Deuterium-Stabilized (R)-Pioglitazone, PXL065, for Treatment of X-Linked Adrenoleukodystrophy (ALD)

Background and aims: X-linked Adrenoleukodystrophy (ALD) is a rare neurometabolic disorder caused by ABCD1- gene mutations, leading to Very-Long-Chain Fatty Acids (VLCFA; in particular C26:0) accumulation, inflammation, mitochondrial impairment and demyelination. PXL065, a clinical-stage deuterium-stabilized(R)-stereoisomer of pioglitazone, retains pioglitazone non-genomic actions but lacks PPARγ activity. As pioglitazone exhibits beneficial effects in ALD models and PXL065 may avoid PPARγ- related side effects, we investigated PXL065 effects of in preclinical models. Methods: Patient-derived fibroblasts and lymphocytes and Abcd1-KO mouse glial cells were exposed to PXL065 (5-10μM) and pioglitazone (10μM) for 7 days. VLCFA content was measured by mass spectrometry, selected gene expression by RT-qPCR, and mitochondrial function using a Seahorse Analyzer (after 72hr). PXL065 or pioglitazone (15mg/kg QD) were administered to 6-8-week or 13-month old Abcd1-KO mice for 8 and 12 weeks, respectively. VLCFA content (mass spectrometry), sciatic nerve axonal morphology (electronic microscopy), and locomotor function (open field test) were measured. Results: In patient and mouse glial cells, PXL065 and pioglitazone corrected C26:0, improved mitochondrial function, increased compensatory Abcd2-3 transporter gene expression, and decreased inflammatory gene expression. In Abcd1-KO mice, C26:0 levels were normalized in plasma and decreased in spinal cord (-55%, p\u3c0.01) and brain (-49%, p\u3c0.0001). Pioglitazone had no effect in spinal cord. Following PXL065 and pioglitazone treatment, abnormal axonal morphology (stellate-shaped cells) was improved but only PXL065 showed significantly improved locomotor test results. Conclusion: Despite reduced PPARγ activity, PXL065 showed substantial signs of efficacy and superior therapeutic potential vs. pioglitazone (in vivo) supporting clinical development for ALD. A Phase 2a study is planned in 2022

Direct AMPK activation corrects NASH in rodents through metabolic effects and direct action on inflammation and fibrogenesis

No approved therapies are available for nonalcoholic steatohepatitis (NASH). Adenosine monophosphate–activated protein kinase (AMPK) is a central regulator of cell metabolism; its activation has been suggested as a therapeutic approach to NASH. Here we aimed to fully characterize the potential for direct AMPK activation in preclinical models and to determine mechanisms that could contribute to efficacy for this disease. A novel small-molecule direct AMPK activator, PXL770, was used. Enzyme activity was measured with recombinant complexes. De novo lipogenesis (DNL) was quantitated in vivo and in mouse and human primary hepatocytes. Metabolic efficacy was assessed in ob/ob and high-fat diet–fed mice. Liver histology, biochemical measures, and immune cell profiling were assessed in diet-induced NASH mice. Direct effects on inflammation and fibrogenesis were assessed using primary mouse and human hepatic stellate cells, mouse adipose tissue explants, and human immune cells. PXL770 directly activated AMPK in vitro and reduced DNL in primary hepatocytes. In rodent models with metabolic syndrome, PXL770 improved glycemia, dyslipidemia, and insulin resistance. In mice with NASH, PXL770 reduced hepatic steatosis, ballooning, inflammation, and fibrogenesis. PXL770 exhibited direct inhibitory effects on pro-inflammatory cytokine production and activation of primary hepatic stellate cells. Conclusion: In rodent models, direct activation of AMPK is sufficient to produce improvements in all core components of NASH and to ameliorate related hyperglycemia, dyslipidemia, and systemic inflammation. Novel properties of direct AMPK activation were also unveiled: improved insulin resistance and direct suppression of inflammation and fibrogenesis. Given effects also documented in human cells (reduced DNL, suppression of inflammation and stellate cell activation), these studies support the potential for direct AMPK activation to effectively treat patients with NASH

Imeglimin prevents human endothelial cell death by inhibiting mitochondrial permeability transition without inhibiting mitochondrial respiration

International audienceImeglimin is the first in a new class of oral glucose-lowering agents, having recently completed its phase 2b trial. As Imeglimin did show a full prevention of β-cell apoptosis, and since angiopathy represents a major complication of diabetes, we studied Imeglimin protective effects on hyperglycemia-induced death of human endothelial cells (HMEC-1). These cells were incubated in several oxidative stress environments (exposure to high glucose and oxidizing agent tert-butylhydroperoxide) which led to mitochondrial permeability transition pore (PTP) opening, cytochrome c release and cell death. These events were fully prevented by Imeglimin treatment. This protective effect on cell death occurred without any effect on oxygen consumption rate, on lactate production and on cytosolic redox or phosphate potentials. Imeglimin also dramatically decreased reactive oxygen species production, inhibiting specifically reverse electron transfer through complex I. We conclude that Imeglimin prevents hyperglycemia-induced cell death in HMEC-1 through inhibition of PTP opening without inhibiting mitochondrial respiration nor affecting cellular energy status. Considering the high prevalence of macrovascular and microvascular complications in type 2 diabetic subjects, these results together suggest a potential benefit of Imeglimin in diabetic angiopathy