{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T02:35:00Z","timestamp":1760150100708,"version":"build-2065373602"},"reference-count":40,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2023,10,13]],"date-time":"2023-10-13T00:00:00Z","timestamp":1697155200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Australian Institute for Musculoskeletal Science"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Adenylosuccinic acid (ASA) is a small molecule dicarboxylate that could be a strong clinical development candidate for inherited myopathies involving dysregulated purine nucleotide metabolism. Currently, there are no published pharmacokinetic\/dynamic or toxicology data available, although 10-year clinical trial data on Duchenne muscular dystrophy patients suggests it is a chronically safe drug. In this study, we tested the toxicity of ASA to cultured myoblasts in vitro and its acute systemic toxicity in mice. ASA is a non-toxic small molecule with an LD50 &gt; 5000 mg\/kg. Some background necrotic foci in the liver, kidney and gastrointestinal tract were shown that are likely incidental but warrant follow-up sub-\/chronic oral exposure studies.<\/jats:p>","DOI":"10.3390\/ph16101458","type":"journal-article","created":{"date-parts":[[2023,10,13]],"date-time":"2023-10-13T10:04:39Z","timestamp":1697191479000},"page":"1458","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Adenylosuccinic Acid Is a Non-Toxic Small Molecule In Vitro and In Vivo"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4567-4319","authenticated-orcid":false,"given":"Cara A.","family":"Timpani","sequence":"first","affiliation":[{"name":"Institute for Health and Sport (IHeS), Victoria University, Melbourne, VIC 8001, Australia"},{"name":"Inherited and Acquired Myopathy Program, Australian Institute for Musculoskeletal Science (AIMSS), St Albans, VIC 3021, Australia"},{"name":"Department of Medicine\u2014Western Health, Melbourne Medical School, The University of Melbourne, St Albans, VIC 3021, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lorna","family":"Rasmussen","sequence":"additional","affiliation":[{"name":"Cerberus Sciences, Scoresby, VIC 3179, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4854-0036","authenticated-orcid":false,"given":"Emma","family":"Rybalka","sequence":"additional","affiliation":[{"name":"Institute for Health and Sport (IHeS), Victoria University, Melbourne, VIC 8001, Australia"},{"name":"Inherited and Acquired Myopathy Program, Australian Institute for Musculoskeletal Science (AIMSS), St Albans, VIC 3021, Australia"},{"name":"Department of Medicine\u2014Western Health, Melbourne Medical School, The University of Melbourne, St Albans, VIC 3021, Australia"},{"name":"Division of Neuropaediatrics and Developmental Medicine, University Children\u2019s Hospital of Basel (UKBB), 4031 Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/0306-9877(92)90087-S","article-title":"The dystrophin connection\u2014ATP?","volume":"38","author":"Bonsett","year":"1992","journal-title":"Med. Hypotheses"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1021","DOI":"10.1016\/j.mehy.2015.08.015","article-title":"Revisiting the dystrophin-ATP connection: How half a century of research still implicates mitochondrial dysfunction in Duchenne Muscular Dystrophy aetiology","volume":"85","author":"Timpani","year":"2015","journal-title":"Med. Hypotheses"},{"key":"ref_3","first-page":"157","article-title":"The role of mitochondria in Duchenne muscular dystrophy","volume":"72","author":"Budzinska","year":"2021","journal-title":"J. Physiol. Pharmacol."},{"key":"ref_4","first-page":"446","article-title":"Duchenne\u2019s muscular dystrophy: A tissue culture perspective","volume":"77","author":"Bonsett","year":"1984","journal-title":"Indiana Med. J. Indiana State Med. Assoc."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Chandel, N.S. (2021). Nucleotide metabolism. Cold Spring Harb. Perspect. Biol., 13.","DOI":"10.1101\/cshperspect.a040592"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1002\/bmb.2005.494033032469","article-title":"Facilitating understanding of the purine nucleotide cycle and the one-carbon pool: Part I: The purine nucleotide cycle","volume":"33","author":"Arinze","year":"2005","journal-title":"Biochem. Mol. Biol. Educ."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1422","DOI":"10.1172\/JCI111553","article-title":"Disruption of the purine nucleotide cycle by inhibition of adenylosuccinate lyase produces skeletal muscle dysfunction","volume":"74","author":"Swain","year":"1984","journal-title":"J. Clin. Investig."