BACKGROUND: Anthracycline-induced heart failure has been traditionally attributed to direct iron-catalyzed oxidative damage. Dexrazoxane (DEX)-the only drug approved for its prevention-has been believed to protect the heart via its iron-chelating metabolite ADR-925. However, direct evidence is lacking, and recently proposed TOP2B (topoisomerase II beta) hypothesis challenged the original concept. METHODS: Pharmacokinetically guided study of the cardioprotective effects of clinically used DEX and its chelating metabolite ADR-925 (administered exogenously) was performed together with mechanistic experiments. The cardiotoxicity was induced by daunorubicin in neonatal ventricular cardiomyocytes in vitro and in a chronic rabbit model in vivo (n=50). RESULTS: Intracellular concentrations of ADR-925 in neonatal ventricular cardiomyocytes and rabbit hearts after treatment with exogenous ADR-925 were similar or exceeded those observed after treatment with the parent DEX. However, ADR-925 did not protect neonatal ventricular cardiomyocytes against anthracycline toxicity, whereas DEX exhibited significant protective effects (10-100 µmol/L; P<0.001). Unlike DEX, ADR-925 also had no significant impact on daunorubicin-induced mortality, blood congestion, and biochemical and functional markers of cardiac dysfunction in vivo (eg, end point left ventricular fractional shortening was 32.3±14.7%, 33.5±4.8%, 42.7±1.0%, and 41.5±1.1% for the daunorubicin, ADR-925 [120 mg/kg]+daunorubicin, DEX [60 mg/kg]+daunorubicin, and control groups, respectively; P<0.05). DEX, but not ADR-925, inhibited and depleted TOP2B and prevented daunorubicin-induced genotoxic damage. TOP2B dependency of the cardioprotective effects was probed and supported by experiments with diastereomers of a new DEX derivative. CONCLUSIONS: This study strongly supports a new mechanistic paradigm that attributes clinically effective cardioprotection against anthracycline cardiotoxicity to interactions with TOP2B but not metal chelation and protection against direct oxidative damage.
- Klíčová slova
- DNA topoisomerases, type II, anthracyclines, cardiotoxicity, dexrazoxane,
- MeSH
- antibiotika antitumorózní škodlivé účinky farmakologie MeSH
- antracykliny škodlivé účinky farmakologie MeSH
- daunomycin metabolismus farmakologie MeSH
- dexrazoxan škodlivé účinky farmakologie MeSH
- DNA-topoisomerasy typu II škodlivé účinky metabolismus MeSH
- inhibitory topoisomerasy II metabolismus MeSH
- kardiomyocyty účinky léků metabolismus MeSH
- kardiotoxicita farmakoterapie metabolismus prevence a kontrola MeSH
- lidé MeSH
- nemoci srdce farmakoterapie MeSH
- oxidační stres účinky léků MeSH
- srdeční selhání farmakoterapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antibiotika antitumorózní MeSH
- antracykliny MeSH
- daunomycin MeSH
- dexrazoxan MeSH
- DNA-topoisomerasy typu II MeSH
- inhibitory topoisomerasy II MeSH
The bisdioxopiperazine topoisomerase IIβ inhibitor ICRF-193 has been previously identified as a more potent analog of dexrazoxane (ICRF-187), a drug used in clinical practice against anthracycline cardiotoxicity. However, the poor aqueous solubility of ICRF-193 has precluded its further in vivo development as a cardioprotective agent. To overcome this issue, water-soluble prodrugs of ICRF-193 were prepared, their abilities to release ICRF-193 were investigated using a novel UHPLC-MS/MS assay, and their cytoprotective effects against anthracycline cardiotoxicity were tested in vitro in neonatal ventricular cardiomyocytes (NVCMs). Based on the obtained results, the bis(2-aminoacetoxymethyl)-type prodrug GK-667 was selected for advanced investigations due to its straightforward synthesis, sufficient solubility, low cytotoxicity and favorable ICRF-193 release. Upon administration of GK-667 to NVCMs, the released ICRF-193 penetrated well into the cells, reached sufficient intracellular concentrations and provided effective cytoprotection against anthracycline toxicity. The pharmacokinetics of the prodrug, ICRF-193 and its rings-opened metabolite was estimated in vivo after administration of GK-667 to rabbits. The plasma concentrations of ICRF-193 reached were found to be adequate to achieve cardioprotective effects in vivo. Hence, GK-667 was demonstrated to be a pharmaceutically acceptable prodrug of ICRF-193 and a promising drug candidate for further evaluation as a potential cardioprotectant against chronic anthracycline toxicity.
