Alisertib (MLN8237), a novel Aurora A kinase inhibitor, is currently being clinically tested in late-phase trials for the therapy of various malignancies. In the present work, we describe alisertib's potential to perpetrate pharmacokinetic drug-drug interactions (DDIs) and/or to act as an antagonist of multidrug resistance (MDR). In accumulation assays, alisertib potently inhibited ABCC1 transporter, but not ABCB1 or ABCG2. The results of molecular modeling suggested a bifunctional mechanism for interaction on ABCC1. In addition, alisertib was characterized as a low- to moderate-affinity inhibitor of recombinant CYP3A4, CYP2C8, CYP2C9, CYP2C19, and CYP2D6 isoenzymes, but without potential clinical relevance. Drug combination studies revealed the capability of alisertib to synergistically antagonize ABCC1-mediated resistance to daunorubicin. Although alisertib exhibited substrate characteristics toward ABCB1 transporter in monolayer transport assays, comparative proliferation studies showed lack of its MDR-victim behavior in cells overexpressing ABCB1 as well as ABCG2 and ABCC1. Lastly, alisertib did not affect the expression of ABCC1, ABCG2, ABCB1 transporters and CYP1A2, CYP3A4, CYP2B6 isozymes on mRNA level in various systemic and tumoral models. In conclusion, our study suggests that alisertib is a drug candidate with negligible potential for perpetrating systemic pharmacokinetic DDIs on ABCB1, ABCG2 and cytochromes P450. In addition, we introduce alisertib as an effective dual-activity chemosensitizer whose MDR-antagonistic capacities are not impaired by efflux or effect on MDR phenotype. Our in vitro findings provide important pieces of information for clinicians when introducing alisertib into the clinical area.
- MeSH
- ABC transportér z rodiny G, člen 2 genetika metabolismus MeSH
- azepiny farmakokinetika farmakologie MeSH
- buněčné linie MeSH
- katalytická doména MeSH
- konformace proteinů MeSH
- lékové interakce MeSH
- lidé MeSH
- molekulární modely MeSH
- P-glykoprotein genetika metabolismus MeSH
- proteiny spojené s mnohočetnou rezistencí k lékům antagonisté a inhibitory MeSH
- psi MeSH
- pyrimidiny farmakokinetika farmakologie MeSH
- regulace genové exprese účinky léků MeSH
- simulace molekulového dockingu MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Talazoparib (Talzenna) is a novel poly (adenosine diphosphate-ribose) polymerase (PARP) inhibitor that is clinically used for the therapy of breast cancer. Furthermore, the drug has shown antitumor activity against different cancer types, including non-small cell lung cancer (NSCLC). In this work, we investigated the possible inhibitory interactions of talazoparib toward selected ATP-binding cassette (ABC) drug efflux transporters and cytochrome P450 biotransformation enzymes (CYPs) and evaluated its position in multidrug resistance (MDR). In accumulation studies, talazoparib interacted with the ABCC1 and ABCG2 transporters, but there were no significant effects on ABCB1. Furthermore, incubation assays revealed a negligible capacity of the tested drug to inhibit clinically relevant CYPs. In in vitro drug combination experiments, talazoparib synergistically reversed daunorubicin and mitoxantrone resistance in cells with ABCC1 and ABCG2 expression, respectively. Importantly, the position of an effective MDR modulator was further confirmed in drug combinations performed in ex vivo NSCLC patients-derived explants, whereas the possible victim role was refuted in comparative proliferation experiments. In addition, talazoparib had no significant effects on the mRNA-level expressions of MDR-related ABC transporters in the MCF-7 cellular model. In summary, our study presents a comprehensive overview on the pharmacokinetic drug-drug interactions (DDI) profile of talazoparib. Moreover, we introduced talazoparib as an efficient MDR antagonist.
- MeSH
- ABC transportér z rodiny G, člen 2 genetika MeSH
- ABC transportéry genetika metabolismus MeSH
- lidé MeSH
- mnohočetná léková rezistence MeSH
- nádorové proteiny metabolismus MeSH
- nádory plic * MeSH
- nemalobuněčný karcinom plic * farmakoterapie genetika MeSH
- P-glykoproteiny genetika MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Tepotinib is a novel tyrosine kinase inhibitor recently approved for the treatment of non-small cell lung cancer (NSCLC). In this study, we evaluated the tepotinib's potential to perpetrate pharmacokinetic drug interactions and modulate multidrug resistance (MDR). Accumulation studies showed that tepotinib potently inhibits ABCB1 and ABCG2 efflux transporters, which was confirmed by molecular docking. In addition, tepotinib inhibited several recombinant cytochrome P450 (CYP) isoforms with varying potency. In subsequent drug combination experiments, tepotinib synergistically reversed daunorubicin and mitoxantrone resistance in cells with ABCB1 and ABCG2 overexpression, respectively. Remarkably, MDR-modulatory properties were confirmed in ex vivo explants derived from NSCLC patients. Furthermore, we demonstrated that anticancer effect of tepotinib is not influenced by the presence of ABC transporters associated with MDR, although monolayer transport assays designated it as ABCB1 substrate. Finally, tested drug was observed to have negligible effect on the expression of clinically relevant drug efflux transporters and CYP enzymes. In conclusion, our findings provide complex overview on the tepotinib's drug interaction profile and suggest a promising novel therapeutic strategy for future clinical investigations.
