Figure 1 - available via license: Creative Commons Attribution 4.0 International
Content may be subject to copyright.
Niclosamideactivates PINK1 in HeLa cells. A) Chemical structure of niclosamide. B) Niclosamide dose-response analysis. WT and PINK1 knockout (PINK1 KO) HeLac ells transfected with Parkin were stimulated with either a combination of A/O for 3hor with different concentrations (0.2,0 .8, 2, 8, 20 mm)ofn iclosamide (Niclo) for 40 min. Parkin Ser65 phosphorylation (pS65Parkin), Parkin, Full length OPA1 (F/L), Cleaved OPA1, ubiquitylated CISD1 (CISD1-Ub) and CISD1 weredetected by immunoblotting. GAPDH was used as aloading control.
Source publication
Mutations in PINK1, which impair its catalytic kinase activity, are causal for autosomal recessive early onset Parkinson's disease (PD). Various studies have indicated that the activation of PINK1 could be a useful strategy in treating neurodegenerative diseases such as PD. Herein, we show that the anthelmintic drug niclosamide and its analogues ar...
Citations
... Notably, most research relies on neuronal cultures at early maturation stages or exogenously expressed Parkin 85,93-95 . As previ ous studies have shown, Parkin expression increases with neuronal maturation 96 , correlating with synaptic maturation in vitro 43,96,97 and its localization at both presynaptic 98,99 and postsynaptic terminals in vivo 100,101 . Additionally, neurons undergo extensive mitochondrial metabolic reprogramming during differentiation to reach the final stage of oxidative phosphorylation 102,103 and neuronal maturation, for example, highly structured morphology, electrical excitability and functional synapses 103 . ...
... Notably, most research relies on neuronal cultures at early maturation stages or exogenously expressed Parkin 85,93-95 . As previ ous studies have shown, Parkin expression increases with neuronal maturation 96 , correlating with synaptic maturation in vitro 43,96,97 and its localization at both presynaptic 98,99 and postsynaptic terminals in vivo 100,101 . Additionally, neurons undergo extensive mitochondrial metabolic reprogramming during differentiation to reach the final stage of oxidative phosphorylation 102,103 and neuronal maturation, for example, highly structured morphology, electrical excitability and functional synapses 103 . ...
Mitochondrial dysfunction is a hallmark of idiopathic neurodegenerative diseases, including Parkinson disease, amyotrophic lateral sclerosis, Alzheimer disease and Huntington disease. Familial forms of Parkinson disease and amyotrophic lateral sclerosis are often characterized by mutations in genes associated with mitophagy deficits. Therefore, enhancing the mitophagy pathway may represent a novel therapeutic approach to targeting an underlying pathogenic cause of neurodegenerative diseases, with the potential to deliver neuroprotection and disease modification, which is an important unmet need. Accumulating genetic, molecular and preclinical model-based evidence now supports targeting mitophagy in neurodegenerative diseases. Despite clinical development challenges, small-molecule-based approaches for selective mitophagy enhancement - namely, USP30 inhibitors and PINK1 activators - are entering phase I clinical trials for the first time.
... Previous results point to a drug activating PINK1, as PINK1 levels decreased during transplantation, inconsistent with the increasing expression in natural healthy development. We found three confirmed molecules that successfully activate PINK1: N6-Furfuryladenine, a neo-substrate kinetin [17]; MTK458, a small molecule mitigating pUb and α-synuclein build-up [13]; and Niclosamide, an anthelmintic drug that temporarily disrupts the mitochondrial membrane potential [18]. ...
