Targeting Nitric Oxide Production in Microglia with Novel Imidazodiazepines for Nonsedative Pain Treatment

Nieman，Amanda N.1; Li，Guanguan2; Zahn，Nicolas M.1; Mian，Md Yeunus1; Mikulsky，Brandon N.3; Hoffman，Dylan A.1; Wilcox，Taylor M.1; Kehoe，Alexander S.1; Luecke，Ian W.1; Poe，Michael M.4; Alvarez-Carbonell，David5; Cook，James M.1; Stafford，Douglas C.1,3; Arnold，Leggy A.1,3

2020-07-01

10.1021/acschemneuro.0c00324

ACS Chemical Neuroscience   影响因子和分区

1948-7193

1948-7193

The goal of this research is the identification of new treatments for neuropathic pain. We characterized the GABAergic system of immortalized mouse and human microglia using electrophysiology and qRT-PCR. Cells from both species exhibited membrane current changes in response to γ-aminobutyric acid, with an EC50 of 260 and 1940 nM, respectively. Human microglia expressed high levels of the γ-aminobutyric acid type A receptor (GABAAR) α3 subunit, which can assemble with β1 and γ2/δ subunits to form functional GABAARs. Mouse microglia contained α2, α3, and α5, in addition to β1-3, γ1-2, and δ, mRNA, enabling a more diverse array of GABAARs than human microglia. Benzodiazepines are well-established modulators of GABAAR activity, prompting a screen of a library of diverse benzodiazepines in microglia for cellular effects. Several active compounds were identified by reduction of nitric oxide (NO) in interferon gamma and lipopolysaccharide activated microglia. However, further investigation with GABAAR antagonists flumazenil, picrotoxin, and bicuculline demonstrated that GABAARs were not linked to the NO response. A screen of 48 receptors identified the κ-opioid receptor and to a lesser extent the μ-opioid receptor as molecular targets, with opioid receptor antagonist norbinaltorphimine reversing benzodiazepine induced reduction of microglial NO. Functional assays identified the downregulation of inducible NO synthase as the mode of action of imidazodiazepines MP-IV-010 and GL-IV-03. Like other κ-opioid receptor agonists, GL-IV-03 reduced the agitation response in both phases of the formalin nociception test. However, unlike other κ-opioid receptor agonists, MP-IV-010 and GL-IV-03 did not impair sensorimotor coordination in mice. Thus, MP-IV-010 and GL-IV-03 represent a new class of nonsedative drug candidates for inflammatory pain.

benzodiazepine formalin test GABAAR microglia nitric oxide opioid receptor

SCI

英语

其他

National Institutes of Health (USA)[R41HL147658][R01NS076517][R01HL118561] ; National Science Foundation, Division of Chemistry[CHE-1625735]

Biochemistry & Molecular Biology ; Pharmacology & Pharmacy ; Neurosciences & Neurology

Biochemistry & Molecular Biology ; Chemistry, Medicinal ; Neurosciences

WOS:000547324200020

AMER CHEMICAL SOC

2-s2.0-85087466986

Scopus

1.Department of Chemistry and Biochemistry and the Milwaukee Institute for Drug Discovery,University of Wisconsin-Milwaukee,Milwaukee,53201,United States
2.Shenzhen Grubbs Institute and Department of Chemistry,Shenzhen Key Laboratory of Small Molecule Drug Discovery and Synthesis,Southern University of Science and Technology,Shenzhen,518055,China
3.
4.Department of Chemistry,Western Michigan University,Kalamazoo,49008,United States
5.Department of Molecular Biology and Microbiology,Case Western Reserve University,Cleveland,44106,United States

Nieman，Amanda N.,Li，Guanguan,Zahn，Nicolas M.,et al. Targeting Nitric Oxide Production in Microglia with Novel Imidazodiazepines for Nonsedative Pain Treatment[J]. ACS Chemical Neuroscience,2020,11(13):2019-2030.

Nieman，Amanda N..,Li，Guanguan.,Zahn，Nicolas M..,Mian，Md Yeunus.,Mikulsky，Brandon N..,...&Arnold，Leggy A..(2020).Targeting Nitric Oxide Production in Microglia with Novel Imidazodiazepines for Nonsedative Pain Treatment.ACS Chemical Neuroscience,11(13),2019-2030.

Nieman，Amanda N.,et al."Targeting Nitric Oxide Production in Microglia with Novel Imidazodiazepines for Nonsedative Pain Treatment".ACS Chemical Neuroscience 11.13(2020):2019-2030.