RESUMEN
OBJECTIVES: Mycobacterium tuberculosis is a deadly human pathogen that causes the lung disease TB. M. tuberculosis latently infects a third of the world's population, resulting in â¼1.5 million deaths per year. Due to the difficulties and expense of carrying out animal drug trials using M. tuberculosis and rodents, infections of the zebrafish Danio rerio with Mycobacterium marinum have become a useful surrogate. However, the infection methods described to date require specialized equipment and a high level of operator expertise. METHODS: We investigated whether zebrafish larvae could be naturally infected with bioluminescently labelled M. marinum by immersion, and whether infected larvae could be used for rapid screening of anti-mycobacterial compounds using bioluminescence. We used rifampicin and a variety of nitroimidazole-based next-generation and experimental anti-mycobacterial drugs, selected for their wide range of potencies against M. tuberculosis, to validate this model for anti-mycobacterial drug discovery. RESULTS: We observed that five of the six treatments (rifampicin, pretomanid, delamanid, SN30488 and SN30527) significantly reduced the bioluminescent signal from M. marinum within naturally infected zebrafish larvae. Importantly, these same five treatments also retarded the growth of M. tuberculosis in vitro. In contrast, only three of the six treatments tested (rifampicin, delamanid and SN30527) retarded the growth of M. marinum in vitro. CONCLUSIONS: We have demonstrated that zebrafish larvae naturally infected with bioluminescent M. marinum M can be used for the rapid screening of anti-mycobacterial compounds with readily available equipment and limited expertise. The result is an assay that can be carried out by a wide variety of laboratories for minimal cost and without high levels of zebrafish expertise.
Asunto(s)
Antituberculosos/aislamiento & purificación , Antituberculosos/farmacología , Evaluación Preclínica de Medicamentos/métodos , Mycobacterium marinum/efectos de los fármacos , Pez Cebra/microbiología , Animales , Larva/microbiología , Mediciones Luminiscentes , Mycobacterium marinum/crecimiento & desarrollo , Nitroimidazoles/farmacología , Rifampin/farmacología , Coloración y EtiquetadoRESUMEN
Chronic inflammation can lead to the development of cancers and resolution of inflammation is an ongoing challenge. Inflammation can result from dysregulation of the epigenome and a number of compounds that modify the epigenome are in clinical use. In this study the anti-inflammatory and anti-cancer effects of a quinazoline epigenetic-modulator compound were determined in prostate cancer cell lines using a non-hypothesis driven transcriptomics strategy utilising the Affymetrix PrimeView® Human Gene Expression microarray. GATHER and IPA software were used to analyse the data and to provide information on significantly modified biological processes, pathways and networks. A number of genes were differentially expressed in both PC3 and DU145 prostate cancer cell lines. The top canonical pathways that frequently arose across both cell lines at a number of time points included cholesterol biosynthesis and metabolism, and the mevalonate pathway. Targeting of sterol and mevalonate pathways may be a powerful anticancer approach.
Asunto(s)
Colesterol/biosíntesis , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Inhibidores de Histona Desacetilasas/farmacología , Ácido Mevalónico/metabolismo , Neoplasias de la Próstata/genética , Quinazolinas/farmacología , Línea Celular Tumoral , Perfilación de la Expresión Génica , Humanos , Concentración 50 Inhibidora , Masculino , Análisis de Secuencia por Matrices de Oligonucleótidos , Reproducibilidad de los Resultados , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genéticaRESUMEN
The cytolytic protein perforin is a key component of the immune response and is implicated in a number of human pathologies and therapy-induced conditions. A novel series of small molecule inhibitors of perforin function have been developed as potential immunosuppressive agents. The pharmacokinetics and metabolic stability of a series of 16 inhibitors of perforin was evaluated in male CD1 mice following intravenous administration. The compounds were well tolerated 6 h after dosing. After intravenous administration at 5 mg/kg, maximum plasma concentrations ranged from 532 ± 200 to 10,061 ± 12 ng/mL across the series. Plasma concentrations were greater than the concentrations required for in vitro inhibitory activity for 11 of the compounds. Following an initial rapid distribution phase, the elimination half-life values for the series ranged from 0.82 ± 0.25 to 4.38 ± 4.48 h. All compounds in the series were susceptible to oxidative biotransformation. Following incubations with microsomal preparations, a tenfold range in in vitro half-life was observed across the series. The data suggests that oxidative biotransformation was not singularly responsible for clearance of the compounds and no direct relationship between microsomal clearance and plasma clearance was observed. Structural modifications however, do provide some information as to the relative microsomal stability of the compounds, which may be useful for further drug development.