Parkinsons Disease (PD), the second most prevalent neurodegenerative disease, has the key pathological feature of selective degeneration of dopaminergic neurons (DANs). Current cell therapy based remedy of PD centered on DAN transplantation, whether stem cell derived, or from fetal tissues, yet the possible dysfunction underlying in the microenvironment and non-neuronal mechanisms may impede this solution. The unresolved question is if microenvironment impairments and other non-cell-autonomous signaling may affect the healthy, transplanted DANs, triggering survival rate reduction. We hypothesized that non-neuronal mechanisms do exist thus specific drugs should be incorporated along with DANs during transplantation. Here we performed single cell RNA sequencing (scRNA-seq) analysis using Seurat and AI based package Monocle3 to confirm the existence of non-cell-autonomous signaling and revealed PINK1 and MTF2 as the main culprits through the comparison of initial differentiation dataset and long term transplantation dataset of transplanted DANs. Targeting PINK1, we identified the drug MTK458 and validated its effectiveness through AI based drug screening and molecular docking. Together, these findings supported MTK458 to be an effective co-transplant material designated for alleviating the abnormality in non-neuronal mechanisms or microenvironment.
... It is well documented that niclosamide can inhibit different molecular targets (STAT3, mTOR, Wnt/b-catenin, S100A4, SQSTM1/p62, NF-kB, Notch, TMEM16) [9,10] which are all dysregulated and pathogenic in ALS [11][12][13], suggesting its application to interfere with these altered mechanisms to improve multiple aspects of the pathology. Moreover, it has been demonstrated that niclosamide increases motoneuron differentiation and TDP43 clearance [14,15] and exerts neuroprotective effects in peripheral neuropathies [16] and in models of Parkinson's disease [17,18]. ...
... Previous studies in experimental models of neurodegenerative diseases have suggested that niclosamide may possess neuroprotective properties through various mechanisms [43], including promoting autophagy [17,44]. We showed that niclosamide reduces the cytoplasmic accumulation of phosphorylated TDP43 in the motoneurons of SOD1-G93A mice, a critical pathological feature observed in sporadic and familial forms of ALS [45]. ...
Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease influenced by genetic, epigenetic, and environmental factors, resulting in dysfunction in cellular and molecular pathways. The limited efficacy of current treatments highlights the need for combination therapies targeting multiple aspects of the disease. Niclosamide, an anthelminthic drug listed as an essential medicine, has been repurposed in clinical trials for different diseases due to its anti-inflammatory and anti-fibrotic properties. Niclosamide can inhibit various molecular pathways (e.g., STAT3, mTOR) that are dysregulated in ALS, suggesting its potential to disrupt these altered mechanisms associated with the pathology. We administered niclosamide intraperitoneally to two transgenic murine models, SOD1-G93A and FUS mice, mimicking key pathological processes of ALS. The treatment was initiated at the onset of symptoms, and we assessed disease progression by neurological scores, rotarod and wire tests, and monitored survival. Furthermore, we investigated cellular and molecular mechanisms affected by niclosamide in the spinal cord and muscle of ALS mice. In both models, the administration of niclosamide resulted in a slowdown of disease progression, an increase in survival rates, and an improvement in tissue pathology. This was characterised by reduced gliosis, motor neuron loss, muscle atrophy, and inflammatory pathways. Based on these results, our findings demonstrate that niclosamide can impact multiple pathways involved in ALS. This multi-targeted approach leads to a slowdown in the progression of the disease, positioning niclosamide as a promising candidate for repurposing in the treatment of ALS.
... Therefore, we tested the therapeutic potential of different PINK1-kinase activators to rescue mitochondrial turnover and bioenergetics in the context of diabetesinduced hyperfusion. Four chemical entities were selected based on their reported efficacy in activating PINK1-dependent mitophagy; a) two N6-furfuryladenines (kinetin and its glycosylated metabolite kinetin riboside [KR]) 19,20 ; b) salicylanilides (niclosamide) 31,32 ; c) benzocoumarins (urolithin-A) 33,34 . ...
