Discovery of Highly Potent, Selective, and Brain-Penetrable Leucine-Rich Repeat Kinase 2 (LRRK2) Small Molecule Inhibitors

Estrada, Anthony A.; Liu, Xingrong; Baker-Glenn, Charles; Beresford, Alan; Burdick, Daniel J.; Chambers, M.G.; Chan, Bryan; Chen, Huifen; Ding, Xiao; DiPasquale, Antonio G.; Domínguez, Sara L.; Dotson, Jennafer; Drummond, Jason; Flagella, Michael; Flynn, Sean T.; Fuji, Reina N.; Gill, Andrew B.; Gunzner-Toste, Janet; Harris, S.F.; Heffron, Timothy P.; Kleinheinz, Tracy; Lee, Donna W.; Pichon, Claire Le; Lyssikatos, Joseph P.; Medhurst, Andrew D.; Moffat, John G.; Mukund, Susmith; Nash, Kevin M.; Scearce‐Levie, Kimberly; Sheng, Zejuan; Shore, Daniel G.; Tran, Thuy; Trivedi, Naimisha; Wang, Shumei; Zhang, Shuo; Zhang, Xiaolin; Zhao, Gui‐Ling; Zhu, Haitao; Sweeney, Zachary K.

doi:10.1021/jm301020q

Cited by 130 publications

(133 citation statements)

References 66 publications

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“…The inhibitors were also assayed for inhibition of Janus kinase 2 (JAK2), as previous efforts had confirmed that JAK2/LRRK2 biochemical selectivity correlated with biochemical selectivity against the broader kinome. 22,23 The 3-and 5-amino-linked pyrazoles (2 and 3) demonstrated good JAK2/LRRK2 selectivity but appeared to have inferior potency for LRRK2 in our biochemical screen and cell-based LRRK2 autophosphorlyation assay when compared to the 4-linked aminopyrazoles (5−7). The poor JAK2 selectivity of compound 4 (2.7×) confirmed that a nitrogen lone pair (e.g., compound 2) or 3-/5-substituent was required to achieve good selectivity.…”

Section: P Arkinson's Disease (Pd) Is a Neurodegenerative Disordermentioning

confidence: 87%

Discovery of a Highly Selective, Brain-Penetrant Aminopyrazole LRRK2 Inhibitor

Chan¹,

Estrada²,

Chen³

et al. 2012

ACS Med. Chem. Lett.

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ABSTRACT:The modulation of LRRK2 kinase activity by a selective small molecule inhibitor has been proposed as a potentially viable treatment for Parkinson's disease. By using aminopyrazoles as aniline bioisosteres, we discovered a novel series of LRRK2 inhibitors. Herein, we describe our optimization effort that resulted in the identification of a highly potent, brain-penetrant aminopyrazole LRRK2 inhibitor (18) that addressed the liabilities (e.g., poor solubility and metabolic soft spots) of our previously disclosed anilinoaminopyrimidine inhibitors. In in vivo rodent PKPD studies, 18 demonstrated good brain exposure and engendered significant reduction in brain pLRRK2 levels post-ip administration. The strategies of bioisosteric substitution of aminopyrazoles for anilines and attenuation of CYP1A2 inhibition described herein have potential applications to other drug discovery programs. KEYWORDS: LRRK2, kinase inhibitor, Parkinson's disease, CYP1A2 inhibition P arkinson's disease (PD) is a neurodegenerative disorder that affects approximately 1% of the world's population over the age of 65. 1 Identification of a disease-modifying or neuroprotective therapeutic for PD patients remains a significant challenge. Recently, genetic research has revealed a connection between a missense mutation (G2019S) in the leucine-rich repeat kinase 2 (LRRK2) gene and a number of familial and idiopathic PD cases. 2−9 Significantly, multiple reports have shown that this mutation enhances the kinase activity of LRRK2. 10−17 The modulation of LRRK2 kinase activity by a selective small molecule inhibitor has therefore been proposed as a potential treatment for PD. 18−23 We recently reported the first disclosure of a series of highly potent, selective, and brain-penetrable anilino-pyrimidine LRRK2 inhibitors as exemplified by compound 1. 22,23 Examination of the liabilities of 1 and structurally similar analogues revealed (a) moderate to poor aqueous solubility (thermodynamic solubility of 1 at pH 7.4 < 0.9 μg/mL), (b) potential for ortho-quinoneimine reactive metabolite formation, and (c) a morpholinocarboxamide motif as a major site of metabolism as indicated by metabolite identification studies (MetID). In an effort to improve upon 1 and mitigate potential safety risks, we initiated a campaign to identify a suitable bioisosteric replacement for the aniline functionality. This approach resulted in the discovery of a novel series of highly selective aminopyrazole LRRK2 inhibitors.Docking experiments using a JAK-2-derived homology model of LRRK2 suggested that compound 1 binds in the ATP binding site of LRRK2 as shown in Figure 1. 22,23 In designing anilino-carboxamide replacements, we chose to retain the hinge binding diaminopyrimidine core as well as the C-5 trifluoromethyl group that interacts favorably with the methionine gatekeeper through van der Waal contacts. Additionally, our lead optimization efforts toward 1 established the importance of occupying the vector adjacent to the hingebinding anilino N-H motif (methoxy group in...