Asunto(s)
Inmunosupresores/farmacocinética , Perforina/antagonistas & inhibidores , Perforina/metabolismo , Animales , Evaluación Preclínica de Medicamentos/métodos , Masculino , Ratones , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismoRESUMEN
OBJECTIVE: We describe how a single intraperitoneal injection of an indoline-derived drug (SN 28127) reduced mouse body weight (25-45% loss) and adipose tissue mass (â¼75%). METHODS AND RESULTS: The reductions in body weight peaked at â¼21-28 days post drug injection and were maintained throughout the study (160 days). The mice ate as much as vehicle-treated control mice. A more potent SN 28127 analog (SN 29220) reversed high-fat diet-induced obesity and type 2 diabetes in C57BL/6J mice on a high-fat diet. Insulin induced a sustained reduction in blood glucose in fasted SN 29220-treated mice compared with the vehicle-treated mice. All drug-treated mice exhibited a transient increase in water intake from â¼10 days post drug injection that lasted for â¼70 days. Following a single injection of (3)H-labeled SN 29220, radioactivity accumulated within 4 h in the liver, bile duct and ileum with little detected in the brain; within 1-2 days, most of the radioactivity was found in the pancreas, spleen, liver, bile duct, stomach, kidneys and white adipose tissue. High levels of glucose were detected in urine collected from SN 29220 but not vehicle-treated C57BL/6J mice at â¼60 days post injection, while fecal triacylglycerols and cholesterol were not different between SN 29220 and vehicle-treated mice. These data lead us to hypothesize that the hepatic system is the primary drug target. Genes involved in fatty acid synthesis (FASn, SCD1 and PPARγ) and appetite stimulation (AGRP) were upregulated at 160 days post drug treatment, indicative of adaptation to reduced body weight. CONCLUSION: We hypothesize that indoline-derived drug-induced chronic toxicity to the hepatic system leads to a reduction in white adipose tissue mass. The mice adapt to this drug-induced toxicity with reduced steady-state body weight. Understanding molecular mechanisms underlying these responses has potential to identify novel targets for prevention and treatment of obesity.
Asunto(s)
Tejido Adiposo/efectos de los fármacos , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Indoles/farmacología , Obesidad/tratamiento farmacológico , Compuestos de Amonio Cuaternario/farmacología , Pérdida de Peso/efectos de los fármacos , Animales , Regulación del Apetito/efectos de los fármacos , Glucemia/metabolismo , Dieta Alta en Grasa , Indoles/síntesis química , Inyecciones Intraperitoneales , Masculino , Ratones , Ratones Endogámicos C57BL , Obesidad/etiología , Radiofármacos/metabolismo , Tritio/metabolismoRESUMEN
BACKGROUND AND PURPOSE: PA-824 is a 2-nitroimidazooxazine prodrug currently in Phase II clinical trial for tuberculosis therapy. It is bioactivated by a deazaflavin (F(420) )-dependent nitroreductase (Ddn) isolated from Mycobacterium tuberculosis to form a des-nitro metabolite. This releases toxic reactive nitrogen species which may be responsible for its anti-mycobacterial activity. There are no published reports of mammalian enzymes bioactivating this prodrug. We have investigated the metabolism of PA-824 following incubation with a subcellular fraction of human liver, in comparison with purified Ddn, M. tuberculosis and Mycobacterium smegmatis. EXPERIMENTAL APPROACH: PA-824 (250 µM) was incubated with the 9000 × g supernatant (S9) of human liver homogenates, purified Ddn, M. tuberculosis and M. smegmatis for metabolite identification by liquid chromatography mass spectrometry analysis. KEY RESULTS: PA-824 was metabolized to seven products by Ddn and M. tuberculosis, with the major metabolite being the des-nitro product. Six of these products, but not the des-nitro metabolite, were also detected in M. smegmatis. In contrast, only four of these metabolites were observed in human liver S9; M3, a reduction product previously proposed as an intermediate in the Ddn-catalyzed des-nitrification and radiolytic reduction of PA-824; two unidentified metabolites, M1 and M4, which were products of M3; and a haem-catalyzed product of imidazole ring hydration (M2). CONCLUSIONS AND IMPLICATIONS: PA-824 was metabolized by des-nitrification in Ddn and M. tuberculosis, but this does not occur in human liver S9 and M. smegmatis. Thus, PA-824 was selectively bioactivated in M. tuberculosis and there was no evidence for 'cross-activation' by human enzymes.
