Isolation, Structural Identification, Synthesis, and Pharmacological Profiling of 1,2-<i>trans</i>-Dihydro-1,2-diol Metabolites of the Utrophin Modulator Ezutromid

Chatzopoulou, Maria; Claridge, Timothy D. W.; Davies, Kay E.; Davies, Stephen G.; Elsey, David J.; Emer, Enrico; Fletcher, Ai M.; Harriman, Shawn; Robinson, Neil; Rowley, Jessica; Russell, Angela J.; Tinsley, Jonathon M.; Weaver, Richard J.; Wilkinson, Isabel V.L.; Willis, Nicky J.; Wilson, Francis X.; Wynne, Graham M.

doi:10.1021/acs.jmedchem.9b01547

Cited by 10 publications

(19 citation statements)

References 38 publications

(81 reference statements)

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“…However, follow-up studies point to poor physicochemical properties as the primary reason for Ezutromid's lack of efficacy. Recent analysis of Ezutromid revealed that it was primarily cleared by cytochrome P450 1A (CYP1A) in the liver into less active metabolites [42]. In rats and pigs given multiple doses, Ezutromid increased biomarkers of CYP1A, further increasing its degradation.…”

Section: Discussionmentioning

confidence: 99%

High-throughput screening identifies modulators of sarcospan that stabilize muscle cells and exhibit activity in the mouse model of Duchenne muscular dystrophy

et al. 2020

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Background Duchenne muscular dystrophy (DMD) is a degenerative muscle disease caused by mutations in the dystrophin gene. Loss of dystrophin prevents the formation of a critical connection between the muscle cell membrane and the extracellular matrix. Overexpression of sarcospan (SSPN) in the mouse model of DMD restores the membrane connection and reduces disease severity, making SSPN a promising therapeutic target for pharmacological upregulation. Methods Using a previously described cell-based promoter reporter assay of SSPN gene expression (hSSPN-EGFP), we conducted high-throughput screening on libraries of over 200,000 curated small molecules to identify SSPN modulators. The hits were validated in both hSSPN-EGFP and hSSPN-luciferase reporter cells. Hit selection was conducted on dystrophin-deficient mouse and human myotubes with assessments of (1) SSPN gene expression using quantitative PCR and (2) SSPN protein expression using immunoblotting and an ELISA. A membrane stability assay using osmotic shock was used to validate the functional effects of treatment followed by cell surface biotinylation to label cell surface proteins. Dystrophin-deficient mdx mice were treated with compound, and muscle was subjected to quantitative PCR to assess SSPN gene expression. Results We identified and validated lead compounds that increased SSPN gene and protein expression in dystrophin-deficient mouse and human muscle cells. The lead compound OT-9 increased cell membrane localization of compensatory laminin-binding adhesion complexes and improved membrane stability in DMD myotubes. We demonstrated that the membrane stabilizing benefit is dependent on SSPN. Intramuscular injection of OT-9 in the mouse model of DMD increased SSPN gene expression. Conclusions This study identifies a pharmacological approach to treat DMD and sets the path for the development of SSPN-based therapies.

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Section: Discussionmentioning

confidence: 99%

High-throughput screening identifies modulators of sarcospan that stabilize muscle cells and exhibit activity in the mouse model of Duchenne muscular dystrophy

et al. 2020

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“…Based on these results, Summit Therapeutics abandoned the development program of ezutromid [ 97 , 98 ]. Recent studies have elucidated the ezutromid mechanism of action as an aryl hydrocarbon receptor (AhR) antagonist [ 53 , 99 ]. Similarly, other molecules that ameliorate mdx pathology like SMT022357 [ 53 ] or resveratrol [ 100 ] have also shown activity as AhR antagonists [ 100 ].…”

Section: Indirect Mechanisms For Utrophin Overexpressionmentioning

confidence: 99%

Utrophin modulator drugs as potential therapies for Duchenne and Becker muscular dystrophies

