Optimisation of LRRK2 inhibitors and assessment of functional efficacy in cell-based models of neuroinflammation

Lenka Munoz; Madeline E Kavanagh; Athena F Phoa; Benjamin Heng; Nicolas Dzamko; Ew-Jun Chen; Munikumar Reddy Doddareddy; Gilles J Guillemin; Michael Kassiou

doi:10.1016/j.ejmech.2015.03.003

Optimisation of LRRK2 inhibitors and assessment of functional efficacy in cell-based models of neuroinflammation

Eur J Med Chem. 2015 May 5:95:29-34. doi: 10.1016/j.ejmech.2015.03.003. Epub 2015 Mar 3.

Authors

Lenka Munoz¹, Madeline E Kavanagh², Athena F Phoa¹, Benjamin Heng³, Nicolas Dzamko⁴, Ew-Jun Chen¹, Munikumar Reddy Doddareddy⁵, Gilles J Guillemin³, Michael Kassiou⁶

Affiliations

¹ School of Medical Sciences, University of Sydney, NSW 2006, Australia.
² School of Chemistry, University of Sydney, NSW 2006, Australia.
³ Australian School of Advanced Medicine, Macquarie University, NSW 2109, Australia.
⁴ School of Medical Sciences, University of New South Wales, Sydney, NSW 2052, Australia; Neuroscience Research Australia, Randwick, NSW 2031, Australia.
⁵ Faculty of Health Sciences, University of Sydney, NSW 2006, Australia.
⁶ School of Chemistry, University of Sydney, NSW 2006, Australia; Faculty of Health Sciences, University of Sydney, NSW 2006, Australia. Electronic address: michael.kassiou@sydney.edu.au.

PMID: 25791676
DOI: 10.1016/j.ejmech.2015.03.003

Abstract

LRRK2IN1 is a highly potent inhibitor of leucine-rich repeat kinase 2 (LRRK2, IC50 = 7.9 nM), an established target for treatment of Parkinson's disease. Two LRRK2IN1 analogues 1 and 2 were synthesised which retained LRRK2 inhibitory activity (1: IC50 = 72 nM; 2: IC50 = 51 nM), were predicted to have improved bioavailability and were efficacious in cell-based models of neuroinflammation. Analogue 1 inhibited IL-6 secretion from LPS-stimulated primary human microglia with EC50 = 4.26 μM. In order to further optimize the molecular properties of LRRK2IN1, a library of truncated analogues was designed based on docking studies. Despite lacking LRRK2 inhibitory activity, these compounds show anti-neuroinflammatory efficacy at micromolar concentration. The compounds developed were valuable tools in establishing a cell-based assay for assessing anti-neuroinflammatory efficacy of LRRK2 inhibitors. Herein, we present data that IL-1β stimulated U87 glioma cell line is a reliable model for neuroinflammation, as data obtained in this model were consistent with results obtained using primary human microglia and astrocytes.

Keywords: LRRK2 inhibitors; Neuroinflammation; Structure–activity relationships.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Anti-Inflammatory Agents / chemistry
Anti-Inflammatory Agents / pharmacology*
Benzodiazepinones / chemistry
Benzodiazepinones / pharmacology*
Cells, Cultured
Glioma / drug therapy*
Glioma / enzymology
Glioma / pathology
Humans
Inflammation / drug therapy*
Inflammation / enzymology
Inflammation / pathology
Interleukin-1beta / pharmacology
Interleukin-6 / metabolism
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
Microglia / cytology
Microglia / drug effects*
Microglia / enzymology
Models, Biological
Protein Kinase Inhibitors / pharmacology*
Protein Serine-Threonine Kinases / antagonists & inhibitors*
Pyrimidines / chemistry
Pyrimidines / pharmacology*

Substances

Anti-Inflammatory Agents
Benzodiazepinones
Interleukin-1beta
Interleukin-6
LRRK2-IN1
Protein Kinase Inhibitors
Pyrimidines
LRRK2 protein, human
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
Protein Serine-Threonine Kinases