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Section: P Arkinson's Disease (Pd) Is a Neurodegenerative Disordermentioning

confidence: 87%

Discovery of a Highly Selective, Brain-Penetrant Aminopyrazole LRRK2 Inhibitor

Chan¹,

Estrada²,

Chen³

et al. 2012

ACS Med. Chem. Lett.

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“…Many kinase inhibitors will block LRRK2 activity, with CZC-25146 and LRRK2-IN1 having high specificity for LRRK2 [157][158][159]. Neither of these kinase inhibitors would be taken into clinical trials owing to their poor blood-brain barrier permeability [159], although more recent inhibitors may be have better distribution in the brain [160][161][162].…”

Section: Therapeutic Strategies For Lrrk2-associated Pdmentioning

confidence: 99%

Heterogeneity of Leucine-Rich Repeat Kinase 2 Mutations: Genetics, Mechanisms and Therapeutic Implications

Rudenko

Cookson

2014

Neurotherapeutics

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Variation within and around the leucine-rich repeat kinase 2 (LRRK2) gene is associated with familial and sporadic Parkinson's disease (PD). Here, we discuss the prevalence of LRRK2 substitutions in different populations and their association with PD, as well as molecular and cellular mechanisms of pathologically relevant LRRK2 mutations. Kinase activation was proposed as a universal molecular mechanism for all pathogenic LRRK2 mutations, but later reports revealed heterogeneity in the effect of mutations on different activities of LRRK2. One mutation (G2019S) increases kinase activity, whereas mutations in the Ras of complex proteins (ROC)-C-terminus of ROC (COR) bidomain impair the GTPase function of LRRK2. Some risk factor variants, including G2385R in the WD40 domain, actually decrease the kinase activity of LRRK2. We suggest a model where LRRK2 mutations exert different molecular mechanisms but interfere with normal cellular function of LRRK2 at different levels of the same downstream pathway. Finally, we discuss the current state of therapeutic approaches for LRRK2-related PD.

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“…The utility of these molecules is, however, largely confined to in vitro assays owing to poor pharmacokinetic qualities and a lack of brain penetration. More recently, two novel LRRK2 kinase inhibitors (GNE-7915 and GNE-0877) were disclosed and used to probe the safety and tolerability profile of LRRK2 kinase inhibition in rodents and nonhuman primates (Estrada et al, 2012;Fuji et al, 2015). However, despite possessing improved biodistribution properties, both compounds inhibit several off-target kinases, and GNE-0877 has been associated with dose limiting toxicity in rats (Fuji et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

MLi-2, a Potent, Selective, and Centrally Active Compound for Exploring the Therapeutic Potential and Safety of LRRK2 Kinase Inhibition

Fell

Mirescu

Basu

et al. 2015

J Pharmacol Exp Ther

250

307

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Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common genetic cause of familial and sporadic Parkinson's disease (PD). That the most prevalent mutation, G2019S, leads to increased kinase activity has led to a concerted effort to identify LRRK2 kinase inhibitors as a potential disease-modifying therapy for PD. An internal medicinal chemistry effort identified several potent and highly selective compounds with favorable drug-like properties. Here, we characterize the pharmacological properties of cis-2,6-dimethyl-4-(6-(5-(1-methylcyclopropoxy)-1H-indazol-3-yl)pyrimidin-4-yl)morpholine (MLi-2), a structurally novel, highly potent, and selective LRRK2 kinase inhibitor with central nervous system activity. MLi-2 exhibits exceptional potency in a purified LRRK2 kinase assay in vitro (IC 50 5 0.76 nM), a cellular assay monitoring dephosphorylation of LRRK2 pSer935 LRRK2 (IC 50 5 1.4 nM), and a radioligand competition binding assay (IC 50 5 3.4 nM). MLi-2 has greater than 295-fold selectivity for over 300 kinases in addition to a diverse panel of receptors and ion channels. Acute oral and subchronic dosing in MLi-2 mice resulted in dosedependent central and peripheral target inhibition over a 24-hour period as measured by dephosphorylation of pSer935 LRRK2. Treatment of MitoPark mice with MLi-2 was well tolerated over a 15-week period at brain and plasma exposures .100Â the in vivo plasma IC 50 for LRRK2 kinase inhibition as measured by pSer935 dephosphorylation. Morphologic changes in the lung, consistent with enlarged type II pneumocytes, were observed in MLi-2-treated MitoPark mice. These data demonstrate the suitability of MLi-2 as a compound to explore LRRK2 biology in cellular and animal models.

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Discovery of Highly Potent, Selective, and Brain-Penetrable Leucine-Rich Repeat Kinase 2 (LRRK2) Small Molecule Inhibitors

Cited by 130 publications

References 66 publications

Discovery of a Highly Selective, Brain-Penetrant Aminopyrazole LRRK2 Inhibitor

Discovery of a Highly Selective, Brain-Penetrant Aminopyrazole LRRK2 Inhibitor

Heterogeneity of Leucine-Rich Repeat Kinase 2 Mutations: Genetics, Mechanisms and Therapeutic Implications

MLi-2, a Potent, Selective, and Centrally Active Compound for Exploring the Therapeutic Potential and Safety of LRRK2 Kinase Inhibition

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