Soblechero-Martín

López-Martínez

Puente-Ovejero

et al. 2021

Neuropathology Appl Neurobio

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Utrophin is an autosomal paralogue of dystrophin, a protein whose deficit causes Duchenne and Becker muscular dystrophies (DMD/BMD). Utrophin is naturally overexpressed at the sarcolemma of mature dystrophin‐deficient fibres in DMD and BMD patients as well as in the mdx Duchenne mouse model. Dystrophin and utrophin can co‐localise in human foetal muscle, in the dystrophin‐competent fibres from DMD/BMD carriers, and revertant fibre clusters in biopsies from DMD patients. These findings suggest that utrophin overexpression could act as a surrogate, compensating for the lack of dystrophin, and, as such, it could be used in combination with dystrophin restoration therapies. Different strategies to overexpress utrophin are currently under investigation. In recent years, many compounds have been reported to modulate utrophin expression efficiently in preclinical studies and ameliorate the dystrophic phenotype in animal models of the disease. In this manuscript, we discuss the current knowledge on utrophin protein and the different mechanisms that modulate its expression in skeletal muscle. We also include a comprehensive review of compounds proposed as utrophin regulators and, as such, potential therapeutic candidates for these muscular dystrophies.

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“…Promising efficacy and evidence of target engagement was observed in DMD patients after 24 weeks of treatment; however, trial endpoints were not met after 48 weeks. These investigators subsequently generated data that explains the lack of sustained efficacy in the trial [29] and elucidated the molecular mechanism of action (MoA) of ezutromid [30].…”

Section: Replacement Of Dystrophin With a Surrogate Proteinmentioning

confidence: 99%

“…The lack of sustained efficacy in the trial may be explained by the extensive metabolism of ezutromid via CYP1A oxidation to predominantly two dihydrodiol metabolites in humans, both of which when tested for utrophin up-regulation showed substantially reduced potency or no activity [29]. Moreover, repeated dosing of ezutromid leads to a reduction in exposure in both healthy volunteers (∼20% reduction after 10 days) and markedly in DMD patients (∼60% reduction after 10 days) [31] through induction of CYP1A [29,32,33]. This means early in the trial, the patients are exposed to sufficient ezutromid to achieve target engagement and utrophin up-regulation, but at later timepoints exposure is insufficient leading ultimately to loss of utrophin from the membrane and loss of efficacy.…”

Section: Replacement Of Dystrophin With a Surrogate Proteinmentioning

confidence: 99%

From diagnosis to therapy in Duchenne muscular dystrophy

Babbs

Chatzopoulou

Edwards

et al. 2020

Biochemical Society Transactions

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Genetic approaches for the diagnosis and treatment of inherited muscle diseases have advanced rapidly in recent years. Many of the advances have occurred in the treatment of Duchenne muscular dystrophy (DMD), a muscle wasting disease where affected boys are typically wheelchair bound by age 12 years and generally die in their twenties from respiratory failure or cardiomyopathy. Dystrophin is a 421 kD protein which links F-actin to the extracellular matrix via the dystrophin-associated protein complex (DAPC) at the muscle membrane. In the absence of dystrophin, the DAPC is lost, making the muscle membrane more susceptible to contraction-induced injury. The identification of the gene causing DMD in 1986 resulted in improved diagnosis of the disease and the identification of hotspots for mutation. There is currently no effective treatment. However, there are several promising genetic therapeutic approaches at the preclinical stage or in clinical trials including read-through of stop codons, exon skipping, delivery of dystrophin minigenes and the modulation of expression of the dystrophin related protein, utrophin. In spite of significant progress, the problem of targeting all muscles, including diaphragm and heart at sufficiently high levels, remains a challenge. Any therapy also needs to consider the immune response and some treatments are mutation specific and therefore limited to a subgroup of patients. This short review provides a summary of the current status of DMD therapy with a particular focus on those genetic strategies that have been taken to the clinic.

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Isolation, Structural Identification, Synthesis, and Pharmacological Profiling of 1,2-trans-Dihydro-1,2-diol Metabolites of the Utrophin Modulator Ezutromid

Cited by 10 publications

References 38 publications

High-throughput screening identifies modulators of sarcospan that stabilize muscle cells and exhibit activity in the mouse model of Duchenne muscular dystrophy

High-throughput screening identifies modulators of sarcospan that stabilize muscle cells and exhibit activity in the mouse model of Duchenne muscular dystrophy

Utrophin modulator drugs as potential therapies for Duchenne and Becker muscular dystrophies

From diagnosis to therapy in Duchenne muscular dystrophy

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