The recovery of mitochondrial quality control (MQC) may bring innovative solutions for neuroprotection, while imposing a significant challenge given the need of holistic approaches to restore mitochondrial dynamics (fusion/fission) and turnover (mitophagy and biogenesis). In diabetic retinopathy, this is compounded by our lack of understanding of human retinal neurodegenera-tion, but also how MQC processes interact during disease progression. Here, we show that mitochondria hyperfusion is characteristic of retinal neurode-generation in human and murine diabetes, blunting the homeostatic turnover of mitochondria and causing metabolic and neuro-inflammatory stress. By mimicking this mitochondrial remodelling in vitro, we ascertain that N6-furfuryladenosine enhances mitochondrial turnover and bioenergetics by relaxing hyperfusion in a controlled fashion. Oral administration of N6-furfuryladenosine enhances mitochondrial turnover in the diabetic mouse retina (Ins2 Akita males), improving clinical correlates and conferring neuro-protection regardless of glycaemic status. Our findings provide translational insights for neuroprotection in the diabetic retina through the holistic recovery of MQC. With the incidence of diabetes rising at an alarming rate (615 million people projected worldwide by 2045) 1 , the development of therapies aimed at preventing its major complications is of paramount importance. Diabetic retinopathy (DR), a leading cause of blindness in the working-age population, is one of the most prevalent complications, affecting~80% of people who have had diabetes for 1-2 decades 2. During the early stages of pathogenesis (non-proliferative DR [NPDR]), microvascular dysfunction and neural retinal degeneration constitute the key pathologies. At the neuronal level, DR is clinically manifested by electrophysiological deficits and visual field (frequency doubling
... Niclosamide, an antihelminthic drug, has been recognized for its ability to modulate mitochondrial phosphorylation and influence various signaling pathways, including mTOR and JAK/STAT3 [89]. Niclosamide has shown promise in activating PINK1, a kinase involved in protecting against mitochondrial dysfunction, which is particularly relevant in autosomal recessive PD [90]. Niclosamide also promotes neurite growth in dopaminergic neurons and protects against α-Syn-induced neurodegeneration through the BMP-Smad pathway [91]. ...
... Targeting PINK1/Parkin dependent and independent pathways has been suggested. For example, PINK1 activator KTP (kinetin triphosphate) and its analogues and niclosamide and its analogues have been described [138][139][140][141]. ...
Parkinson’s disease (PD) is a devastating disease associated with accumulation of α-synuclein (α-Syn) within dopaminergic neurons, leading to neuronal death. PD is characterized by both motor and non-motor clinical symptoms. Several studies indicate that autophagy, an important intracellular degradation pathway, may be involved in different neurodegenerative diseases including PD. The autophagic process mediates the degradation of protein aggregates, damaged and unneeded proteins, and organelles, allowing their clearance, and thereby maintaining cell homeostasis. Impaired autophagy may cause the accumulation of abnormal proteins. Incomplete or impaired autophagy may explain the neurotoxic accumulation of protein aggregates in several neurodegenerative diseases including PD. Indeed, studies have suggested the contribution of impaired autophagy to α-Syn accumulation, the death of dopaminergic neurons, and neuroinflammation. In this review, we summarize the recent literature on the involvement of autophagy in PD pathogenesis.
... In an excellent and comprehensive review, Chen et al. (2018), and recently updated by Wang et al. (2022) for pharmacological activities and therapeutic prospects, report that niclosamide, tested mainly in cells and a few preclinical animal cancer models, has efficacy that includes "cancer, bacterial infection, metabolic diseases such as type II diabetes, NASH and NAFLD, artery constriction, endometriosis, neuropathic pain, rheumatoid arthritis, sclerodermatous graft-versus-host disease, and systemic sclerosis". There are also reports of activity in Parkinson's (Barini et al. 2018) and COPD (Cabrita et al. 2019;Miner et al. 2019). Niclosamide is a career builder should anyone want to study it in detail. ...