Asunto(s)
Antituberculosos/farmacocinética , Hígado/metabolismo , Mycobacterium smegmatis/efectos de los fármacos , Mycobacterium tuberculosis/efectos de los fármacos , Nitroimidazoles/farmacocinética , Fracciones Subcelulares/metabolismo , Antituberculosos/farmacología , Secuencia de Bases , Biotransformación , Cartilla de ADN , Humanos , Espectrometría de Masas , Mycobacterium smegmatis/metabolismo , Mycobacterium tuberculosis/metabolismo , Nitroimidazoles/farmacologíaRESUMEN
Conditionally replicating adenoviruses (CRAd) 'armed' with prodrug-activating genes have the potential to augment the efficacy of virotherapy. An Escherichia coli nitroreductase (NTR) gene (nfsB) was introduced into the E3B region of the systemically active CRAd ONYX-411, to produce ONYX-411(NTR), which had single agent oncolytic activity equivalent to unarmed virus in vitro and in vivo. A fluorogenic probe (SN 29884) developed to monitor NTR expression revealed robust, durable NTR expression in ONYX-411(NTR) infected neoplastic but not primary human cell lines. NTR expression occurred >24 h post-infection in parallel with fiber and was sensitive to ara-C indicating transcriptional linkage to viral replication. A novel NTR prodrug, the 3,5-dinitrobenzamide-2-bromomustard SN 27686, was shown to be more dose potent and selective than CB 1954 and provided a superior bystander effect in 3D multicellular layer cultures. Its water-soluble phosphate ester SN 28343 was substantially more active than CB 1954 against xenografts containing a minority of stable NTR-expressing cells. A single intravenous dose of ONYX-411(NTR) (10(8) PFU) to nude mice bearing large H1299 xenografts (>350 mm(3)) resulted in tumor-specific NTR expression which increased over time. Despite extensive viral spread by day 14, this conservative virus dose and schedule was unable to control such well-established tumors. However, subsequent administration of SN 28343 resulted in the majority of mice (62.5%) being tumor-free on day 120.
Asunto(s)
Adenoviridae , Antineoplásicos/farmacología , Proteínas de Escherichia coli/biosíntesis , Neoplasias/terapia , Compuestos de Mostaza Nitrogenada/farmacología , Nitrorreductasas/biosíntesis , Viroterapia Oncolítica , Virus Oncolíticos , Profármacos/farmacología , Transducción Genética , Animales , Aziridinas/farmacología , Proteínas de Escherichia coli/genética , Expresión Génica , Humanos , Ratones , Ratones Mutantes , Neoplasias/enzimología , Neoplasias/genética , Nitrorreductasas/genética , Virus Oncolíticos/enzimología , Virus Oncolíticos/genética , Factores de Tiempo , Replicación Viral/efectos de los fármacos , Replicación Viral/genética , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
As part of an ongoing drug development programme, this paper describes the sequence specificity and time course of DNA adduct formation for a series of novel DNA-targeted analogues of cis-diaminedichloroplatinum(II) (cisplatin) (9-aminoacridine-4-carboxamide Pt complexes) in intact HeLa cells. The sequence specificity of DNA damage caused by cisplatin and analogues in human (HeLa) cells was studied using Taq DNA polymerase and a linear amplification/polymerase stop assay. Primer extension is inhibited by a Pt-DNA adduct, and hence the sites of these lesions can be analysed on DNA sequencing gels. The repetitive alphoid DNA sequence was used as the target DNA in human cells. The 9-aminoacridine-4-carboxamide Pt complexes exhibited a markedly different sequence specificity relative to cisplatin and other analogues. The sequence specificity of the 9-aminoacridine-4-carboxamide Pt complexes is shifted away from a preference for runs of guanines. The 9-aminoacridine-4-carboxamide Pt complexes have an enhanced preference for GA dinucleotides. This is the first occasion that an altered DNA sequence specificity has been demonstrated for a cisplatin analogue in human cells. A time course of DNA damage revealed that the DNA-targeted Pt complexes, consisting of four 9-aminoacridine-4-carboxamide Pt complexes and one acridine-4-carboxamide Pt complex, damaged DNA more rapidly compared to cisplatin and non-targeted analogues. A comparison of the time taken to reach half the maximum relative intensity indicated that the DNA-targeted Pt complexes reacted approximately 4-fold faster than cisplatin and the non-targeted analogues.
