ABSTRACT
Apparently, both a decrease in beta cell function and in beta cell mass contribute to the progressive worsening of type 2 diabetes. So, it is of particular interest to define factors which are relevant for the regulation of insulin secretion and at the same time for the maintenance of beta cell mass. The NADPH-thioredoxin system has a candidate role for such a dual function. Here, we have characterized the effects of a highly specific inhibitor of thioredoxin reductase, AM12, on the viability and function of insulin-secreting MIN6 cells and isolated NMRI mouse islets. Viability was checked by MTT testing and the fluorescent live-dead assay. Apoptosis was assessed by annexin V assay. Insulin secretion of perifused islets was measured by ELISA. The cytosolic Ca2+ concentration was measured by the Fura technique. Acute exposure of perifused pancreatic islets to 5 µM AM12 was without significant effect on insulin secretion. Islets cultured for 24 h in 0.5 or 5 µM AM12 showed unchanged basal secretion during perifusion, but the response to 30 mM glucose was significantly enhanced by 5 µM. Twenty-four-hour exposure to 5 µM AM12 proved to be without effect on the viability of MIN6 cells, whereas longer exposure was clearly toxic. Islets were more susceptible, showing initial signs of apoptosis after 24-h exposure to 5 µM AM12. The activity of the NADPH-thioredoxin system is indispensable for beta cell viability but may have a limiting effect on glucose-induced insulin secretion.
Subject(s)
Insulin Secretion/drug effects , Insulin-Secreting Cells/drug effects , Islets of Langerhans/drug effects , Thioredoxin-Disulfide Reductase/antagonists & inhibitors , Animals , Apoptosis/drug effects , Benzene Derivatives/pharmacology , Cell Line , Cell Survival/drug effects , Dose-Response Relationship, Drug , Female , Glucose/metabolism , Insulin-Secreting Cells/metabolism , Islets of Langerhans/metabolism , Mice , Organogold Compounds/pharmacology , Thioredoxin-Disulfide Reductase/metabolism , Time FactorsABSTRACT
Gold alkynyl complexes with phosphane ligands of the type (alkynyl)Au(I)(phosphane) represent a group of bioorganometallics, which has only recently been evaluated biologically in more detail. Structure-activity-relationship studies regarding the residues of the phosphane ligand (P(Ph)3, P(2-furyl)3, P(DAPTA)3, P(PTA)3, P(Et)3, P(Me)3) of complexes with an 4-ethynylanisole alkyne ligand revealed no strong differences concerning cytotoxicity. However, a relevant preference for the heteroatom free alkyl/aryl residues concerning inhibition of the target enzyme thioredoxin reductase was evident. Complex 1 with the triphenylphosphane ligand was selected for further studies, in which clear effects on cell morphology were monitored by time-lapse microscopy. Effects on cellular signaling were determined by ELISA microarrays and showed a significant induction of the phosphorylation of ERK1 (extracellular signal related kinase 1), ERK2 and HSP27 (heat shock protein 27) in HT-29 cells. Application of 1 in-vivo in a mouse xenograft model was found to be challenging due to the low solubility of the complex and required a formulation strategy based on a peanut oil nanoemulsion.
