Safety risk assessment of in vitro heart in antitumor drug development

doi:10.12307/2024.560

Abstract

Abstract: BACKGROUND: Tyrosine kinase inhibitors, as well as other types of small-molecule cancer drugs, can cause severe cardiotoxicity.
OBJECTIVE: To perform a heart safety re-evaluation by observing the effects of antitumor drugs on isolated heart electrocardiograph, cardiac action potential and associated ion channels and cytotoxicity.
METHODS: Extracorporeal cardiac perfusion was given to the isolated rabbit heart using Langendorff perfusion: Sunitinib (0.3, 3, 10 μmol/L), Crizotinib (0.3, 1, 3 μmol/L), and Doxorubicin (1, 30 μmol/L) were perfused sequentially for 120 minutes to record electrocardiograph and left ventricular pressure. A blank control group was set for comparison. Manual patch clamp was used to record the effects of Crizotinib, Sunitinib, Doxorubicin on hERG, Cav1.2, Nav1.5 channel currents and action potential in human induced pluripotent stem cell derived cardiomyocytes. Adenosine triphosphate level in human induced pluripotent stem cell derived cardiomyocytes was detected by CellTiter-Glo luminescent cell viability assay.
RESULTS AND CONCLUSION: Isolated rabbit heart using Langendorff perfusion: Compared with the blank ontrol group, Sunitinib and Crizotinib at ≥ 3 μmol/L decreased heart rate (P < 0.01) and prolonged QT/QTc interval (P < 0.01), and reduced left ventricular pressure to different extents. Manual patch clamp recording: Compared with the blank control group, Sunitinib and Crizotinib at 3 μmol/L inhibited the activities of hERG, Nav1.5 and Cav1.2 channels and significantly prolonged the duration of action potential (P < 0.01). According to the analysis of the test article, the difference between the labeled concentration and the measured concentration of the recovered solution was not significant. Cell viability assays: Compared with the blank control group, adenosine triphosphate content in human induced pluripotent stem cell derived cardiomyocytes significantly decreased after treatment with Sunitinib (IC50=4.64 μmol/L), Doxorubicin (IC50=4.21 μmol/L) and Crizotinib (IC50=2.87 μmol/L), indicating that cell viability significantly decreased (P < 0.01). To conclude, this study successfully established an early cardiac safety evaluation method for antitumor drugs, which provides good support and help for the subsequent development of antitumor drugs.

Key words: Sunitinib, Crizotinib, Doxorubicin, Langendorff, patch clamp, cardiac safety evaluation

CLC Number:

Zheng Shuangjia, Zhao Ting, Ren Cuixia, Wang Baoqiang, Chen Lanlan, Lin Moxu, Li Yingji, Zhang Xu. Safety risk assessment of in vitro heart in antitumor drug development[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(27): 4265-4272.