Asunto(s)
Cisplatino/análogos & derivados , Aductos de ADN/química , Daño del ADN , Sustancias Intercalantes/farmacología , Compuestos Organoplatinos/farmacología , Aminoacridinas/química , Secuencia de Bases , Sitios de Unión , Cisplatino/farmacología , Células HeLa , Humanos , Sustancias Intercalantes/química , Compuestos Organoplatinos/química , Polimerasa Taq , Factores de TiempoRESUMEN
Irreversible inhibitors of the epidermal growth factor receptor (EGFR) are showing promise in clinical trials. This report is the first to show that inhibition of the EGFR tyrosine kinase by an irreversible binder synergizes with cisplatin, at least in EGFR-overexpressing tissue culture cell lines in vitro. Unlike previous synergies demonstrated between ErbB2 blockade and DNA-damaging drugs, the synergy between the irreversible EGFR inhibitor and cisplatin does not appear to involve the repair of DNA-cisplatin adducts. Given the current clinical data, this combination may be of more than theoretical interest.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Cisplatino/administración & dosificación , Aductos de ADN/efectos de los fármacos , Receptores ErbB/antagonistas & inhibidores , Luciferasas/efectos de los fármacos , Morfolinas/administración & dosificación , Animales , Carcinoma de Células Escamosas/enzimología , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , División Celular/efectos de los fármacos , Células Cultivadas , Reparación del ADN/efectos de los fármacos , Esquema de Medicación , Sinergismo Farmacológico , Inhibidores Enzimáticos/administración & dosificación , Fibroblastos/enzimología , Fibroblastos/patología , Humanos , Ratones , Células Tumorales CultivadasRESUMEN
Systemic cytotoxic (anti-proliferative) anticancer drugs rely primarily for their therapeutic effect on cytokinetic differences between cancer and normal cells. One approach aimed at improving the selectivity of tumour cell killing by such compounds is the use of less toxic prodrug forms that can be selectively activated in tumour tissue (tumour-activated prodrugs; TAP). There are several mechanisms potentially exploitable for selective activation. Some utilise unique aspects of tumour physiology such as selective enzyme expression, hypoxia, and low extracellular pH. Others are based on tumour-specific delivery techniques, including activation of prodrugs by exogenous enzymes delivered to tumour cells via monoclonal antibodies (ADEPT), or generated in tumour cells from DNA constructs containing the corresponding gene (GDEPT). Because only a small proportion of the tumour cells may be competent to activate the prodrug, whichever activating mechanism is used, TAP need to be capable of killing activation-incompetent cells as well via a "bystander effect", in order to fully exploit these "activator" cells. A wide variety of chemistries have been explored for the selective activation of TAP. These include reduction of quinones, N-oxides, nitroaromatics and metal complexes by endogenous enzymes or radiation, amide cleavage by endogenous peptidases, and metabolism by a variety of exogenous enzymes, including phosphatases, kinases, amidases and glycosidases.
Asunto(s)
Antineoplásicos/uso terapéutico , Neoplasias/tratamiento farmacológico , Profármacos/efectos de la radiación , Profármacos/uso terapéutico , Animales , Antineoplásicos/efectos de la radiación , Diseño de Fármacos , Enzimas/metabolismo , Epítopos/inmunología , Humanos , Neoplasias/inmunología , Neoplasias/metabolismoRESUMEN
Nitrobenzyl quaternary salts of nitrogen mustards have been previously reported as hypoxia-selective cytotoxins. In this paper we describe the synthesis and evaluation of a series of heterocyclic analogues, including pyrrole, imidazole, thiophene, and pyrazole examples, chosen to cover a range of one-electron reduction potentials (from -277 to -511 mV) and substitution patterns. All quaternary salt compounds were less toxic in vitro than mechlorethamine, and all were more toxic under hypoxic than aerobic conditions, although the differentials were highly variable within the series. The most promising analogue, imidazole 2, demonstrated DNA cross-linking selectively in hypoxic RIF-1 cells, and was active in vivo in combination with radiation or cisplatin. However, 2 also produced unpredictable toxicity in vivo, suggestive of nonspecific nitrogen mustard release, and this has restricted further development of these compounds as hypoxia-selective cytotoxins.