- MeSH
- antracykliny škodlivé účinky MeSH
- dexrazoxan chemie farmakologie MeSH
- diketopiperaziny chemie farmakologie MeSH
- DNA-topoisomerasy typu II metabolismus MeSH
- inhibitory topoisomerasy II chemie farmakologie MeSH
- kardiomyocyty účinky léků metabolismus MeSH
- kardiotonika chemie farmakologie MeSH
- kardiotoxicita farmakoterapie metabolismus MeSH
- králíci MeSH
- piperazin chemie farmakologie MeSH
- prekurzory léčiv chemie farmakologie MeSH
- razoxan chemie farmakologie MeSH
- voda chemie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 4,4'-(1,2-dimethyl-1,2-ethanediyl)bis-2,6-piperazinedione MeSH Prohlížeč
- antracykliny MeSH
- dexrazoxan MeSH
- diketopiperaziny MeSH
- DNA-topoisomerasy typu II MeSH
- inhibitory topoisomerasy II MeSH
- kardiotonika MeSH
- piperazin MeSH
- prekurzory léčiv MeSH
- razoxan MeSH
- voda MeSH
Bisdioxopiperazine agent dexrazoxane (ICRF-187) has been the only effective and approved drug for prevention of chronic anthracycline cardiotoxicity. However, the structure-activity relationships (SARs) of its cardioprotective effects remain obscure owing to limited investigation of its derivatives/analogs and uncertainties about its mechanism of action. To fill these knowledge gaps, we tested the hypothesis that dexrazoxane derivatives exert cardioprotection via metal chelation and/or modulation of topoisomerase IIβ (Top2B) activity in chronic anthracycline cardiotoxicity. Dexrazoxane was alkylated in positions that should not interfere with the metal-chelating mechanism of cardioprotective action; that is, on dioxopiperazine imides or directly on the dioxopiperazine ring. The protective effects of these agents were assessed in vitro in neonatal cardiomyocytes. All studied modifications of dexrazoxane molecule, including simple methylation, were found to abolish the cardioprotective effects. Because this challenged the prevailing mechanistic concept and previously reported data, the two closest derivatives [(±)-4,4'-(propane-1,2-diyl)bis(1-methylpiperazine-2,6-dione) and 4-(2-(3,5-dioxopiperazin-1-yl)ethyl)-3-methylpiperazine-2,6-dione] were thoroughly scrutinized in vivo using a rabbit model of chronic anthracycline cardiotoxicity. In contrast to dexrazoxane, both compounds failed to protect the heart, as demonstrated by mortality, cardiac dysfunction, and myocardial damage parameters, although the pharmacokinetics and metal-chelating properties of their metabolites were comparable to those of dexrazoxane. The loss of cardiac protection was shown to correlate with their abated potential to inhibit and deplete Top2B both in vitro and in vivo. These findings suggest a very tight SAR between bisdioxopiperazine derivatives and their cardioprotective effects and support Top2B as a pivotal upstream druggable target for effective cardioprotection against anthracycline cardiotoxicity. SIGNIFICANCE STATEMENT: This study has revealed the previously unexpected tight structure-activity relationships of cardioprotective effects in derivatives of dexrazoxane, which is the only drug approved for the prevention of cardiomyopathy and heart failure induced by anthracycline anticancer drugs. The data presented in this study also strongly argue against the importance of metal-chelating mechanisms for the induction of this effect and support the viability of topoisomerase IIβ as an upstream druggable target for effective and clinically translatable cardioprotection.