Figures/Tables 17

References

[1] 司丹妮,李彩娥,陈馨嵘,等.酪氨酸激酶抑制剂诱导的心脏毒性发病情况、评估和监测研究进展[J]. 山东医药,2022,62(33):97-102.
[2] SELTZER JH, GINTANT G, AMIRI-KORDESTANI L, et al. Assessing cardiac safety in oncology drug development. Am Heart J. 2019;214:125.
[3] SHYAM SUNDER S, SHARMA UC, POKHAREL S. Adverse effects of tyrosine kinase inhibitors in cancer therapy: pathophysiology, mechanisms and clinical management. Signal Transduct Target Ther. 2023;8(1):262
[4] WANG H, WANG Y, LI J, et al. Three tyrosine kinase inhibitors cause cardiotoxicity by inducing endoplasmic reticulum stress and inflammation in cardiomyocytes. BMC Med. 2023;21(1):147
[5] KADDOURA R, DABDOOB WA, AHMED K, et al. A practical guide to managing cardiopulmonary toxicities of tyrosine kinase inhibitors in chronic myeloid leukemia. Front Med (Lausanne). 2023;10:1163137.
[6] YAMAMICHI Y, KUSUOKA H, MORISHITA K, et al. Metabolism of iodine-123-BMIPP in perfused rat hearts. J Nucl Med. 1995;36(6):1043-1050.
[7] RUMSEY WL, PATEL B, LINDER KE. Effect of graded hypoxia on retention of technetium-99m-nitroheterocycle in perfused rat heart. J Nucl Med. 1995;36(4): 632-636.
[8] KUBURAS R, GHARANEI M, HAUSSMANN I, et al. Metformin protects against sunitinib-induced cardiotoxicity: investigating the role of AMPK. J Cardiovasc Pharmacol. 2022;79(6):799-807.
[9] OBEJERO-PAZ CA, BRUENING-WRIGHT A, KRAMER J, et al. Quantitative profiling of the effects of vanoxerine on human cardiac ion channels and its application to cardiac risk. Sci Rep. 2015;5:17623.
[10] JONSSON MK, DUKER G, TROPP C, et al. Quantified proarrhythmic potential of selected human embryonic stem cell-derived cardiomyocytes. Stem Cell Res. 2010;4(3):189-200.
[11] GILLIE DJ, NOVICK SJ, DONOVAN BT, et al. Development of a high-throughput electrophysiological assay for the human ether-à-go-go related potassium channel hERG. J Pharmacol Toxicol Methods. 2013;67(1):33-44.
[12] COHEN JD, BABIARZ JE, ABRAMS RM, et al. Use of human stem cell derived cardiomyocytes to examine sunitinib mediated cardiotoxicity and electrophysiological alterations. Toxicol Appl Pharmacol. 2011;257(1):74-83.
[13] WHITMIRE ML, BRYAN P, HENRY TR, et al. Nonclinical dose formulation analysis method validation and sample analysis. AAPS J. 2010;12:628-634.
[14] ROCHE O, TRUBE G, ZUEGGE J, et al. A virtual screening method for prediction of the hERG potassium channel liability of compound libraries. Chembiochem. 2002;3(5):455-459.
[15] GINTANT GA, LIMBERIS JT, MCDERMOTT JS, et al. The canine Purkinje fiber: an in vitro model system for acquired long QT syndrome and drug-induced arrhythmogenesis. J Cardiovasc Pharmacol. 2001;37(5):607-618.
[16] TAKASUNA K, KATSUYOSHI C, MANABE S. Pre-clinical QT risk assessment in pharmaceutical companies-issues of current QT risk assessment. Biomol Ther. 2009;17(1):1-11.
[17] SHARMA A, BURRIDGE PW, MCKEITHAN WL, et al. High-throughput screening of tyrosine kinase inhibitor cardiotoxicity with human induced pluripotent stem cells. Sci Transl Med. 2017;9(377):eaaf2584.
[18] NAZEYROLLAS P, PREVOST A, BACCARD N, et al. Effects of amifostine on perfused isolated rat heart and on acute doxorubicin-induced cardiotoxicity. Cancer Chemother Pharmacol. 1999;43:227-232.
[19] TOKARSKA-SCHLATTNER M, ZAUGG M, DA SILVA R, et al. Acute toxicity of doxorubicin on isolated perfused heart:response of kinases regulating energy supply. Am J Physiol Heart Circ Physiol. 2005;289:H37-H47.
[20] WANG YX, KORTH M. Effects of doxorubicin on excitation-concentration coupling in guinea pig ventricular myocardium. Circ Res. 1995;76:645-653.
[21] STEINBERG JS, COHEN AJ, WASSERMAN AG, et al. Acute arrhythmogenicity of doxorubicin administration. Cancer. 1987;60:1213-1218.
[22] DUCROQ J, MOHA OU MAATI H, GUILBOT S, et al. Dexrazoxane protects the heart from acute doxorubicin-induced QT prolongation: a key role for IKs. Br J Pharmacol. 2010;159:93-101.
[23] FAIVRE S, DELBALDO C, VERA K, et al. Safety, pharmacokinetic, and antitumor activity of SU11248, a novel oral multitarget tyrosine kinase inhibitor, in patients with cancer. J Clin Oncol. 2006;24(1):25-35.
[24] KHOSRAVAN R, TOH M, LAFARGUE J, et al. Sunitinib pharmokinetic (PK) and safety data in subejcts with renal impairment and hemodialysis. 2008.
[25] CHU TF, RUPNICK MA, KERKELA R, et al. Cardiotoxicity associated with tyrosine kinase inhibitor sunitinib. Lancet. 2007;370:2011-2019.
[26] DI LORENZO G, AUTORINO R, BRUNI G, et al. Cardiovascular toxicity following sunitinib therapy in metastatic renal cell carcinoma: a multicenter analysis. Ann Oncol. 2009;20:1535-1542.
[27] COHEN JD, BABIARZ JE, ABRAMS RM, et al. Use of human stem cell derived cardiomyocytes to examine sunitinib mediated cardiotoxicity and electrophysiological alterations. Toxicol Appl Pharmacol. 2011;257(1):74-83.
[28] OBRIEN Z, F MOGHADDAM M. A systematic analysis of physicochemical and ADME properties of all small molecule kinase inhibitors approved by US FDA from January 2001 to October 2015. Curr Med Chem. 2017;24(29):3159-3184.
[29] KURATA Y, MIYAUCHI N, SUNO M, et al. Correlation of plasma crizotinib trough concentration with adverse events in patients with anaplastic lymphoma kinase positive non-small-cell lung cancer. J Pharm Health Care Sci. 2015;1(1):1-5.
[30] LESTER RM. Update on ICH E14/S7B cardiac safety regulations: the expanded role of preclinical assays and the “double-negative” scenario. Clin Pharmacol Drug Dev. 2021;10(9):964-973.
[31] CURIGLIANO G, SPITALERI G, DE BRAUD F, et al. QTc prolongation assessment in anticancer drug development: clinical and methodological issues. Ecancermedicalscience. 2009;3:130.
[32] PANG L, SAGER P, YANG X, et al. Workshop report: FDA workshop on improving cardiotoxicity assessment with human-relevant platforms. Circ Res. 2019;125(9):855-867.
[33] SHARMA A, BURRIDGE PW, MCKEITHAN WL, et al. High-throughput screening of tyrosine kinase inhibitor cardiotoxicity with human induced pluripotent stem cells. Sci Transl Med. 2017;9(377):eaaf2584.
[34] SELTZER JH, GINTANT G, AMIRI-KORDESTANI L, et al. Assessing cardiac safety in oncology drug development. Am Heart J. 2019;214:125-133.
[35] XIAO L, SALEM JE, CLAUSS S, et al. Ibrutinib-mediated atrial fibrillation attributable to inhibition of C-Terminal Src kinase. Circulation. 2020;142(25):2443-2455.