- MeSH
- ABC transportéry antagonisté a inhibitory MeSH
- antitumorózní látky farmakologie MeSH
- chemorezistence účinky léků MeSH
- cytostatické látky farmakologie MeSH
- lékové interakce * MeSH
- lidé MeSH
- mnohočetná léková rezistence účinky léků MeSH
- nádory plic farmakoterapie metabolismus patologie MeSH
- nemalobuněčný karcinom plic farmakoterapie metabolismus patologie MeSH
- piperidiny farmakologie MeSH
- pyridaziny farmakologie MeSH
- pyrimidiny farmakologie MeSH
- techniky in vitro MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Olaparib is a potent poly (ADP-ribose) polymerase inhibitor currently used in targeted therapy for treating cancer cells with BRCA mutations. Here we investigate the possible interference of olaparib with daunorubicin (Daun) metabolism, mediated by carbonyl-reducing enzymes (CREs), which play a significant role in the resistance of cancer cells to anthracyclines. Incubation experiments with the most active recombinant CREs showed that olaparib is a potent inhibitor of the aldo-keto reductase 1C3 (AKR1C3) enzyme. Subsequent inhibitory assays in the AKR1C3-overexpressing cellular model transfected human colorectal carcinoma HCT116 cells, demonstrating that olaparib significantly inhibits AKR1C3 at the intracellular level. Consequently, molecular docking studies have supported these findings and identified the possible molecular background of the interaction. Drug combination experiments in HCT116, human liver carcinoma HepG2, and leukemic KG1α cell lines showed that this observed interaction can be exploited for the synergistic enhancement of Daun's antiproliferative effect. Finally, we showed that olaparib had no significant effect on the mRNA expression of AKR1C3 in HepG2 and KG1α cells. In conclusion, our data demonstrate that olaparib interferes with anthracycline metabolism, and suggest that this phenomenon might be utilized for combating anthracycline resistance.
- Publikační typ
- časopisecké články MeSH
The anthelmintic drug ivermectin (IVM), used frequently especially in veterinary medicine, enters the environment mainly via excrements in pastures and could negatively affect non-target organisms including plants. The present study was designed to follow up on our previous investigations into IVM metabolism and its effects in the common meadow plant ribwort plantain (Plantago lanceolata L.) during long-term exposure of both cell suspensions and whole plant regenerants. IVM uptake, distribution, and biotransformation pathways were studied using UHPLC-MS analysis. In addition, the IVM effect on antioxidant enzymes activities, proline concentration, the content of all polyphenols, and the level of the main bioactive secondary metabolites was also tested with the goal of learning more about IVM-induced stress in the plant organism. Our results showed that the ribwort plantain was able to uptake IVM and transform it via demethylation and hydroxylation. Seven and six metabolites respectively were detected in cell suspensions and in the roots of regenerants. However, only the parent drug IVM was detected in the leaves of the regenerants. IVM accumulated in the roots and leaves of plants might negatively affect ecosystems due to its toxicity to herbivorous invertebrates. As IVM exposition increased the activity of catalase, the concentration of proline and polyphenols, as well as decreased the activity of ascorbate peroxidase and the concentration of the bioactive compounds acteoside and aucubin, long-term exposition of the ribwort plantain to IVM caused abiotic stress and might decrease the medicinal value of this herb.
