RESUMEN
In terms of drug disposal and metabolism SDR21C1 (carbonyl reductase 1; CBR1) exerts an assorted substrate spectrum among a large variety of clinically relevant substances. Additionally, this short-chain dehydrogenase/reductase is extensively expressed in most tissues of the human body, thus underpinning its role in xenobiotic metabolism. Reduction of the chemotherapeutic daunorubicin (DAUN) to daunorubicinol (DAUNol) is a prominent example of its metabolic properties in terms of chemoresistance and cardiotoxicity. The hop-derived prenylated chalcone xanthohumol (XN) and its physiological metabolites isoxanthohumol (IX) and 8-prenylnaringenin (8-PN) have previously been reported to inhibit other DAUN reducing reductases and dehydrogenases including AKR1B1 and AKR1B10. Also with regard to their effects by means of interacting with cancer-related molecular pathways, XN and related prenylated flavonoids in particular have been in the focus of recent studies. In this study, inhibitory properties of these substances were examined with CBR1-mediated 2,3-hexanedione and DAUN reduction. All substances tested in this study turned out to efficiently inhibit recombinant human CBR1 within a low micromolar to submicromolar range. Among the substances tested, 8-PN turned out to be the most effective inhibitor when using 2,3-hexanedione as a substrate (Ki(app)â¯=â¯180⯱â¯20â¯nM). Inhibition rates of recombinant CBR1-mediated DAUN reduction were somewhat weaker with IC50-values ranging from 11 to 20⯵M. XN, IX and 8-PN also efficiently inhibited DAUN reduction by SW480 colon adenocarcinoma cytosol (IC50â¯=â¯3.71⯱â¯0.26⯵M with 8-PN as inhibitor). This study identifies prenylated inhibitors, which might potentially interact with endogenous CBR1-driven (de-)toxication systems.
Asunto(s)
Oxidorreductasas de Alcohol/metabolismo , Flavanonas/química , Flavonoides/química , Propiofenonas/química , Xantonas/química , Oxidorreductasas de Alcohol/antagonistas & inhibidores , Oxidorreductasas de Alcohol/genética , Línea Celular Tumoral , Chalconas/química , Daunorrubicina/química , Daunorrubicina/metabolismo , Flavanonas/metabolismo , Flavonoides/metabolismo , Hexanonas/química , Hexanonas/metabolismo , Humanos , Concentración 50 Inhibidora , Cinética , Oxidación-Reducción , Propiofenonas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidad por Sustrato , Xantonas/metabolismoRESUMEN
Hop-derived compounds have been subjected to numerous biomedical studies investigating their impact on a wide range of pathologies. Isomerised bitter acids (isoadhumulone, isocohumulone and isohumulone) from hops, used in the brewing process of beer, are known to inhibit members of the aldo-keto-reductase superfamily. Aldo-keto-reductase 1B10 (AKR1B10) is upregulated in various types of cancer and has been reported to promote carcinogenesis. Inhibition of AKR1B10 appears to be an attractive means to specifically treat RAS-dependent malignancies. However, the closely related reductases AKR1A1 and AKR1B1, which fulfil important roles in the detoxification of endogenous and xenobiotic carbonyl compounds oftentimes crossreact with inhibitors designed to target AKR1B10. Accordingly, there is an ongoing search for selective AKR1B10 inhibitors that do not interact with endogeneous AKR1A1 and AKR1B1-driven detoxification systems. In this study, unisomerised α-acids (adhumulone, cohumulone and n-humulone) were separated and tested for their inhibitory potential on AKR1A1, AKR1B1 and AKR1B10. Also AKR1B10-mediated farnesal reduction was effectively inhibited by α-acid congeners with Ki-values ranging from 16.79 ± 1.33 µM (adhumulone) to 3.94 ± 0.33 µM (n-humulone). Overall, α-acids showed a strong inhibition with selectivity (115â»137 fold) for AKR1B10. The results presented herein characterise hop-derived α-acids as a promising basis for the development of novel and selective AKR1B10-inhibitors.
