{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T03:26:18Z","timestamp":1761017178125},"reference-count":29,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,3,22]],"date-time":"2017-03-22T00:00:00Z","timestamp":1490140800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,3,22]],"date-time":"2017-03-22T00:00:00Z","timestamp":1490140800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Severe bradycardia\/bradyarrhythmia following coadministration of the HCV-NS5B prodrug sofosbuvir with amiodarone was recently reported. Our previous preclinical <jats:italic>in vivo<\/jats:italic> experiments demonstrated that only certain HCV-NS5B prodrugs elicit bradycardia when combined with amiodarone. In this study, we evaluate the impact of HCV-NS5B prodrug phosphoramidate diastereochemistry (D-\/L-alanine, R-\/S-phosphoryl) <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic>. Co-applied with amiodarone, L-ala,<jats:italic>S<\/jats:italic><jats:sub>P<\/jats:sub> prodrugs increased beating rate and decreased beat amplitude in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), but D-ala,<jats:italic>R<\/jats:italic><jats:sub>P<\/jats:sub> produgs, including MK-3682, did not. Stereochemical selectivity on emerging bradycardia was confirmed <jats:italic>in vivo<\/jats:italic>. Diastereomer pairs entered cells equally well, and there was no difference in intracellular accumulation of L-ala,<jats:italic>S<\/jats:italic><jats:sub>P<\/jats:sub> metabolites\u2009\u00b1\u2009amiodarone, but no D-ala,<jats:italic>R<\/jats:italic><jats:sub>P<\/jats:sub> metabolites were detected. Cathepsin A (CatA) inhibitors attenuated L-ala,<jats:italic>S<\/jats:italic><jats:sub>P<\/jats:sub> prodrug metabolite formation, yet exacerbated L-ala,<jats:italic>S<\/jats:italic><jats:sub>P<\/jats:sub>\u2009+\u2009amiodarone effects, implicating the prodrugs in these effects. Experiments indicate that pharmacological effects and metabolic conversion to UTP analog are L-ala,<jats:italic>S<\/jats:italic><jats:sub>P<\/jats:sub> prodrug-dependent in cardiomyocytes.<\/jats:p>","DOI":"10.1038\/srep44820","type":"journal-article","created":{"date-parts":[[2017,3,22]],"date-time":"2017-03-22T10:37:12Z","timestamp":1490179032000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Cardiac drug-drug interaction between HCV-NS5B pronucleotide inhibitors and amiodarone is determined by their specific diastereochemistry"],"prefix":"10.1038","volume":"7","author":[{"given":"Armando","family":"Lagrutta","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher P.","family":"Regan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haoyu","family":"Zeng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John P.","family":"Imredy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kenneth","family":"Koeplinger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pierre","family":"Morissette","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Liping","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gordon","family":"Wollenberg","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher","family":"Brynczka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jos\u00e9","family":"Lebr\u00f3n","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph","family":"DeGeorge","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frederick","family":"Sannajust","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2017,3,22]]},"reference":[{"key":"BFsrep44820_CR1","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1002\/hep.27488","volume":"62","author":"T Ahmad","year":"2015","unstructured":"Ahmad, T. et al. Cardiac dysfunction associated with a nucleotide polymerase inhibitor for treatment of hepatitis C. Hepatology 62, 409\u2013416 (2015).","journal-title":"Hepatology"},{"key":"BFsrep44820_CR2","first-page":"221","volume":"138","author":"J Kwagh","year":"2014","unstructured":"Kwagh, J. et al. BMS-986094: Potential Cytotoxicity in Differentiated Human Cardiomyocytes. The Toxicologist: Supplement to Toxicological Sciences. Abstract no. 856 138, 221 (2014).","journal-title":"The Toxicologist: Supplement to Toxicological Sciences."