- MeSH
- antracykliny škodlivé účinky MeSH
- dexrazoxan farmakologie MeSH
- DNA-topoisomerasy typu II metabolismus MeSH
- HL-60 buňky MeSH
- inhibitory topoisomerasy II farmakologie MeSH
- kardiomyocyty účinky léků metabolismus MeSH
- kardiomyopatie farmakoterapie metabolismus MeSH
- kardiotoxicita farmakoterapie MeSH
- králíci MeSH
- krysa rodu Rattus MeSH
- lidé MeSH
- modely u zvířat MeSH
- myokard metabolismus MeSH
- nádorové buněčné linie MeSH
- ochranné látky farmakologie MeSH
- potkani Wistar MeSH
- srdce účinky léků MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- krysa rodu Rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antracykliny MeSH
- dexrazoxan MeSH
- DNA-topoisomerasy typu II MeSH
- inhibitory topoisomerasy II MeSH
- ochranné látky MeSH
Novel dexrazoxane derivative JR-311 was prepared to investigate structure-activity relationships and mechanism(s) of protection against anthracycline cardiotoxicity. Its cardioprotective, antiproliferative, iron (Fe) chelation and inhibitory and/or depletory activities on topoisomerase IIbeta (TOP2B) were examined and compared with dexrazoxane. While in standard assay, JR-311 failed in both cardioprotection and depletion of TOP2B, its repeated administration to cell culture media led to depletion of TOP2B and significant protection of isolated rat neonatal ventricular cardiomyocytes from daunorubicin-induced damage. This effect was explained by a focused analytical investigation that revealed rapid JR-311 decomposition, resulting in negligible intracellular concentrations of the parent compound but high exposure of cells to the decomposition products, including Fe-chelating JR-H2. Although chemical instability is an obstacle for the development of JR-311, this study identified a novel dexrazoxane analogue with preserved pharmacodynamic properties, contributed to the investigation of structure-activity relationships and suggested that the cardioprotection of bis-dioxopiperazines is likely attributed to TOP2B activity of the parent compound rather than Fe chelation of their hydrolytic metabolites/degradation products. Moreover, this study highlights the importance of early stability testing during future development of novel dexrazoxane analogues.
- Klíčová slova
- Bis-dioxopiperazine, Cardioprotection, JR-311, Structure-activity relationship, Topoisomerase,
- MeSH
- antracykliny toxicita MeSH
- chelátory železa farmakologie MeSH
- daunomycin toxicita MeSH
- dexrazoxan analogy a deriváty farmakologie MeSH
- diketopiperaziny farmakologie MeSH
- DNA-topoisomerasy typu II metabolismus MeSH
- kardiomyocyty účinky léků MeSH
- kardiotonika farmakologie MeSH
- kardiotoxicita farmakoterapie etiologie MeSH
- krysa rodu Rattus MeSH
- kultivované buňky MeSH
- novorozená zvířata MeSH
- potkani Wistar MeSH
- proliferace buněk účinky léků MeSH
- vztahy mezi strukturou a aktivitou MeSH
- železo metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 2,5-dioxopiperazine MeSH Prohlížeč
- antracykliny MeSH
- chelátory železa MeSH
- daunomycin MeSH
- dexrazoxan MeSH
- diketopiperaziny MeSH
- DNA-topoisomerasy typu II MeSH
- JR-311 MeSH Prohlížeč
- kardiotonika MeSH
- železo MeSH
Dexrazoxane (DEX) is a clinically available cardioprotectant that reduces the toxicity induced by anthracycline (ANT) anticancer drugs; however, DEX is seldom used and its action is poorly understood. Inorganic nitrate/nitrite has shown promising results in myocardial ischemia-reperfusion injury and recently in acute high-dose ANT cardiotoxicity. However, the utility of this approach for overcoming clinically more relevant chronic forms of cardiotoxicity remains elusive. Hence, in this study, the protective potential of inorganic nitrate and nitrite against chronic ANT cardiotoxicity was investigated, and the results were compared to those using DEX. Chronic cardiotoxicity was induced in rabbits with daunorubicin (DAU). Sodium nitrate (1g/L) was administered daily in drinking water, while sodium nitrite (0.15 or 5mg/kg) or DEX (60mg/kg) was administered parenterally before each DAU dose. Although oral nitrate induced a marked increase in plasma NOx, it showed no improvement in DAU-induced mortality, myocardial damage or heart failure. Instead, the higher nitrite dose reduced the incidence of end-stage cardiotoxicity, prevented related premature deaths and significantly ameliorated several molecular and cellular perturbations induced by DAU, particularly those concerning mitochondria. The latter result was also confirmed in vitro. Nevertheless, inorganic nitrite failed to prevent DAU-induced cardiac dysfunction and molecular remodeling in vivo and failed to overcome the cytotoxicity of DAU to cardiomyocytes in vitro. In contrast, DEX completely prevented all of the investigated molecular, cellular and functional perturbations that were induced by DAU. Our data suggest that the difference in cardioprotective efficacy between DEX and inorganic nitrite may be related to their different abilities to address a recently proposed upstream target for ANT in the heart - topoisomerase IIβ.
