Preclinical and clinical evaluation of the LRRK2 inhibitor DNL201 for Parkinson’s disease

Abstract

Mutations in leucine-rich repeat kinase 2 ( LRRK2 ) are the most common genetic risk factors for Parkinson’s disease (PD). Increased LRRK2 kinase activity is thought to impair lysosomal function and may contribute to the pathogenesis of PD. Thus, inhibition of LRRK2 is a potential disease-modifying therapeutic strategy for PD. DNL201 is an investigational, first-in-class, CNS-penetrant, selective, ATP-competitive, small-molecule LRRK2 kinase inhibitor. In preclinical models, DNL201 inhibited LRRK2 kinase activity as evidenced by reduced phosphorylation of both LRRK2 at serine-935 (pS935) and Rab10 at threonine-73 (pT73), a direct substrate of LRRK2. Inhibition of LRRK2 by DNL201 demonstrated improved lysosomal function in cellular models of disease, including primary mouse astrocytes and fibroblasts from patients with Gaucher disease. Chronic administration of DNL201 to cynomolgus macaques at pharmacologically relevant doses was not associated with adverse findings. In phase 1 and phase 1b clinical trials in 122 healthy volunteers and in 28 patients with PD, respectively, DNL201 at single and multiple doses inhibited LRRK2 and was well tolerated at doses demonstrating LRRK2 pathway engagement and alteration of downstream lysosomal biomarkers. Robust cerebrospinal fluid penetration of DNL201 was observed in both healthy volunteers and patients with PD. These data support the hypothesis that LRRK2 inhibition has the potential to correct lysosomal dysfunction in patients with PD at doses that are generally safe and well tolerated, warranting further clinical development of LRRK2 inhibitors as a therapeutic modality for PD.

Doubly Constrained C-terminal of Roc (COR) Domain-Derived Peptides Inhibit Leucine-Rich Repeat Kinase 2 (LRRK2) Dimerization

“…5 Efforts to develop LRRK2 inhibitors have largely focused on small molecule ATP competitive-binding kinase inhibitors such as MLi-2 and DNL201. 6,7 While these small molecule inhibitors are successful at downregulating LRRK2 kinase activity, there was some evidence of toxicities observed that may be limited to particular animal models. 6,8 Protein−protein interactions (PPIs) are a major driving force for the activation of many cellular pathways and associated disease pathologies, making them an attractive target for drug discovery.…”

LRRK2 and Parkinson's disease: from genetics to targeted therapy

“…The discovery that LRRK2 deleterious variants lead to increased kinase activity 9,18 laid the foundations for LRRK2 kinase inhibitors as a potential treatment for LRRK2 ‐PD (Table 3). The main LRRK2 inhibitors currently developed are DNL151 and DNL201, which already passed phase I and Ib clinical trials with most of the participants developing no or mild adverse effects at clinically relevant doses (https://www.denalitherapeutics.com, 2021) 124 . An indirect mechanism proposed to control LRRK2 activity is the inhibition of its GTPase activity, which demonstrated positive outcomes both in vitro and in vivo , with reduced LRRK2 autophosphorylation and neuroinflammation, as well as suppression of neurodegeneration.…”

“…In mice, histopathological abnormalities have been reported in kidneys after LRRK2 inhibition 130 and, in primates, LRRK2 inhibition produced reversible lamellar bodies in the lungs, but they did not lead to any clinical manifestation 131 . In the DNL151/201 phase 1/1b clinical trials no significant alteration in the pulmonary or renal function emerged (https://www.denalitherapeutics.com, 2021) 124 . In addition, loss of function LRRK2 variants reduce LRRK2 protein levels but are not associated with any phenotypic or pathological alteration 132 .…”

“…On top of measuring the effectiveness of the LRRK2 therapy in clinical trials, they could also be employed to stratify patients based on their response and define the most appropriate dose for each subgroup. Proposed biomarkers, already used in DNL151/201 clinical trials, 124 include phosphorylation of LRRK2 substrates, such as Rab10, or LRRK2 auto‐phosphorylation activity, both associated with LRRK2 kinase function and proven to be appropriate biomarkers to measure target engagement 51 . Another possible biomarker, potentially easier to measure on a clinical setting, is the urinary bis(monoacylglycerol)phosphate (BMP), a phospholipid localized on the late endosome and lysosomal membranes, which regulates the activity of the lysosomal hydrolases and is significantly increased in carriers of the LRRK2 p.G2019S variant 124,136 …”

“…51 Another possible biomarker, potentially easier to measure on a clinical setting, is the urinary bis(monoacylglycerol)phosphate (BMP), a phospholipid localized on the late endosome and lysosomal membranes, which regulates the activity of the lysosomal hydrolases and is significantly increased in carriers of the LRRK2 p.G2019S variant. 124,136 Challenges in LRRK2 trial design and recruitment…”