},{"key":"BFsrep44820_CR3","doi-asserted-by":"crossref","unstructured":"Baumgart, B. R. et al. Effects of BMS-986094, a Guanosine Nucleotide Analogue, on Mitochondrial DNA Synthesis and Function. Toxicol. Sci. (2016).","DOI":"10.1093\/toxsci\/kfw135"},{"key":"BFsrep44820_CR4","doi-asserted-by":"publisher","first-page":"1886","DOI":"10.1056\/NEJMc1505967","volume":"373","author":"H Fontaine","year":"2015","unstructured":"Fontaine, H. et al. Bradyarrhythmias Associated with Sofosbuvir Treatment. N. Engl. J. Med. 373, 1886\u20131888 (2015).","journal-title":"N. Engl. J. Med."},{"key":"BFsrep44820_CR5","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1016\/j.taap.2016.08.006","volume":"308","author":"A Lagrutta","year":"2016","unstructured":"Lagrutta, A. et al. Interaction between amiodarone and hepatitis-C virus nucleotide inhibitors in human induced pluripotent stem cell-derived cardiomyocytes and HEK-293 Cav1.2 over-expressing cells. Toxicol. Appl. Pharmacol. 308, 66\u201376 (2016).","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"BFsrep44820_CR6","doi-asserted-by":"publisher","first-page":"174","DOI":"10.1093\/toxsci\/kfw153","volume":"154","author":"D Millard","year":"2016","unstructured":"Millard, D. et al. Identification of drug-drug interactions in vitro: a case study evaluating the effects of sofosbuvir and amiodarone on hiPSC-derived cardiomyocytes. Toxicol. Sci. 154, 174\u2013182 (2016).","journal-title":"Toxicol. Sci."},{"key":"BFsrep44820_CR7","doi-asserted-by":"publisher","first-page":"1430","DOI":"10.1002\/hep.28752","volume":"64","author":"CP Regan","year":"2016","unstructured":"Regan, C. P. et al. Preclinical Assessment of the Clinical Cardiac Drug-Drug Interaction Associated with the Combination of HCV-NI Antivirals and Amiodarone. Hepatology 64, 1430\u20131441 (2016).","journal-title":"Hepatology"},{"key":"BFsrep44820_CR8","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1016\/j.cbi.2006.07.001","volume":"162","author":"T Satoh","year":"2006","unstructured":"Satoh, T. & Hosokawa, M. Structure, function and regulation of carboxylesterases. Chem. Biol. Interact. 162, 195\u2013211 (2006).","journal-title":"Chem. Biol. Interact."},{"key":"BFsrep44820_CR9","doi-asserted-by":"publisher","first-page":"210","DOI":"10.1002\/phar.1194","volume":"33","author":"SC Laizure","year":"2013","unstructured":"Laizure, S. C., Herring, V., Hu, Z., Witbrodt, K. & Parker, R. B. The role of human carboxylesterases in drug metabolism: have we overlooked their importance? Pharmacotherapy 33, 210\u2013222 (2013).","journal-title":"Pharmacotherapy"},{"key":"BFsrep44820_CR10","doi-asserted-by":"publisher","first-page":"34337","DOI":"10.1074\/jbc.M110.161802","volume":"285","author":"E Murakami","year":"2010","unstructured":"Murakami, E. et al. Mechanism of activation of PSI-7851 and its diastereoisomer PSI-7977. J. Biol. Chem. 285, 34337\u201334347 (2010).","journal-title":"J. Biol. Chem."},{"key":"BFsrep44820_CR11","doi-asserted-by":"publisher","first-page":"1734","DOI":"10.1124\/dmd.106.009381","volume":"34","author":"T Imai","year":"2006","unstructured":"Imai, T., Taketani, M., Shii, M., Hosokawa, M. & Chiba, K. Substrate specificity of carboxylesterase isozymes and their contribution to hydrolase activity in human liver and small intestine. Drug Metab Dispos. 34, 1734\u20131741 (2006).","journal-title":"Drug Metab Dispos"},{"key":"BFsrep44820_CR12","doi-asserted-by":"publisher","first-page":"541","DOI":"10.1124\/dmd.30.5.541","volume":"30","author":"M Xie","year":"2002","unstructured":"Xie, M., Yang, D., Liu, L., Xue, B. & Yan, B. Human and rodent carboxylesterases: immunorelatedness, overlapping substrate specificity, differential sensitivity to serine enzyme inhibitors, and tumor-related expression. Drug Metab Dispos. 