- MeSH
- ekosystém MeSH
- ivermektin MeSH
- ovoce MeSH
- Plantago * MeSH
- zelenina MeSH
- Publikační typ
- časopisecké články MeSH
Entrectinib is a new tyrosine kinase inhibitor that was recently approved for the treatment of ROS1-positive metastatic non-small cell lung cancer (NSCLC). In this study, we aimed to characterize its potential to act as a modulator of pharmacokinetic cytostatic resistance and perpetrator of drug interactions. In accumulation studies, entrectinib exhibited potent inhibition of ABCB1, while only moderate interaction was recorded for ABCG2 and ABCC1 efflux transporters. Furthermore, incubation assays revealed the potential of this drug to inhibit various recombinant cytochrome P450 enzymes, which can be ranked according to inhibitory affinities as follows: CYP2C8 ≈ CYP3A4 > CYP2C9 > CYP2C19 ≈ CYP3A5 > CYP2D6 > CYP2B6 > CYP1A2. Additionally, in silico docking analysis confirmed entrectinib's interactions with ABCB1 and CYP3A4 and resolved their possible molecular background. In subsequent drug combination experiments, we demonstrated the ability of entrectinib to synergize with daunorubicin in various ABCB1-expressing cellular models. Moreover, the comparative proliferation study results suggested that the anticancer efficacy of entrectinib is not affected by the functional presence of tested ABC transporters. In contrast to ABCB1-related data, no resistance reversal effect was recorded for the combination with docetaxel in HepG2-CYP3A4 cells. In the final experimental set, we observed no significant changes in ABCB1, ABCG2, ABCC1 or CYP3A4 gene expression in NSCLC cells exposed to entrectinib. In summary, our work indicates that entrectinib may be a perpetrator of clinically relevant pharmacokinetic drug interactions and modulator of ABCB1-mediated resistance. Our in vitro results might provide a valuable foundation for future clinical investigations.
- MeSH
- benzamidy farmakologie MeSH
- buňky A549 MeSH
- buňky Hep G2 MeSH
- buňky MDCK MeSH
- chemorezistence účinky léků fyziologie MeSH
- cytochrom P-450 CYP3A * chemie metabolismus MeSH
- cytostatické látky farmakologie MeSH
- indazoly farmakologie MeSH
- inhibitory proteinkinas farmakologie MeSH
- lidé MeSH
- mnohočetná léková rezistence účinky léků fyziologie MeSH
- nádorové buněčné linie MeSH
- P-glykoproteiny antagonisté a inhibitory chemie metabolismus MeSH
- psi MeSH
- sekundární struktura proteinů MeSH
- simulace molekulového dockingu metody MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Ensartinib (X-396) is a promising tyrosine kinase inhibitor currently undergoing advanced clinical evaluation for the treatment of non-small cell lung cancer. In this work, we investigate possible interactions of this promising drug candidate with ATP-binding cassette (ABC) drug efflux transporters and cytochrome P450 biotransformation enzymes (CYPs), which play major roles in multidrug resistance (MDR) and pharmacokinetic drug-drug interactions (DDIs). Accumulation studies showed that ensartinib is a potent inhibitor of ABCB1 and ABCG2 transporters. Additionally, incubation experiments with recombinant CYPs showed that ensartinib significantly inhibits CYP3A4 and CYP2C9. Subsequent molecular docking studies confirmed these findings. Drug combination experiments demonstrated that ensartinib synergistically potentiates the antiproliferative effects of daunorubicin, mitoxantrone, and docetaxel in ABCB1, ABCG2, and CYP3A4-overexpressing cellular models, respectively. Advantageously, ensartinib's antitumor efficiency was not compromised by the presence of MDR-associated ABC transporters, although it acted as a substrate of ABCB1 in Madin-Darby Canine Kidney II (MDCKII) monolayer transport assays. Finally, we demonstrated that ensartinib had no significant effect on the mRNA-level expression of examined transporters and enzymes in physiological and lung tumor cellular models. In conclusion, ensartinib may perpetrate clinically relevant pharmacokinetic DDIs and modulate ABCB1-, ABCG2-, and CYP3A4-mediated MDR. The in vitro findings presented here will provide a valuable foundation for future in vivo investigations.
- Publikační typ
- časopisecké články MeSH
Fenbendazole, a broad spectrum anthelmintic used especially in veterinary medicine, may impact non-target organisms in the environment. Nevertheless, information about the effects of fenbendazole in plants is limited. We investigated the biotransformation of fenbendazole and the effect of fenbendazole and its metabolites on gene expression in the model plant Arabidopsis thaliana. High-sensitive UHPLC coupled with tandem mass spectrometry, RNA-microarray analysis together with qPCR verification and nanoLC-MS proteome analysis were used in this study. Twelve fenbendazole metabolites were identified in the roots and leaves of A. thaliana plants. Hydroxylation, S-oxidation and glycosylation represent the main fenbendazole biotransformation pathways. Exposure of A. thaliana plants to 5 μM fenbendazole for 24 and 72 h significantly affected gene and protein expression. The changes in transcriptome were more pronounced in the leaves than in roots, protein expression was more greatly affected in the roots at a shorter period of exposure (24 h) and in leaf rosettes over a longer period (72 h). Up-regulated (>2-fold change, p < 0.1) proteins are involved in various biological processes (electron transport, energy generating pathways, signal transduction, transport), and in response to stresses (e.g. catalase, superoxide dismutase, cytochromes P450, UDP-glycosyltransferases). Some of the proteins which were up-regulated after fenbendazole-exposure probably participate in fenbendazole biotransformation (e.g. cytochromes P450, UDP-glucosyltransferases). Finally, fenbendazole in plants significantly affects many physiological and metabolic processes and thus the contamination of ecosystems by manure containing this anthelmintic should be restricted.