Asunto(s)
Aldehído Reductasa/antagonistas & inhibidores , Ciclohexanonas/farmacología , Ciclohexenos/farmacología , Terpenos/farmacología , Aldehído Reductasa/metabolismo , Aldo-Ceto Reductasas , Evaluación Preclínica de Medicamentos , Ensayos de Selección de Medicamentos Antitumorales , Farnesol/análogos & derivados , Farnesol/química , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Humanos , Humulus/químicaRESUMEN
Xanthohumol (XN), a prenylated chalcone unique to hops (Humulus lupulus) and two derived prenylflavanones, isoxanthohumol (IX) and 8-prenylnaringenin (8-PN) gained increasing attention as potential anti-diabetic and cancer preventive compounds. Two enzymes of the aldo-keto reductase (AKR) superfamily are notable pharmacological targets in cancer therapy (AKR1B10) and in the treatment of diabetic complications (AKR1B1). Our results show that XN, IX and 8-PN are potent uncompetitive, tight-binding inhibitors of human aldose reductase AKR1B1 (Ki = 15.08 µM, 0.34 µM, 0.71 µM) and of human AKR1B10 (Ki = 20.11 µM, 2.25 µM, 1.95 µM). The activity of the related enzyme AKR1A1 was left unaffected by all three compounds. This is the first time these three substances have been tested on AKRs. The results of this study may provide a basis for further quantitative structure?activity relationship models and promising scaffolds for future anti-diabetic or carcinopreventive drugs.
Asunto(s)
Aldo-Ceto Reductasas/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Flavanonas/farmacología , Flavonoides/farmacología , Humulus/química , Propiofenonas/farmacología , Xantonas/farmacología , Aldo-Ceto Reductasas/metabolismo , Sitios de Unión/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/química , Flavanonas/química , Flavonoides/química , Humanos , Estructura Molecular , Propiofenonas/química , Relación Estructura-Actividad , Xantonas/químicaRESUMEN
The clinical application of anthracyclines, like daunorubicin and doxorubicin, is limited by two factors: dose-related cardiotoxicity and drug resistance. Both have been linked to reductive metabolism of the parent drug to their metabolites daunorubicinol and doxorubicinol, respectively. These metabolites show significantly less anti-neoplastic properties as their parent drugs and accumulate in cardiac tissue leading to chronic cardiotoxicity. Therefore, we aimed to identify novel and potent natural inhibitors for anthracycline reductases, which enhance the anticancer effect of anthracyclines by preventing the development of anthracycline resistance. Human enzymes responsible for the reductive metabolism of daunorubicin were tested for their sensitivity towards anthrachinones, in particular emodin and anthraflavic acid. Intense inhibition kinetic data for the most effective daunorubicin reductases, including IC50- and Ki-values, the mode of inhibition, as well as molecular docking, were compiled. Subsequently, a cytotoxicity profile and the ability of emodin to reverse daunorubicin resistance were determined using multiresistant A549 lung cancer and HepG2 liver cancer cells. Emodin potently inhibited the four main human daunorubicin reductases in vitro. Further, we could demonstrate that emodin is able to synergistically sensitize human cancer cells towards daunorubicin at clinically relevant concentrations. Therefore, emodin may yield the potential to enhance the therapeutic effectiveness of anthracyclines by preventing anthracycline resistance via inhibition of the anthracycline reductases. In symphony with its known pharmacological properties, emodin might be a compound of particular interest in the management of anthracycline chemotherapy efficacy and their adverse effects.
Asunto(s)
Antibióticos Antineoplásicos/farmacología , Daunorrubicina/farmacología , Resistencia a Antineoplásicos/efectos de los fármacos , Emodina/farmacología , Oxidorreductasas/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/farmacología , Antraquinonas/farmacología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Sinergismo Farmacológico , Humanos , Simulación del Acoplamiento Molecular , Oxidorreductasas/metabolismoRESUMEN
Curcumin is a major component of the plant Curcuma longa L. It is traditionally used as a spice and coloring in foods and is an important ingredient in curry. Curcuminoids have anti-oxidant and anti-inflammatory properties and gained increasing attention as potential neuroprotective and cancer preventive compounds. In the present study, we report that curcumin is a potent tight-binding inhibitor of human carbonyl reductase 1 (CBR1, Ki=223 nM). Curcumin acts as a non-competitive inhibitor with respect to the substrate 2,3-hexandione as revealed by plotting IC50-values against various substrate concentrations and most likely as a competitive inhibitor with respect to NADPH. Molecular modeling supports the finding that curcumin occupies the cofactor binding site of CBR1. Interestingly, CBR1 is one of the most effective human reductases in converting the anthracycline anti-tumor drug daunorubicin to daunorubicinol. The secondary alcohol metabolite daunorubicinol has significantly reduced anti-tumor activity and shows increased cardiotoxicity, thereby limiting the clinical use of daunorubicin. Thus, inhibition of CBR1 may increase the efficacy of daunorubicin in cancer tissue and simultaneously decrease its cardiotoxicity. Western-blots demonstrated basal expression of CBR1 in several cell lines. Significantly less daunorubicin reduction was detected after incubating A549 cell lysates with increasing concentrations of curcumin (up to 60% less with 50 µM curcumin), suggesting a beneficial effect in the co-treatment of anthracycline anti-tumor drugs together with curcumin.