30, 541\u2013547 (2002).","journal-title":"Drug Metab Dispos"},{"key":"BFsrep44820_CR13","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1007\/s40262-015-0261-7","volume":"54","author":"BJ Kirby","year":"2015","unstructured":"Kirby, B. J., Symonds, W. T., Kearney, B. P. & Mathias, A. A. Pharmacokinetic, Pharmacodynamic, and Drug-Interaction Profile of the Hepatitis C Virus NS5B Polymerase Inhibitor Sofosbuvir. Clin. Pharmacokinet. 54, 677\u2013690 (2015).","journal-title":"Clin. Pharmacokinet."},{"key":"BFsrep44820_CR14","doi-asserted-by":"publisher","first-page":"7202","DOI":"10.1021\/jm100863x","volume":"53","author":"MJ Sofia","year":"2010","unstructured":"Sofia, M. J. et al. Discovery of a beta-d-2\u2032-deoxy-2\u2032-alpha-fluoro-2\u2032-beta-C-methyluridine nucleotide prodrug (PSI-7977) for the treatment of hepatitis C virus. J. Med. Chem. 53, 7202\u20137218 (2010).","journal-title":"J. Med. Chem."},{"key":"BFsrep44820_CR15","doi-asserted-by":"publisher","first-page":"7636","DOI":"10.1021\/jm300663n","volume":"55","author":"S Ruf","year":"2012","unstructured":"Ruf, S. et al. Novel beta-amino acid derivatives as inhibitors of cathepsin A. J. Med. Chem. 55, 7636\u20137649 (2012).","journal-title":"J. Med. Chem."},{"key":"BFsrep44820_CR16","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1002\/cpdd.201","volume":"5","author":"J Tillner","year":"2016","unstructured":"Tillner, J. et al. Tolerability, safety, and pharmacokinetics of the novel cathepsin A inhibitor SAR164653 in healthy subjects. Clin. Pharmacol. Drug Dev. 5, 57\u201368 (2016).","journal-title":"Clin. Pharmacol. Drug Dev"},{"key":"BFsrep44820_CR17","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1186\/s12967-016-0907-8","volume":"14","author":"A Petrera","year":"2016","unstructured":"Petrera, A. et al. Cathepsin A inhibition attenuates myocardial infarction-induced heart failure on the functional and proteomic levels. J. Transl. Med. 14, 153 (2016).","journal-title":"J. Transl. Med."},{"key":"BFsrep44820_CR18","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1080\/03602530600952164","volume":"39","author":"M Hosokawa","year":"2007","unstructured":"Hosokawa, M. et al. Genomic structure and transcriptional regulation of the rat, mouse, and human carboxylesterase genes. Drug Metab Rev. 39, 1\u201315 (2007).","journal-title":"Drug Metab Rev"},{"key":"BFsrep44820_CR19","doi-asserted-by":"publisher","first-page":"976","DOI":"10.1161\/01.HYP.0000017283.67962.02","volume":"39","author":"HL Jackman","year":"2002","unstructured":"Jackman, H. L. et al. Angiotensin 1\u20139 and 1\u20137 release in human heart: role of cathepsin A. Hypertension 39, 976\u2013981 (2002).","journal-title":"Hypertension"},{"key":"BFsrep44820_CR20","doi-asserted-by":"publisher","first-page":"5543","DOI":"10.1073\/pnas.0935295100","volume":"100","author":"ME Mangoni","year":"2003","unstructured":"Mangoni, M. E. et al. Functional role of L-type Cav1.3 Ca2+ channels in cardiac pacemaker activity. Proc. Natl. Acad. Sci. USA 100, 5543\u20135548 (2003).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"BFsrep44820_CR21","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1113\/jphysiol.2004.074047","volume":"562","author":"C Marionneau","year":"2005","unstructured":"Marionneau, C. et al. Specific pattern of ionic channel gene expression associated with pacemaker activity in the mouse heart. J. Physiol 562, 223\u2013234 (2005).","journal-title":"J. Physiol"},{"key":"BFsrep44820_CR22","doi-asserted-by":"publisher","first-page":"19","DOI":"10.3389\/fphys.2015.00019","volume":"6","author":"P Mesirca","year":"2015","unstructured":"Mesirca, P., Torrente, A. G. & Mangoni, M. E. Functional role of voltage gated Ca(2+) channels in heart automaticity. Front Physiol 6, 19 (2015).","journal-title":"Front Physiol"},{"key":"BFsrep44820_CR23","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1038\/nn.2694","volume":"14","author":"SM Baig","year":"2011","unstructured":"Baig, S. M. et al. Loss of Ca(v)1.3 (CACNA1D) function in a human channelopathy with bradycardia and congenital deafness. Nat. Neurosci. 