Asunto(s)
Antineoplásicos/síntesis química , Imidazoles/síntesis química , Nitrocompuestos/síntesis química , Compuestos de Mostaza Nitrogenada/síntesis química , Profármacos/síntesis química , Compuestos de Amonio Cuaternario/síntesis química , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Antineoplásicos/toxicidad , Muerte Celular/efectos de los fármacos , Hipoxia de la Célula , Línea Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Ensayo Cometa , Cricetinae , Reactivos de Enlaces Cruzados/síntesis química , Reactivos de Enlaces Cruzados/química , Reactivos de Enlaces Cruzados/farmacología , Reactivos de Enlaces Cruzados/toxicidad , ADN/química , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Imidazoles/química , Imidazoles/farmacología , Imidazoles/toxicidad , Dosis Máxima Tolerada , Ratones , Trasplante de Neoplasias , Nitrocompuestos/química , Nitrocompuestos/farmacología , Nitrocompuestos/toxicidad , Compuestos de Mostaza Nitrogenada/química , Compuestos de Mostaza Nitrogenada/farmacología , Compuestos de Mostaza Nitrogenada/toxicidad , Oxidación-Reducción , Profármacos/química , Profármacos/farmacología , Profármacos/toxicidad , Compuestos de Amonio Cuaternario/química , Compuestos de Amonio Cuaternario/farmacología , Compuestos de Amonio Cuaternario/toxicidad , Células Tumorales CultivadasRESUMEN
QSAR have been developed for the anticancer activity (growth inhibition) of various tumor cells by bis(11-oxo-11H-indeno[1,2-b]quinoline-6-carboxamides), bis(phenazine-1-carboxamides), and bis(naphthalimides). Of the seven QSAR, positive hydrophobic interactions are found in only two examples: bis(naphthalimides) versus human colon cancer cells. This is consistent with other QSAR of anticancer compounds where hydrophobic interactions are found to be unimportant.
Asunto(s)
Antineoplásicos/química , Relación Estructura-Actividad Cuantitativa , 1-Naftilamina/análogos & derivados , 1-Naftilamina/química , Animales , División Celular/efectos de los fármacos , Dimerización , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Imidas/química , Indenos/química , Concentración 50 Inhibidora , Ratones , Fenazinas/química , Quinolinas/química , Quinolonas/química , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
4-Anilinoquinazoline- and 4-anilinopyrido[3,4-d]pyrimidine-6-acrylamides are potent pan-erbB tyrosine kinase inactivators, and one example (CI-1033) is in clinical trial. A series of analogues with a variety of Michael acceptor units at the 6-position were prepared to define the structural requirements for irreversible inhibition. A particular goal was to determine whether additional functions to increase solubility could be appended to the Michael acceptor. Substituted acrylamides were prepared by direct acylation of the corresponding 6-amines with the requisite acid or acid chloride. Vinylsulfonamide derivatives were obtained by acylation of the amines with chloroethylsulfonyl chloride followed by base-promoted elimination. Vinylsulfone and vinylsulfine derivatives were prepared by oxidation and base elimination of a hydroxyethylthio intermediate. The compounds were evaluated for their inhibition of phosphorylation of the isolated EGFR enzyme and for inhibition of EGF-stimulated autophosphorylation of EGFR in A431 cells and of heregulin-stimulated autophosphorylation of erbB2 in MDA-MB 453 cells. Substitution at the nitrogen of the acrylamide was tolerated only with a methyl group; larger substituents were dystherapeutic, and no substitution at all was tolerated at the acrylamide alpha-carbon. In contrast, while electron-donating groups at the acrylamide beta-carbon were not useful, even quite large electron-withdrawing groups (which increase its electrophilicity) were tolerated. A series of derivatives with solubility-enhancing substituents linked to the acrylamide beta-carbon via amides were potent irreversible inhibitors of isolated EGFR (IC50s = 0.4-1.1 nM), with weakly basic morpholine and imidazole derivatives being the best. Vinylsulfonamides were also potent and irreversible inhibitors, but vinylsulfones and vinylsulfines were reversible and only poorly active. Two compounds were evaluated against A431, H125, and MCF-7 xenografts in nude mice but were inferior in these assays to the clinical trial compound CI-1033.