Motivation
The low solubility, weak acid drug, niclosamide is a host cell modulator with broad-spectrum anti-viral cell-activity against many viruses, including stopping the SARS-CoV-2 virus from infecting cells in cell culture. As a result, a simple universal nasal spray preventative was proposed and investigated in earlier work regarding the dissolution of niclosamide into simple buffers. However, starting with pharmaceutical grade, niclosamide represents a new 505(b)(2) application. The motivation for this second paper in the series was therefore to explore if and to what extent niclosamide could be extracted from commercially available and regulatory-approved niclosamide oral tablets that could serve as a preventative nasal spray and an early treatment oral/throat spray, with possibly more expeditious testing and regulatory approval.
Experimental
Measurements of supernatant niclosamide concentrations were made by calibrated UV-Vis for the dissolution of niclosamide from commercially available Yomesan crushed into a powder for dissolution into Tris Buffer (TB) solutions. Parameters tested were as follows: time (0–2 days), concentration (300 µM to -1 mM), pH (7.41 to 9.35), and anhydrous/hydrated state. Optical microscopy was used to view the morphologies of the initial crushed powder, and the dissolving and equilibrating undissolved excess particles to detect morphologic changes that might occur.
Results
Concentration dependence : Niclosamide was readily extracted from powdered Yomesan at pH 9.34 TB at starting Yomesan niclosamide equivalents concentrations of 300 µM, 600 µM, and 1 mM. Peak dissolved niclosamide supernatant concentrations of 264 µM, 216 µM, and 172 µM were achieved in 1 h, 1 h, and 3 h respectively. These peaks though were followed by a reduction in supernatant concentration to an average of 112.3 µM ± 28.4 µM after overnight stir on day 2. pH dependence : For nominal pHs of 7.41, 8.35, 8.85, and 9.35, peak niclosamide concentrations were 4 µM, 22.4 µM, 96.2 µM, and 215.8 µM, respectively. Similarly, the day 2 values all reduced to 3 µM, 12.9 µM, 35.1 µM, and 112.3 µM. A heat-treatment to 200 °C dehydrated the niclosamide and showed a high 3 h concentration (262 µM) and the least day-2 reduction (to 229 µM). This indicated that the presence, or formation during exposure to buffer, of lower solubility polymorphs was responsible for the reductions in total solubilities. These morphologic changes were confirmed by optical microscopy that showed initially featureless particulate-aggregates of niclosamide could grow multiple needle-shaped crystals and form needle masses, especially in the presence of Tris-buffered sodium chloride, where new red needles were rapidly made. Scale up : A scaled-up 1 L solution of niclosamide was made achieving 165 µM supernatant niclosamide in 3 h by dissolution of just one fifth (100 mg niclosamide) of a Yomesan tablet.
Conclusion
These comprehensive results provide a guide as to how to utilize commercially available and approved tablets of niclosamide to generate aqueous niclosamide solutions from a simple dissolution protocol. As shown here, just one 4-tablet pack of Yomesan could readily make 165 L of a 20 µM niclosamide solution giving 16,500 10 mL bottles. One million bottles, from just 60 packs of Yomesan, would provide 100 million single spray doses for distribution to mitigate a host of respiratory infections as a universal preventative-nasal and early treatment oral/throat sprays throughout the world.
Graphical Abstract
pH dependence of niclosamide extraction from crushed Yomesan tablet material into Tris buffer (yellow-green in vial) and Tris-buffered saline solution (orange-red in vial). Initial anhydrous dissolution concentration is reduced by overnight stirring to likely monohydrate niclosamide; and is even lower if in TBSS forming new niclosamide sodium needle crystals grown from the original particles.
... Until now, efforts have been taken to screen small molecules 26-28 such as Niclosamide which is an approved anthelmintic drug used in treating tapeworm infections 29 . An alternate strategy was to use neosubstrate kinetin triphosphate which is an ATP analogue, shown to increase the activity of mutant G309D and wildtype. ...
