{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T04:15:23Z","timestamp":1787804123049,"version":"build-2784847793"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T00:00:00Z","timestamp":1753401600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"\n Background<\/jats:title>\n Zagociguat (zag) is a CNS-penetrant, soluble guanylate cyclase (sGC) stimulator that has been evaluated in phase 2a, with phase 2b ongoing, clinical studies of primary mitochondrial disease (PMD) subjects with mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS). To explore its utility in a broader array of PMDs and secondary mitochondrial disorders, we performed prfeclinical modeling of zag across larval and adult zebrafish models with biochemical deficiencies in diverse respiratory chain (RC) complexes or dihydrolipoamide dehydrogenase (Dldh).<\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n Zag was evaluated for tissue uptake, gross toxicity, protection from RC toxin-induced brain death, neuromuscular dysfunction, heartbeat loss, and biochemical dysfunction in transgenic or toxin-exposed zebrafish with mitochondrial enzyme deficiencies in complex I (\n ndufs2<\/jats:italic>\n \n \u2212\/\u2212<\/jats:italic>\n <\/jats:sup>\n or rotenone-exposed wild type (WT)), complex IV (\n surf1<\/jats:italic>\n \n \u2212\/\u2212<\/jats:italic>\n <\/jats:sup>\n or azide-exposed WT), multiple RC complexes (\n fbxl4<\/jats:italic>\n \n \u2212\/\u2212<\/jats:italic>\n <\/jats:sup>\n ), or pyruvate dehydrogenase complex (\n dldh<\/jats:italic>\n \n \u2212\/\u2212<\/jats:italic>\n <\/jats:sup>\n ). Zag effects were also studied on the whole-body oxygen consumption capacity (MO\n 2<\/jats:sub>\n ) and swimming activity of WT and complex IV disease adult zebrafish.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Similar zag levels were observed in adult brains and tail muscle. No morphological or functional toxic effects of zag were observed on larvae viability. Zag provided neuromuscular protection in complex I deficient genetic and pharmacologic inhibitor models. In complex IV deficient models, prevention from brain death occurred at 100\u00a0nM zag in high-dose azide-exposed WT larvae; however, no rescue of swimming or neuromuscular phenotypes in low-dose azide-exposed\n surf1<\/jats:italic>\n \n \u2212\/\u2212<\/jats:italic>\n <\/jats:sup>\n larvae was observed. A total of 100\u00a0nM zag rescued MO\n 2<\/jats:sub>\n and maximum swimming speed in adult\n surf1<\/jats:italic>\n \n \u2212\/\u2212<\/jats:italic>\n <\/jats:sup>\n zebrafish. Larval swimming activity was also preserved with 10\u00a0nM zag treatment in azide-stressed\n fbxl4<\/jats:italic>\n \n \u2212\/\u2212<\/jats:italic>\n <\/jats:sup>\n larvae but not at 10\u00a0nM, 100\u00a0nM, or 1\u00a0\u00b5M zag in\n dldh<\/jats:italic>\n \n \u2212\/\u2212<\/jats:italic>\n <\/jats:sup>\n larvae. Zag (10\u00a0nM) enhanced complex I enzyme activity that is suggestive of mitochondrial biogenesis and key aspects of mitochondrial physiology in azide-exposed\n surf1<\/jats:italic>\n \n \u2212\/\u2212<\/jats:italic>\n <\/jats:sup>\n and\n fbxl4<\/jats:italic>\n \u2212\/\u2212<\/jats:sup>\n larvae.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n Preclinical evaluation of zag demonstrated its safety, significant protection of neuromuscular dysfunction and\/or acute RC stressor-induced decompensation, and improved mitochondrial physiology across multiple different genetic and\/or pharmacologic models of RC-deficient PMD. Thus, zag may yield therapeutic potential for an array of diseases with mitochondrial dysfunction beyond MELAS, potentially including Leigh syndrome spectrum disorder and primary mitochondrial myopathies.