- MeSH
- anthelmintika metabolismus MeSH
- Arabidopsis účinky léků metabolismus MeSH
- fenbendazol metabolismus farmakokinetika MeSH
- kořeny rostlin metabolismus MeSH
- listy rostlin metabolismus MeSH
- proteiny huseníčku účinky léků metabolismus MeSH
- proteom účinky léků metabolismus MeSH
- proteomika metody MeSH
- regulace genové exprese u rostlin MeSH
- transkriptom účinky léků MeSH
- Publikační typ
- časopisecké články MeSH
Drugs are potentially dangerous environmental contaminants, as they are designed to have biological effects at low concentrations. Monepantel (MOP), an amino-acetonitrile derivative, is frequently used veterinary anthelmintics, but information about MOP environmental circulation and impact is almost non-existent. We studied the phytotoxicity, uptake and biotransformation of MOP in two fodder plants, Plantago lanceolata and Medicago sativa. The seeds and whole plant regenerants were cultivated with MOP. The plant roots and the leaves were collected after 1, 2, 3, 4, 5 and 6 weeks of cultivation. The lengths of roots and proline concentrations in the roots and leaves were measured to evaluate MOP phytotoxicity. The UHPLC-MS/MS technique with a Q-TOF mass analyser was used for the identification and semi-quantification of MOP and its metabolites. Our results showed no phytotoxicity of MOP. However, both plants were able to uptake, transport and metabolize MOP. Comparing both plants, the uptake of MOP was much more extensive in Medicago sativa (almost 10-times) than in Plantago lanceolate. Moreover, 9 various metabolites of MOP were detected in Medicago sativa, while only 7 MOP metabolites were found in Plantago lanceolata. Based on metabolites structures, scheme of the metabolic pathways of MOP in both plants was proposed. MOP and its main metabolite (MOP sulfone), both anthelmintically active, were present not only in roots but also in leaves that can be consumed by animals. This indicates the potential for undesirable circulation of MOP in the environment, which could lead to many pharmacological and toxicological consequences.
- MeSH
- aminoacetonitrily analogy a deriváty farmakokinetika toxicita MeSH
- anthelmintika toxicita MeSH
- biologický transport MeSH
- biotransformace MeSH
- dobytek MeSH
- krmivo pro zvířata toxicita MeSH
- Medicago sativa metabolismus MeSH
- metabolické sítě a dráhy MeSH
- pastviny * MeSH
- Plantago metabolismus MeSH
- sulfony MeSH
- tandemová hmotnostní spektrometrie MeSH
- znečištění životního prostředí * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Veterinary drugs enter the environment in many ways and may affect non-target organisms, including plants. The present project was focused on the biotransformation of ivermectin (IVM), one of the mostly used anthelmintics, in the model plant Arabidopsis thaliana. Our results certified the ability of plants to uptake IVM by roots and translocate it to the aboveground parts. Using UHPLC-MS/MS, six metabolites in roots and only the parent drug in rosettes were found after 24- and 72-h incubation of A. thaliana with IVM. The metabolites were formed only via hydroxylation and demethylation, with no IVM conjugates detected. Although IVM did not induce changes in the activity of antioxidant enzymes in A. thaliana rosettes, the expression of genes was significantly affected. Surprisingly, a higher number of transcripts, 300 and 438, respectively, was dysregulated in the rosettes than in roots. The significantly affected genes play role in response to salt, osmotic and water deprivation stress, in response to pathogens and in ion homeostasis. We hypothesize that the above described changes in gene transcription in A. thaliana resulted from disrupted ionic homeostasis caused by certain ionophore properties of IVM. Our results underlined the negative impact of IVM presence in the environment.
- MeSH
- anthelmintika metabolismus farmakokinetika MeSH
- Arabidopsis účinky léků genetika metabolismus MeSH
- biotransformace MeSH
- ivermektin metabolismus farmakokinetika MeSH
- kořeny rostlin účinky léků metabolismus MeSH
- tandemová hmotnostní spektrometrie MeSH
- transkriptom účinky léků MeSH
- Publikační typ
- časopisecké články MeSH