Asunto(s)
Antineoplásicos/síntesis química , Inhibidores Enzimáticos/síntesis química , Receptores ErbB/antagonistas & inhibidores , Pirimidinas/síntesis química , Quinazolinas/síntesis química , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Receptores ErbB/metabolismo , Concentración 50 Inhibidora , Ratones , Fosforilación , Pirimidinas/química , Pirimidinas/farmacología , Quinazolinas/química , Quinazolinas/farmacología , Solubilidad , Relación Estructura-Actividad , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
A series of intercalator-tethered platinum(II) complexes PtLCl(2) have been prepared where L are the diamine ligands N-[2-[(aminoethyl)amino]ethyl]-9-aminoacridine-4-carboxamide, N-[3-[(2-aminoethyl)amino]propyl]-9-aminoacridine-4-carboxamide, N-[4-[(2-aminoethyl)amino]butyl]-9-aminoacridine-4-carboxamide and N-[5-[(aminoethyl)amino]pentyl]-9-aminoacridine-4-carboxamide and N-[6-[(aminoethyl)amino]hexyl]-9-aminoacridine-4-carboxamide. The activity of the complexes was assessed in the CH-1, CH-1cisR, 41M, 41McisR and SKOV-3 cell lines. The compounds with the shorter linker chain lengths are generally the most active against these cell lines and are much more toxic than Pt(en)C1(2). For example, for the n=2 compound the IC(50) values are 0.017 microM (CH-1), 1.7 microM (41M), 1.4 microM (SKOV-3) and the resistance ratios are 51 (CH-1cisR) and 1.6 (41McisR). For the untethered analogue Pt(en)C1(2) the IC(50) values are 2.5 microM (CH-1), 2.9 microM (41M), 45 microM (SKOV-3) and the resistance ratios are 2.8 (CH-1cisR) and 4.1 (41McisR). The very large differential in IC(50) values between the CH-1 and CH-1cisR pair of cell lines for the 9-aminoacridine-4-carboxamide tethered platinum complexes indicates that repair of platinum-induced DNA damage may be a major determinant of the activity of these compounds.
Asunto(s)
Aminoacridinas/farmacología , Antineoplásicos/síntesis química , Compuestos Organoplatinos/farmacología , Aminoacridinas/síntesis química , Aminoacridinas/química , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Supervivencia Celular/efectos de los fármacos , Resistencia a Medicamentos , Femenino , Humanos , Concentración 50 Inhibidora , Espectroscopía de Resonancia Magnética , Ratones , Compuestos Organoplatinos/síntesis química , Compuestos Organoplatinos/química , Células Tumorales CultivadasRESUMEN
A series of 7-oxo-7H-dibenz[f,ij]isoquinoline and 7-oxo-7H-benzo[e]perimidines bearing cationic side chains were prepared from aminoanthraquinones. The perimidines were prepared from 1-aminoanthraquinone by initial condensation with urea or dimethylacetamide. A series of 2-, 4-, 8-, and 11-carboxy derivatives of the dibenzisoquinolines were prepared from aminoanthraquinonecarboxylic acids. The cationic derivatives were prepared from these via amide, amine, or methylene linkers to study the effects of side chain positioning on biological activity. Within the series of carboxamide-linked compounds, the order of increasing cytotoxicity was 8- < 4- < 2- < 11-. The 2- and 4-carboxamides showed substantial growth delays against in vivo subcutaneous colon 38 tumors in mice, but the 11-carboxamide had curative activity in this refractory model and is being investigated further.