<\/jats:p>\n <\/jats:sec>","DOI":"10.3389\/fphar.2025.1588426","type":"journal-article","created":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T09:45:37Z","timestamp":1753436737000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Zagociguat prevented stressor-induced neuromuscular dysfunction, improved mitochondrial physiology, and increased exercise capacity in diverse mitochondrial respiratory chain disease zebrafish models"],"prefix":"10.3389","volume":"16","author":[{"given":"Leonard","family":"Burg","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heeyong","family":"Yoon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Peng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Germano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily","family":"Reesey Gretzmacher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vernon E.","family":"Anderson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eiko","family":"Nakamaru-Ogiso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marni J.","family":"Falk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,7,25]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"236","DOI":"10.1002\/path.4809","article-title":"The genetics and pathology of mitochondrial disease","volume":"241","author":"Alston","year":"2017","journal-title":"J. Pathol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1097\/MOP.0000000000000954","article-title":"Mitochondrial medicine therapies: rationale, evidence, and dosing guidelines","volume":"32","author":"Barcelos","year":"2020","journal-title":"Curr. Opin. Pediatr."},{"key":"B35","doi-asserted-by":"publisher","first-page":"1407","DOI":"10.1007\/s40262-020-00895-x","article-title":"Metabolism and pharmacokinetic drug-drug interaction profile of vericiguat, a soluble guanylate cyclase stimulator: results from preclinical and phase I healthy volunteer studies","volume":"59","author":"Boettcher","year":"2020","journal-title":"Clin. Pharmacokinet."},{"key":"B3","doi-asserted-by":"publisher","first-page":"801","DOI":"10.3390\/ijms19030801","article-title":"cGMP signaling in the cardiovascular system-the role of compartmentation and its live cell imaging","volume":"19","author":"Bork","year":"2018","journal-title":"Int. J. Mol. Sci."},{"key":"B4","doi-asserted-by":"publisher","first-page":"248","DOI":"10.1006\/abio.1976.9999","article-title":"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem."},{"key":"B5","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1016\/j.neuint.2017.07.008","article-title":"Pharmacologic modeling of primary mitochondrial respiratory chain dysfunction in zebrafish","volume":"117","author":"Byrnes","year":"2018","journal-title":"Neurochem. Int."},{"key":"B6","doi-asserted-by":"publisher","first-page":"656561","DOI":"10.3389\/fphar.2021.656561","article-title":"The CNS-penetrant soluble guanylate cyclase stimulator CY6463 reveals its therapeutic potential in neurodegenerative diseases","volume":"12","author":"Correia","year":"2021","journal-title":"Front. Pharmacol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"247","DOI":"10.1016\/j.ymgme.2012.06.018","article-title":"Citrulline and arginine utility in treating nitric oxide deficiency in mitochondrial disorders","volume":"107","author":"El-Hattab","year":"2012","journal-title":"Mol. Genet. Metab."},{"key":"B8","doi-asserted-by":"publisher","first-page":"407","DOI":"10.1016\/j.ymgme.2016.01.010","article-title":"Impaired nitric oxide production in children with MELAS syndrome and the effect of arginine and citrulline supplementation","volume":"117","author":"El-Hattab","year":"2016","journal-title":"Mol. Genet. Metab."},{"key":"B9","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1002\/crt2.41","article-title":"Development of a mitochondrial myopathy-composite assessment tool","volume":"6","author":"Flickinger","year":"2021","journal-title":"JCSM Clin. Rep."},{"key":"B10","doi-asserted-by":"publisher","first-page":"1546","DOI":"10.1074\/jbc.M114.579565","article-title":"Increasing NAD synthesis in muscle via nicotinamide phosphoribosyltransferase is not sufficient to promote oxidative metabolism","volume":"290","author":"Frederick","year":"2015","journal-title":"J. Biol. Chem."