- Klíčová slova
- Anthracyclines, Cardioprotection, Cardiotoxicity, Dexrazoxane, Inorganic nitrate and nitrite,
- MeSH
- antibiotika antitumorózní škodlivé účinky MeSH
- daunomycin škodlivé účinky MeSH
- dexrazoxan farmakologie MeSH
- DNA vazebné proteiny antagonisté a inhibitory metabolismus MeSH
- DNA-topoisomerasy typu II metabolismus MeSH
- dusičnany farmakologie MeSH
- dusitan sodný farmakologie MeSH
- intravenózní infuze MeSH
- kardiomyocyty metabolismus patologie MeSH
- kardiotonika farmakologie MeSH
- kardiotoxicita metabolismus patologie prevence a kontrola MeSH
- králíci MeSH
- myokard metabolismus patologie MeSH
- rozvrh dávkování léků MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- antibiotika antitumorózní MeSH
- daunomycin MeSH
- dexrazoxan MeSH
- DNA vazebné proteiny MeSH
- DNA-topoisomerasy typu II MeSH
- dusičnany MeSH
- dusitan sodný MeSH
- kardiotonika MeSH
- sodium nitrate MeSH Prohlížeč
Anthracyclines (such as doxorubicin or daunorubicin) are among the most effective anticancer drugs, but their usefulness is hampered by the risk of irreversible cardiotoxicity. Dexrazoxane (ICRF-187) is the only clinically approved cardioprotective agent against anthracycline cardiotoxicity. Its activity has traditionally been attributed to the iron-chelating effects of its metabolite with subsequent protection from oxidative stress. However, dexrazoxane is also a catalytic inhibitor of topoisomerase II (TOP2). Therefore, we examined whether dexrazoxane and two other TOP2 catalytic inhibitors, namely sobuzoxane (MST-16) and merbarone, protect cardiomyocytes from anthracycline toxicity and assessed their effects on anthracycline antineoplastic efficacy. Dexrazoxane and two other TOP2 inhibitors protected isolated neonatal rat cardiomyocytes against toxicity induced by both doxorubicin and daunorubicin. However, none of the TOP2 inhibitors significantly protected cardiomyocytes in a model of hydrogen peroxide-induced oxidative injury. In contrast, the catalytic inhibitors did not compromise the antiproliferative effects of the anthracyclines in the HL-60 leukemic cell line; instead, synergistic interactions were mostly observed. Additionally, anthracycline-induced caspase activation was differentially modulated by the TOP2 inhibitors in cardiac and cancer cells. Whereas dexrazoxane was upon hydrolysis able to significantly chelate intracellular labile iron ions, no such effect was noted for either sobuzoxane or merbarone. In conclusion, our data indicate that dexrazoxane may protect cardiomyocytes via its catalytic TOP2 inhibitory activity rather than iron-chelation activity. The differential expression and/or regulation of TOP2 isoforms in cardiac and cancer cells by catalytic inhibitors may be responsible for the selective modulation of anthracycline action observed.
- MeSH
- antracykliny farmakologie MeSH
- biokatalýza účinky léků MeSH
- buněčný cyklus účinky léků MeSH
- daunomycin farmakologie MeSH
- dexrazoxan farmakologie MeSH
- DNA-topoisomerasy typu II metabolismus MeSH
- doxorubicin farmakologie MeSH
- glutathion metabolismus MeSH
- glutathiondisulfid metabolismus MeSH
- HL-60 buňky MeSH
- inhibitory topoisomerasy II farmakologie MeSH
- kardiomyocyty cytologie účinky léků metabolismus MeSH
- kaspasy metabolismus MeSH
- krysa rodu Rattus MeSH
- kultivované buňky MeSH
- lékové interakce MeSH
- lidé MeSH
- novorozená zvířata MeSH
- piperaziny farmakologie MeSH
- potkani Wistar MeSH
- proliferace buněk účinky léků MeSH
- průtoková cytometrie MeSH
- thiobarbituráty farmakologie MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antracykliny MeSH
- daunomycin MeSH
- dexrazoxan MeSH
- DNA-topoisomerasy typu II MeSH
- doxorubicin MeSH
- glutathion MeSH
- glutathiondisulfid MeSH
- inhibitory topoisomerasy II MeSH
- kaspasy MeSH
- merbarone MeSH Prohlížeč
- piperaziny MeSH
- sobuzoxane MeSH Prohlížeč
- thiobarbituráty MeSH