Asunto(s)
Aldehído Reductasa/metabolismo , Curcumina/metabolismo , Oxidorreductasas de Alcohol/metabolismo , Aldo-Ceto Reductasas , Antineoplásicos/metabolismo , Antineoplásicos/farmacología , Sitios de Unión , Línea Celular Tumoral , Daunorrubicina/análogos & derivados , Daunorrubicina/metabolismo , Humanos , NADP/metabolismo , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismoRESUMEN
The purpose of this study was to investigate the origin and function of the aldo-keto reductase (AKR) superfamily as enzymes involved in the detoxification of xenobiotics. We used the cyanobacterium Synechocystis sp. PCC 6803 as a model organism and sequence alignments to find bacterial AKRs with highest identity to human enzymes. Disappearance of NADPH was monitored spectrophotometrically to calculate enzymatic activity. The molecular weight of the native protein was determined by size exclusion chromatography. Substrate docking was performed by SwissDock. Sequence alignments identified the NADPH-dependent AKR3G1 having 41.5 and 40% identity with the human enzymes AKR1B1 and AKR1B10, respectively. Highest enzymatic efficiency was observed with 4-oxonon-2-enal (4-ONE; k(cat)/K(m), 561 s(-1) mM(-1)) and 4-hydroxynonenal (k(cat)/K(m), 26.5 s(-1) mM(-1)), respectively. P74308 is the most efficient enzyme for 4-ONE discovered until now. Cooperativity of this monomeric enzyme was observed with some substrates. Enzyme inactivation or oligomerization as possible explanations for nonhyperbolic enzyme kinetics were ruled out by Selwyn's test and gel filtration. The role of the little investigated carbonyl-reducing enzymes in detoxification seems to be in fact a very old process with rarely observed nonhyperbolic enzyme kinetics as an adaptation mechanism to higher concentrations of reactive oxygen species.
Asunto(s)
Aldehído Reductasa/metabolismo , Proteínas Bacterianas/metabolismo , Synechocystis/enzimología , Aldehído Reductasa/química , Aldehído Reductasa/genética , Aldehídos/metabolismo , Aldo-Ceto Reductasas , Secuencia de Aminoácidos , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Dominio Catalítico , Productos Finales de Glicación Avanzada/metabolismo , Humanos , Cinética , Ligandos , Peroxidación de Lípido , Modelos Moleculares , Datos de Secuencia Molecular , Conformación Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homología de Secuencia de Aminoácido , Synechocystis/genéticaRESUMEN
The microsomal enzyme 11ß-hydroxysteroid deydrogenase type 1 (11ß-HSD1) catalyzes the interconversion of glucocorticoid receptor-inert cortisone to receptor- active cortisol, thereby acting as an intracellular switch for regulating the access of glucocorticoid hormones to the glucocorticoid receptor. There is strong evidence for an important aetiological role of 11ß-HSD1 in various metabolic disorders including insulin resistance, diabetes type 2, hypertension, dyslipidemia and obesity. Hence, modulation of 11ß-HSD1 activity with selective inhibitors is being pursued as a new therapeutic approach for the treatment of the metabolic syndrome. Since tea has been associated with health benefits for thousands of years, we sought to elucidate the active principle in tea with regard to diabetes type 2 prevention. Several teas and tea specific polyphenolic compounds were tested for their possible inhibition of cortisone reduction with human liver microsomes and purified human 11ß-HSD1. Indeed we found that tea extracts inhibited 11ß-HSD1 mediated cortisone reduction, where green tea exhibited the highest inhibitory potency with an IC50 value of 3.749 mg dried tea leaves per ml. Consequently, major polyphenolic compounds from green tea, in particular catechins were tested with the same systems. (-)-Epigallocatechin gallate (EGCG) revealed the highest inhibition of 11ß-HSD1 activity (reduction: IC50â=â57.99 µM; oxidation: IC50â=â131.2 µM). Detailed kinetic studies indicate a direct competition mode of EGCG, with substrate and/or cofactor binding. Inhibition constants of EGCG on cortisone reduction were Kiâ=â22.68 µM for microsomes and Kiâ=â18.74 µM for purified 11ß-HSD1. In silicio docking studies support the view that EGCG binds directly to the active site of 11ß-HSD1 by forming a hydrogen bond with Lys187 of the catalytic triade. Our study is the first to provide evidence that the health benefits of green tea and its polyphenolic compounds may be attributed to an inhibition of the cortisol producing enzyme 11ß-HSD1.