},{"key":"ref_8","first-page":"2308","article-title":"The purine nucleotide cycle in skeletal muscle","volume":"37","author":"Lowenstein","year":"1978","journal-title":"Fed. Proc."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Rybalka, E., Kourkais, S., Bonsett, C.A., Moghadaszadeh, B., Beggs, A.H., and Timpani, C.A. (2023). Adenylosuccinic Acid: An Orphan Drug with Untapped Potential. Pharmaceuticals, 16.","DOI":"10.3390\/ph16060822"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/j.celrep.2015.08.072","article-title":"Adenylosuccinate is an insulin secretagogue derived from glucose-induced purine metabolism","volume":"13","author":"Gooding","year":"2015","journal-title":"Cell Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/0306-9877(94)90112-0","article-title":"\u2018Oil globules\u2019 in Duchenne muscular dystrophy\u2014History, demonstration, metabolic significance","volume":"43","author":"Bonsett","year":"1994","journal-title":"Med. Hypotheses"},{"key":"ref_12","first-page":"362","article-title":"From idea to market: The drug approval process","volume":"14","author":"Lipsky","year":"2001","journal-title":"J. Am. Board. Fam. Pract."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1016\/j.jacbts.2016.06.003","article-title":"Drugs and devices: Comparison of European and US approval processes","volume":"1","year":"2016","journal-title":"JACC Basic. Transl. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3051","DOI":"10.1016\/j.jaip.2022.09.005","article-title":"Overview of FDA drug approval and labeling","volume":"10","author":"Clarridge","year":"2022","journal-title":"J. Allergy Clin. Immunol. Pract."},{"key":"ref_15","unstructured":"OECD (2022). Test No. 425: Acute Oral Toxicity: Up-and-Down Procedure, OECD."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Mazzaccara, C., Labruna, G., Cito, G., Scarfo, M., De Felice, M., Pastore, L., and Sacchetti, L. (2008). Age-Related Reference Intervals of the Main Biochemical and Hematological Parameters in C57BL\/6J, 129SV\/EV and C3H\/HeJ Mouse Strains. PLoS ONE, 3.","DOI":"10.1371\/journal.pone.0003772"},{"key":"ref_17","first-page":"387","article-title":"Reference values for serum proteins of common laboratory rodent strains","volume":"48","author":"Zaias","year":"2009","journal-title":"J. Am. Assoc. Lab. Anim. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1002\/ame2.12139","article-title":"Clinical hematological and biochemical parameters in Swiss, BALB\/c, C57BL\/6 and B6D2F1 Mus musculus","volume":"3","author":"Bax","year":"2020","journal-title":"Anim. Models Exp. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1007\/PL00000229","article-title":"Differences in normal values for murine white blood cell counts and other hematological parameters based on sampling site","volume":"50","author":"Nemzek","year":"2001","journal-title":"Inflamm. Res."},{"key":"ref_20","unstructured":"McInnes, E.F. (2012). Background Lesions in Laboratory Animals: A Color Atlas, Saunders\/Elsevier."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1080\/03007995.2020.1865699","article-title":"Adenylosuccinic acid: A novel inducer of the cytoprotectant Nrf2 with efficacy in Duchenne muscular dystrophy","volume":"37","author":"Rybalka","year":"2021","journal-title":"Curr. Med. Res. Opin."},{"key":"ref_22","first-page":"2752","article-title":"The role of Nrf2\/PIWIL2\/purine metabolism axis in controlling radiation-induced lung fibrosis","volume":"10","author":"Zou","year":"2020","journal-title":"Am. J. Cancer Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"E827","DOI":"10.1152\/ajpendo.00378.2020","article-title":"New insights into purine metabolism in metabolic diseases: Role of xanthine oxidoreductase activity","volume":"319","author":"Furuhashi","year":"2020","journal-title":"Am. J. Physiol. Endocrinol. Metab."