Asunto(s)
Antineoplásicos/síntesis química , Supervivencia Celular/efectos de los fármacos , Isoquinolinas/síntesis química , Isoquinolinas/toxicidad , Quinazolinas/síntesis química , Quinazolinas/toxicidad , Animales , Antineoplásicos/química , Antineoplásicos/uso terapéutico , Neoplasias del Colon/tratamiento farmacológico , Doxorrubicina/toxicidad , Diseño de Fármacos , Resistencia a Antineoplásicos , Humanos , Indicadores y Reactivos , Isoquinolinas/química , Isoquinolinas/uso terapéutico , Células Jurkat , Leucemia P388 , Neoplasias Pulmonares , Ratones , Ratones Endogámicos , Modelos Moleculares , Conformación Molecular , Estructura Molecular , Quinazolinas/química , Quinazolinas/uso terapéutico , Relación Estructura-Actividad , Células Tumorales CultivadasRESUMEN
Inhibitors of topoisomerases are widely used in the treatment of cancer, including inhibitors of topoisomerase I (camptothecin analogs such as irinotecan and topotecan) and topoisomerase II (etoposide and doxorubicin). The novel bis-phenazine, XR5944, is a joint inhibitor of topoisomerase I and II as shown by the stabilization of topoisomerase-dependent cleavable complexes. XR5944 demonstrated exceptional activity against human and murine tumor cells in vitro and in vivo. In a range of cell lines XR5944 (IC50 0.04-0.4 nM) was significantly more potent than TAS-103, originally proposed as a joint topoisomerase I and II inhibitor, as well as agents specific for topoisomerase I or II (topotecan, doxorubicin and etoposide). In addition, XR5944 was unaffected by atypical drug resistance and retained significant activity in cells overexpressing P-glycoprotein or multidrug resistance-associated protein. Antitumor efficacy of XR5944 was demonstrated in human carcinoma xenograft models (H69 small cell lung cancer and HT29 colon). In the HT29 model, which is relatively unresponsive to chemotherapy, XR5944 (15 mg/kg i.v., q4dx3) induced tumor regression in the majority of animals (six of eight), whereas TAS-103, dosed at its maximum tolerated dose (45 mg/kg i.v., q7dx3), only induced a delay in tumor growth compared with control animals. In the H69 model, low doses of XR5944 (5 mg/kg i.v., qdx5/week for 2 weeks or 10-15 mg/kg i.v., q4dx3), induced complete tumor regression in the majority of animals. In contrast, topotecan (20 mg/kg i.v., q4dx3) or etoposide (30 mg/kg i.v., q5dx5) only slowed the tumor growth rate. These studies show that XR5944 is a highly active novel anticancer agent that is well tolerated at efficacious doses.
Asunto(s)
Antineoplásicos/farmacología , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Neoplasias del Colon/tratamiento farmacológico , ADN-Topoisomerasas de Tipo II , Isoenzimas/antagonistas & inhibidores , Neoplasias Pulmonares/tratamiento farmacológico , Fenazinas/farmacología , Inhibidores de Topoisomerasa I , Inhibidores de Topoisomerasa II , Aminoquinolinas/metabolismo , Aminoquinolinas/farmacología , Animales , Antígenos de Neoplasias , Antineoplásicos/metabolismo , Antineoplásicos/toxicidad , ADN/química , ADN/metabolismo , ADN-Topoisomerasas de Tipo I/metabolismo , ADN-Topoisomerasas de Tipo II/metabolismo , Proteínas de Unión al ADN , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo , Doxorrubicina/metabolismo , Doxorrubicina/farmacología , Resistencia a Múltiples Medicamentos , Resistencia a Antineoplásicos , Etopósido/metabolismo , Etopósido/farmacología , Femenino , Humanos , Indenos/metabolismo , Indenos/farmacología , Concentración 50 Inhibidora , Inyecciones Intraperitoneales , Inyecciones Intravenosas , Isoenzimas/metabolismo , Ratones , Ratones Desnudos/genética , Ratones Desnudos/metabolismo , Fenazinas/metabolismo , Fenazinas/toxicidad , Inducción de Remisión , Topotecan/metabolismo , Topotecan/farmacología , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The erbB family of receptor tyrosine kinase enzymes, and particularly EGFR and HER2/neu, have become important targets for potential anticancer drugs. The substrate protein binding site theoretically is the more attractive intracellular target on these enzymes, possessing lower homology than the ATP site between different receptor kinases. However, a major breakthrough in this field was the discovery that 4-anilinoquinazolines are potent and selective inhibitors, despite binding at the ATP site. The very tight structure-activity relationships shown by these compounds suggested a clearly-defined binding mode, where the quinazoline ring binds in the adenine pocket and the anilino ring binds in an adjacent, unique lipophilic pocket. A unique cysteine (Cys-773) adjacent to the quinazoline binding site has prompted the development of irreversible inhibitors that target this residue. Three 4-anilinoquinazoline analogues (two reversible and one irreversible inhibitor) have been evaluated clinically as anticancer drugs. Data from the most advanced, the reversible inhibitor Iressa, suggest that this class of compounds may be of value in cancer chemotherapy.