Subject(s)
Antineoplastic Agents/chemical synthesis , Coordination Complexes/chemical synthesis , Gold/chemistry , Organogold Compounds/chemical synthesis , Phosphines/chemistry , Animals , Anisoles/chemistry , Antineoplastic Agents/pharmacology , Cations, Monovalent , Cell Proliferation/drug effects , Cell Survival/drug effects , Coordination Complexes/pharmacology , Female , HSP27 Heat-Shock Proteins/genetics , HSP27 Heat-Shock Proteins/metabolism , HT29 Cells , Heat-Shock Proteins , Humans , Ligands , Lung Neoplasms/drug therapy , Lung Neoplasms/pathology , Mice , Mice, Inbred BALB C , Mice, Nude , Mitogen-Activated Protein Kinase 1/genetics , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/genetics , Mitogen-Activated Protein Kinase 3/metabolism , Molecular Chaperones , Organogold Compounds/pharmacology , Structure-Activity Relationship , Thioredoxin-Disulfide Reductase/antagonists & inhibitors , Thioredoxin-Disulfide Reductase/genetics , Thioredoxin-Disulfide Reductase/metabolism , Xenograft Model Antitumor AssaysABSTRACT
Di-n-butyltin(IV) carboxylate and tri-n-butyltin(IV) carboxylate derivatives have demonstrated strong cytotoxic effects in different types of tumor cells. Complexes with carboxylate ligands that contain maleimide and naphthalimide derived partial structures were synthesized, characterized and investigated for inhibition of the tumor-relevant enzyme thioredoxin reductase and antiproliferative effects in cancer cells. The complexes were moderate inhibitors of thioredoxin reductase with activities in the micromolar range and triggered strong cytotoxic effects in MCF-7 breast cancer and HT-29 colon carcinoma cells. Interestingly, selected complexes were highly active in vincristine and daunorubicin resistant Nalm-6 cells.
Subject(s)
Carboxylic Acids/metabolism , Maleimides/chemistry , Naphthalimides/chemistry , Organotin Compounds/pharmacology , Thioredoxin-Disulfide Reductase/antagonists & inhibitors , Cell Line, Tumor , Humans , Ligands , Magnetic Resonance Spectroscopy , Organotin Compounds/chemistry , Spectrometry, Mass, Electrospray IonizationABSTRACT
The synthesis and characterization of three propynyloxycoumarins are reported in this work together with the formation of three different series of gold(i) organometallic complexes. Neutral complexes are constituted by water soluble phosphines (PTA and DAPTA) which confer water solubility to them. The X-ray crystal structure of 7-(prop-2-in-1-yloxy)-1-benzopyran-2-one and its corresponding dialkynyl complex is also shown and the formation of rectangular dimers for the gold derivative in the solid state can be observed. A detailed analysis of the absorption and emission spectra of both ligands and complexes allows us to attribute the luminescent behaviour to the coumarin organic ligand. Moreover, the presence of the gold(i) metal atom seems to be responsible for an increase of coumarin phosphorescence emission. The biological activity of the complexes showed that the anionic complexes triggered strong cytotoxic effects in two different cell lines whereas the neutral gold alkynyl complexes led to lower effects against tumor cell growth. Thioredoxin reductase (TrxR) inhibition was very strong in the case of the neutral complexes (IC50 values below 0.1 µM) but moderate for the anionic complexes (IC50 values above 0.8 µM).
Subject(s)
Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Coumarins/chemistry , Coumarins/pharmacology , Organogold Compounds/chemistry , Organogold Compounds/pharmacology , Antineoplastic Agents/chemical synthesis , Cell Line, Tumor , Coumarins/chemical synthesis , Crystallography, X-Ray , Drug Screening Assays, Antitumor , Humans , Luminescence , Models, Molecular , Neoplasms/drug therapy , Organogold Compounds/chemical synthesis , Phosphines/chemical synthesis , Phosphines/chemistry , Phosphines/pharmacology , Solubility , Thioredoxin-Disulfide Reductase/antagonists & inhibitors , Water/chemistryABSTRACT
Thioredoxin reductase (TrxR) is overexpressed in cancer cells and is therefore a putative cancer target. Inhibition of this enzyme is considered an important strategy for the development of new chemotherapeutic agents with a specific mechanism of action. Organotin compounds have been described as experimental antitumor agents, yet their mechanism of action remains largely unknown. Based on the outcome of a virtual screening study, various di- and tri-n-butyltin(IV) carboxylates were synthesized, and their biological properties were evaluated. All synthesized compounds were able to inhibit TrxR selectively within the micromolar range and showed potent antitumor activity against HT-29 and MCF-7 cancer cell lines. Moreover, tin(IV) organometallics were found to strongly induce apoptosis in the BJAB lymphoma cell line. Mass spectrometry and atomic absorption spectroscopy experiments revealed metal binding to proteins, and efficient cellular uptake was observed using a di-n-butyltin(IV) complex as an example.