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"278","DOI":"10.1016\/j.cell.2019.12.017","article-title":"FAMIN Is a Multifunctional Purine Enzyme Enabling the Purine Nucleotide Cycle","volume":"180","author":"Cader","year":"2020","journal-title":"Cell"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Kourakis, S., Timpani, C.A., de Haan, J.B., Gueven, N., Fischer, D., and Rybalka, E. (2020). Dimethyl Fumarate and Its Esters: A Drug with Broad Clinical Utility?. Pharmaceuticals, 13.","DOI":"10.20944\/preprints202009.0684.v1"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"630","DOI":"10.1111\/j.0007-0963.2004.05903.x","article-title":"Fumaric acid esters, their place in the treatment of psoriasis","volume":"150","author":"Ormerod","year":"2004","journal-title":"Br. J. Dermatol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1111\/j.1365-2133.2005.06728.x","article-title":"Fumaric acid esters for severe psoriasis: A retrospective review of 58 cases","volume":"153","author":"Harries","year":"2005","journal-title":"Br. J. Dermatol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1007\/s11845-016-1470-2","article-title":"Fumaric acid esters for psoriasis: A systematic review","volume":"186","author":"Smith","year":"2017","journal-title":"Ir. J. Med. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3154","DOI":"10.1007\/s12325-019-01085-3","article-title":"Improving the Gastrointestinal Tolerability of Fumaric Acid Esters: Early Findings on Gastrointestinal Events with Diroximel Fumarate in Patients with Relapsing-Remitting Multiple Sclerosis from the Phase 3, Open-Label EVOLVE-MS-1 Study","volume":"36","author":"Palte","year":"2019","journal-title":"Adv. Ther."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"9","DOI":"10.7224\/1537-2073.2014-101","article-title":"Gastrointestinal tolerability of delayed-release dimethyl fumarate in a multicenter, open-label study of patients with relapsing forms of multiple sclerosis (MANAGE)","volume":"18","author":"Fox","year":"2016","journal-title":"Int. J. MS Care"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"107710","DOI":"10.1016\/j.pharmthera.2020.107710","article-title":"Dimethyl fumarate induced lymphopenia in multiple sclerosis: A review of the literature","volume":"219","author":"Russo","year":"2021","journal-title":"Pharmacol. Ther."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.1039\/C6MD00005C","article-title":"SLC Transporters: Structure, Function, Drug Discovery","volume":"7","author":"Colas","year":"2016","journal-title":"Medchemcomm"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"618411","DOI":"10.3389\/fphar.2021.618411","article-title":"Advances in Oral Drug Delivery","volume":"12","author":"Alqahtani","year":"2021","journal-title":"Front. Pharmacol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"481","DOI":"10.7861\/clinmedicine.16-5-481","article-title":"Adverse drug reactions","volume":"16","author":"Coleman","year":"2016","journal-title":"Clin. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3","DOI":"10.2133\/dmpk.DMPK-10-RV-062","article-title":"Mechanisms of drug toxicity and relevance to pharmaceutical development","volume":"26","author":"Guengerich","year":"2011","journal-title":"Drug Metab. Pharmacokinet."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1125","DOI":"10.1038\/s41598-020-57610-w","article-title":"Adenylosuccinic acid therapy ameliorates murine Duchenne Muscular Dystrophy","volume":"10","author":"Timpani","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_37","first-page":"2055217315596994","article-title":"Dimethyl fumarate-associated lymphopenia: Risk factors and clinical significance","volume":"1","author":"Longbrake","year":"2015","journal-title":"Mult. Scler. J.\u2013Exp. Transl. Clin."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Timpani, C.A., and Rybalka, E. (2021). Calming the (Cytokine) Storm: Dimethyl Fumarate as a Therapeutic Candidate for COVID-19. Pharmaceuticals, 14.","DOI":"10.3390\/ph14010015"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Timpani, C.A., Mamchaoui, K., Butler-Browne, G., and Rybalka, E. (2020). Nitric Oxide (NO) and Duchenne Muscular Dystrophy: NO Way to Go?. Antioxidants, 9.","DOI":"10.3390\/antiox9121268"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.mito.2017.10.010","article-title":"Chemotherapeutic agents induce mitochondrial superoxide production and toxicity but do not alter respiration in skeletal muscle in vitro","volume":"42","author":"Rybalka","year":"2018","journal-title":"Mitochondrion"}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/16\/10\/1458\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:06:32Z","timestamp":1760130392000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/16\/10\/1458"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10,13]]},"references-count":40,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["ph16101458"],"URL":"https:\/\/doi.org\/10.3390\/ph16101458","relation":{},"ISSN":["1424-8247"],"issn-type":[{"type":"electronic","value":"1424-8247"}],"subject":[],"published":{"date-parts":[[2023,10,13]]}}}