},{"key":"B36","doi-asserted-by":"publisher","first-page":"647","DOI":"10.1007\/s40262-017-0604-7","article-title":"Clinical pharmacokinetic and pharmacodynamic profile of riociguat","volume":"57","author":"Frey","year":"2018","journal-title":"Clin. Pharmacokinet."},{"key":"B11","doi-asserted-by":"publisher","first-page":"482","DOI":"10.1016\/j.ajhg.2013.07.016","article-title":"Mutations in FBXL4, encoding a mitochondrial protein, cause early-onset mitochondrial encephalomyopathy","volume":"93","author":"Gai","year":"2013","journal-title":"Am. J. Hum. Genet."},{"key":"B12","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1016\/j.ymgme.2018.01.010","article-title":"8-year retrospective analysis of intravenous arginine therapy for acute metabolic strokes in pediatric mitochondrial disease","volume":"123","author":"Ganetzky","year":"","journal-title":"Mol. Genet. Metab."},{"key":"B13","doi-asserted-by":"publisher","first-page":"74","DOI":"10.1016\/j.ymgmr.2018.03.001","article-title":"Microvascular endothelial dysfunction in mitochondrial stroke-like episodes supports use of intravenous l-arginine","volume":"15","author":"Ganetzky","year":"","journal-title":"Mol. Genet. Metab. Rep."},{"key":"B14","doi-asserted-by":"publisher","first-page":"448a","DOI":"10.1016\/j.bpj.2024.11.2387","article-title":"BPS2025 - mechanistic investigation of sAC-dependent PKA activation in mitochondrial dysfunction","volume":"124","author":"Gest","year":"2025","journal-title":"Biophysical J."},{"key":"B15","doi-asserted-by":"publisher","first-page":"536","DOI":"10.1093\/hmg\/ddab059","article-title":"Combinatorial glucose, nicotinic acid and N-acetylcysteine therapy has synergistic effect in preclinical C. elegans and zebrafish models of mitochondrial complex I disease","volume":"30","author":"Guha","year":"2021","journal-title":"Hum. Mol. Genet."},{"key":"B16","doi-asserted-by":"publisher","first-page":"5323","DOI":"10.1021\/acs.jmedchem.0c02154","article-title":"Discovery of the soluble guanylate cyclase activator runcaciguat (BAY 1101042)","volume":"64","author":"Hahn","year":"2021","journal-title":"J. Med. Chem."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1988","DOI":"10.1093\/hmg\/ddad031","article-title":"N-Acetylcysteine and cysteamine bitartrate prevent azide-induced neuromuscular decompensation by restoring glutathione balance in two novel surf1-\/- zebrafish deletion models of leigh syndrome","volume":"32","author":"Haroon","year":"2023","journal-title":"Hum. Mol. Genet."},{"key":"B18","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1016\/j.brainres.2004.03.004","article-title":"Effect of protein kinases on lactate dehydrogenase activity in cortical neurons during hypoxia","volume":"1009","author":"Hong","year":"2004","journal-title":"Brain Res."},{"key":"B19","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1097\/MCO.0000000000000610","article-title":"Arginine therapy in mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes","volume":"23","author":"Ikawa","year":"2020","journal-title":"Curr. Opin. Clin. Nutr. Metab. Care"},{"key":"B20","doi-asserted-by":"publisher","first-page":"230","DOI":"10.1016\/j.yjmcc.2008.05.012","article-title":"Cyclic GMP signaling in cardiomyocytes modulates fatty acid trafficking and prevents triglyceride accumulation","volume":"45","author":"Khairallah","year":"2008","journal-title":"J. Mol. Cell Cardiol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"e178973","DOI":"10.1172\/jci.insight.178973","article-title":"dldhcri3 zebrafish exhibit altered mitochondrial ultrastructure, morphology, and dysfunction partially rescued by probucol or thiamine","volume":"9","author":"Lavorato","year":"2024","journal-title":"JCI Insight"},{"key":"B22","doi-asserted-by":"publisher","first-page":"e156346","DOI":"10.1172\/jci.insight.156346","article-title":"Dichloroacetate improves mitochondrial function, physiology, and morphology in FBXL4 disease models","volume":"7","author":"Lavorato","year":"2022","journal-title":"JCI Insight"},{"key":"B23","doi-asserted-by":"publisher","first-page":"721","DOI":"10.1007\/s40265-021-01496-z","article-title":"Vericiguat: first approval","volume":"81","author":"Markham","year":"2021","journal-title":"Drugs"},{"key":"B24","doi-asserted-by":"publisher","first-page":"714","DOI":"10.1097\/MOP.0000000000000686","article-title":"Mitochondrial disease genetics update: recent insights into the molecular diagnosis and expanding phenotype of primary mitochondrial disease","volume":"30","author":"McCormick","year":"2018","journal-title":"Curr. Opin. Pediatr."