14, 77\u201384 (2011).","journal-title":"Nat. Neurosci."},{"key":"BFsrep44820_CR24","doi-asserted-by":"publisher","first-page":"1301","DOI":"10.1016\/0896-6273(94)90446-4","volume":"12","author":"JP Imredy","year":"1994","unstructured":"Imredy, J. P. & Yue, D. T. Mechanism of Ca(2+)-sensitive inactivation of L-type Ca2+ channels. Neuron 12, 1301\u20131318 (1994).","journal-title":"Neuron"},{"key":"BFsrep44820_CR25","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1146\/annurev.physiol.70.113006.100455","volume":"70","author":"DM Bers","year":"2008","unstructured":"Bers, D. M. Calcium cycling and signaling in cardiac myocytes. Annu. Rev. Physiol 70, 23\u201349 (2008).","journal-title":"Annu. Rev. Physiol"},{"key":"BFsrep44820_CR26","first-page":"S104","volume":"42","author":"P Bhagunde","year":"2015","unstructured":"Bhagunde, P., Rizk, M. L., Marshall, B., Butterton, J. & Gao, W. Viral dynamics modeling of MK-3682 monotherapy in HCV infected patients. Journal of Pharmacokinetics and Pharmacodynamics 42(1) S104\u2013S105 (2015).","journal-title":"Journal of Pharmacokinetics and Pharmacodynamics"},{"key":"BFsrep44820_CR27","doi-asserted-by":"publisher","first-page":"S759","DOI":"10.1016\/S0168-8278(16)01479-3","volume":"64","author":"EJ Gane","year":"2016","unstructured":"Gane, E. J. et al. High efficacy of an 8-week 3-drug regimen of grazoprevir\/MK-8408\/MK-3682 in HCV genotype 1, 2 and 3-infected patients: SVR24 data from the phase 2 C-crest 1 and 2 studies. Journal of Hepatology 64(2) S759 (2016).","journal-title":"Journal of Hepatology"},{"key":"BFsrep44820_CR28","doi-asserted-by":"crossref","unstructured":"Zhang, X. et al. Multi-parametric assessment of cardiomyocyte excitation-contraction coupling using impedance and field potential recording: A tool for cardiac safety assessment. J. Pharmacol. Toxicol. Methods (2016).","DOI":"10.1016\/j.vascn.2016.06.004"},{"key":"BFsrep44820_CR29","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1016\/j.vascn.2015.05.007","volume":"75","author":"P Morissette","year":"2015","unstructured":"Morissette, P. et al. QT interval correction assessment in the anesthetized guinea pig. J. Pharmacol. Toxicol. Methods 75, 52\u201361 (2015).","journal-title":"J. Pharmacol. Toxicol. Methods"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep44820.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep44820","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep44820.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T23:47:27Z","timestamp":1671839247000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep44820"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,3,22]]},"references-count":29,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,4,12]]}},"alternative-id":["BFsrep44820"],"URL":"https:\/\/doi.org\/10.1038\/srep44820","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,3,22]]},"assertion":[{"value":"11 November 2016","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 February 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 March 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"All authors are employees at MRL, Merck & Co, in the preclinical safety assessment area or in the pharmacokinetics, pharmacodynamics, and drug metabolism area. Our work, including investigative efforts such as this one, supports development of proprietary preclinical candidates. The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"44820"}}