[3,4-d]pyrimidine Induces Heme Oxygenase-1 and Exerts Anti-Inflammatory and Neuroprotective Effects  

Lee, Ji Ae (Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine)
Kwon, Young-Won (Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine)
Kim, Hye Ri (Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine)
Shin, Nari (Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine)
Son, Hyo Jin (Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine)
Cheong, Chan Seong (Center for Neuro-Medicine, Brain Science Institute, Korea Institute of Science and Technology)
Kim, Dong Jin (Center for Neuro-Medicine, Brain Science Institute, Korea Institute of Science and Technology)
Hwang, Onyou (Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine)
Publication Information
Molecules and Cells / v.45, no.3, 2022 , pp. 134-147 More about this Journal
Abstract
The anti-oxidant enzyme heme oxygenase-1 (HO-1) is known to exert anti-inflammatory effects. From a library of pyrazolo[3,4-d]pyrimidines, we identified a novel compound KKC080096 that upregulated HO-1 at the mRNA and protein levels in microglial BV-2 cells. KKC080096 exhibited anti-inflammatory effects via suppressing nitric oxide, interleukin1β (IL-1β), and iNOS production in lipopolysaccharide (LPS)-challenged cells. It inhibited the phosphorylation of IKK and MAP kinases (p38, JNK, ERK), which trigger inflammatory signaling, and whose activities are inhibited by HO-1. Further, KKC080096 upregulated anti-inflammatory marker (Arg1, YM1, CD206, IL-10, transforming growth factor-β [TGF-β]) expression. In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridinetreated mice, KKC080096 lowered microglial activation, protected the nigral dopaminergic neurons, and nigral damage-associated motor deficits. Next, we elucidated the mechanisms by which KKC080096 upregulated HO-1. KKC080096 induced the phosphorylation of AMPK and its known upstream kinases LKB1 and CaMKKbeta, and pharmacological inhibition of AMPK activity reduced the effects of KKC080096 on HO-1 expression and LPS-induced NO generation, suggesting that KKC080096-induced HO-1 upregulation involves LKB1/AMPK and CaMKKbeta/AMPK pathway activation. Further, KKC080096 caused an increase in cellular Nrf2 level, bound to Keap1 (Nrf2 inhibitor protein) with high affinity, and blocked Keap1-Nrf2 interaction. This Nrf2 activation resulted in concurrent induction of HO-1 and other Nrf2-targeted antioxidant enzymes in BV-2 and in dopaminergic CATH.a cells. These results indicate that KKC080096 is a potential therapeutic for oxidative stress-and inflammation-related neurodegenerative disorders such as Parkinson's disease.
Keywords
heme oxygenase-1; neuroinflammation; Parkinson's disease; pyrazolo[3,4-d]pyrimidine;
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