},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1101\/2025.07.16.664929","article-title":"ndufs2-\/- zebrafish have impaired survival, neuromuscular activity, morphology, and one-carbon metabolism treatable with folic acid","author":"Mitchell","year":"2025","journal-title":"BioRxiv."},{"key":"B25","doi-asserted-by":"publisher","first-page":"2235","DOI":"10.1056\/NEJMoa2504747","article-title":"Patient-specific in vivo gene editing to treat a rare genetic disease","volume":"392","author":"Musunuru","year":"2025","journal-title":"N. Engl. J. Med."},{"key":"B26","doi-asserted-by":"publisher","first-page":"719","DOI":"10.1016\/j.bbabio.2008.04.005","article-title":"Mammalian complex I: a regulable and vulnerable pacemaker in mitochondrial respiratory function","volume":"1777","author":"Papa","year":"2008","journal-title":"Biochim. Biophys. Acta"},{"key":"B28","doi-asserted-by":"publisher","first-page":"2258","DOI":"10.1021\/bi201573k","article-title":"Proteolytic regulation of the mitochondrial cAMP-dependent protein kinase","volume":"51","author":"Shell","year":"2012","journal-title":"Biochemistry"},{"key":"B29","doi-asserted-by":"publisher","first-page":"9655","DOI":"10.3390\/ijms23179655","article-title":"cAMP\/PKA signaling modulates mitochondrial supercomplex organization","volume":"23","author":"Signorile","year":"2022","journal-title":"Int. J. Mol. Sci."},{"key":"B30","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1042\/BST20110655","article-title":"cGMP-cAMP interplay in cardiac myocytes: a local affair with far-reaching consequences for heart function","volume":"40","author":"Stangherlin","year":"2012","journal-title":"Biochem. Soc. Trans."},{"key":"B31","doi-asserted-by":"publisher","first-page":"3606","DOI":"10.1111\/bcp.15861","article-title":"Randomized placebo-controlled crossover study to assess tolerability and pharmacodynamics of zagociguat, a soluble guanylyl cyclase stimulator, in healthy elderly","volume":"89","author":"van Kraaij","year":"","journal-title":"Br. J. Clin. Pharmacol."},{"key":"B32","doi-asserted-by":"publisher","first-page":"1381","DOI":"10.1111\/cts.13537","article-title":"First-in-human trial to assess safety, tolerability, pharmacokinetics, and pharmacodynamics of zagociguat (CY6463), a CNS-Penetrant soluble guanylyl cyclase stimulator","volume":"16","author":"van Kraaij","year":"","journal-title":"Clin. Transl. Sci."},{"key":"B33","volume-title":"The zebrafish book; A guide for the laboratory use of zebrafish (","author":"Westerfield","year":"2007"},{"key":"B34","doi-asserted-by":"publisher","first-page":"e0197513","DOI":"10.1371\/journal.pone.0197513","article-title":"Mitochondrial disease patient motivations and barriers to participate in clinical trials","volume":"13","author":"Zolkipli-Cunningham","year":"2018","journal-title":"PLoS One"}],"container-title":["Frontiers in Pharmacology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2025.1588426\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T09:45:38Z","timestamp":1753436738000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2025.1588426\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,25]]},"references-count":36,"alternative-id":["10.3389\/fphar.2025.1588426"],"URL":"https:\/\/doi.org\/10.3389\/fphar.2025.1588426","relation":{},"ISSN":["1663-9812"],"issn-type":[{"value":"1663-9812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,25]]},"article-number":"1588426"}}