Mutations in PINK1 kinase are known to be responsible for the early-onset of Parkinson's disease. The protein resides on the outer-membrane of healthy mitochondria, and acts as a quality controller by regulating and maintaining dysfunctional mitochondria also known as mitophagy. Limited knowledge of the mechanism of protein has made it cumbersome to elucidate an effective treatment. In this paper, we screen a set of kinase inhibitors using high-throughput screening for investigating potential activators. We have for the first time shown the plausible binding and selectivity towards the kinase inhibitors in the mutant PINK1 binding site. We also could highlight the ligand bind/unbinding pathway identifying the most stable binding pose utilizing the residues from the P-loop and DFG motif for drugs Tepotinib and Pyrotinib. Interestingly, no direct interaction of mutated residues with the ligand molecules were observed. Furthermore, we highlight that the Ins3 region and SER228 of the N-lobe display distinct conformational states that are inline with crystal structure for substrate binding specificity. Collectively, our findings provide a molecular basis for the PINK1 potential kinase activator, which would further facilitate as a starting point to probe new therapeutics by using the structure based drug designing for treating neurological disorder.
... This drug kills tapeworms by inhibiting oxidative phosphorylation in the mitochondria and anaerobic adenosine triphosphate (ATP) production [1]. Recent studies have shown its potential in treating many other diseases including Parkinson's disease, diabetes, viral and microbial infections, and inhibition of myopia progression [2][3][4][5]. In addition, other studies have demonstrated the efficacy of niclosamide in the treatment of various cancers [6]. ...
Background
To report the first case of bull’s eye maculopathy associated with veterinary niclosamide.
Case presentation
A 27-year-old Iranian female presented with a history of reduced vision and photopsia since 3 years, after accidental ingestion of four boluses of veterinary niclosamide. Fundus examination showed atrophy in parafoveal retinal pigmentary epithelium, appearing as bilateral bull’s-eye maculopathy. Optical coherence tomography revealed disruption of the parafoveal ellipsoid zone and outer retinal thinning, appearing as a flying saucer sign. Electroretinography displayed decreased scotopic and photopic amplitudes with normal waveform in both eyes. The causality score was 4, showing “possible” retinopathy due to niclosamide according to Naranjo’s causality assessment algorithm. Based on clinical and ancillary findings, a diagnosis of niclosamide-induced maculopathy was made.
Conclusion
Veterinary niclosamide is an anthelmintic drug that in higher doses could be detrimental to the human retina. Awareness about its side effects and appropriate drug labeling could prevent accidental toxicity.
... Nonetheless, its significant cell toxicity has led to the development of safer molecules such as 19-Ph-GAt [190]. Another promising therapeutic target is PTEN-induced kinase 1 (PINK1), since its mutation is known to cause the early onset of PD [191,192]. Niclosamide, a salicylanilide drug used to treat parasitic infections, promotes PINK1 activation by driving mitochondrial depolarization [191,192]. Alternatively, other approaches involve chloramphenicol treatment. ...
... Another promising therapeutic target is PTEN-induced kinase 1 (PINK1), since its mutation is known to cause the early onset of PD [191,192]. Niclosamide, a salicylanilide drug used to treat parasitic infections, promotes PINK1 activation by driving mitochondrial depolarization [191,192]. Alternatively, other approaches involve chloramphenicol treatment. ...
Neurodegenerative diseases are characterized by the progressive loss of neurons, synapses, dendrites, and myelin in the central and/or peripheral nervous system. Actual therapeutic options for patients are scarce and merely palliative. Although they affect millions of patients worldwide, the molecular mechanisms underlying these conditions remain unclear. Mitochondrial dysfunction is generally found in neurodegenerative diseases and is believed to be involved in the pathomechanisms of these disorders. Therefore, therapies aiming to improve mitochondrial function are promising approaches for neurodegeneration. Although mitochondrial-targeted treatments are limited, new research findings have unraveled the therapeutic potential of several groups of antibiotics. These drugs possess pleiotropic effects beyond their anti-microbial activity, such as anti-inflammatory or mitochondrial enhancer function. In this review, we will discuss the controversial use of antibiotics as potential therapies in neurodegenerative diseases.