Asunto(s)
Inhibidores Enzimáticos/farmacología , Receptores ErbB/antagonistas & inhibidores , Quinazolinas/farmacología , Receptor ErbB-2/antagonistas & inhibidores , Adenosina Trifosfato/metabolismo , Simulación por Computador , Relación Estructura-ActividadRESUMEN
Recent work on a number of different classes of anticancer agents that alkylate DNA in the minor groove is reviewed. There has been much work with nitrogen mustards, where attachment of the mustard unit to carrier molecules can change the normal patterns of both regio- and sequence-selectivity, from reaction primarily at most guanine N7 sites in the major groove to a few adenine N3 sites at the 3'-end of poly(A/T) sequences in the minor groove. Carrier molecules discussed for mustards are intercalators, polypyrroles, polyimidazoles, bis(benzimidazoles), polybenzamides and anilinoquinolinium salts. In contrast, similar targeting of pyrrolizidine alkylators by a variety of carriers has little effect of their patterns of alkylation (at the 2-amino group of guanine). Recent work on the pyrrolobenzodiazepine and cyclopropaindolone classes of natural product minor groove binders is also reviewed.
Asunto(s)
Alquilantes/química , Alquilantes/metabolismo , ADN/metabolismo , Netropsina/análogos & derivados , Alquilantes/farmacología , Animales , Antramicina/química , Antramicina/metabolismo , Antramicina/farmacología , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/metabolismo , Antibióticos Antineoplásicos/farmacología , Benzofuranos/química , Benzofuranos/metabolismo , Benzofuranos/farmacología , Bisbenzimidazol/química , Bisbenzimidazol/metabolismo , Clorambucilo/química , Clorambucilo/metabolismo , Ácidos Ciclohexanocarboxílicos/química , Ácidos Ciclohexanocarboxílicos/metabolismo , Ácidos Ciclohexanocarboxílicos/farmacología , Ciclohexenos , ADN/química , ADN/efectos de los fármacos , Distamicinas/química , Distamicinas/metabolismo , Duocarmicinas , Humanos , Indoles/química , Indoles/metabolismo , Indoles/farmacología , Netropsina/química , Netropsina/metabolismo , Compuestos de Mostaza Nitrogenada/química , Compuestos de Mostaza Nitrogenada/metabolismo , Compuestos de Mostaza Nitrogenada/farmacología , Pirroles/química , Pirroles/metabolismo , Relación Estructura-ActividadRESUMEN
Bis(9-methylphenazine-1-carboxamides) joined by a variety of dicationic (CH(2))(n)()NR(CH(2))(m)NR(CH(2))(n) linkers of varying length (carboxamide N-N distances from 11.0 to 18.4 A) and rigidity were prepared by reaction of 9-methylphenazine-1-carboxylic acid imidazolide with the appropriate polyamines. The compounds were evaluated for growth inhibitory properties in P388 leukemia, Lewis lung carcinoma, and wild-type (JL(C)) and mutant (JL(A) and JL(D)) forms of human Jurkat leukemia with low levels of topoisomerase II (topo II). The compounds all had IC(50) ratios of <1 in the resistant Jurkat lines, consistent with topo II inhibition not being the primary mechanism of action. Analogues joined by an (CH(2))(2)NR(CH(2))(2)NR(CH(2))(2) linker were extremely potent cytotoxins, with selectivity toward the human cell lines, but absolute potencies declined sharply from R = H through R = Me to R = Pr and Bu. In contrast, (CH(2))(2)NR(CH(2))(3)NR(CH(2))(2) compounds showed reverse effects, with the R = Me analogue being more potent than the R = H one as well as being the most potent in the series [IC(50) in JL(C) cells 0.08 nM; superior to that for the clinical bis(naphthalimide) LU 79553]. Overall, the IC(50)s of analogues with linker chains (CH(2))(n)NH(CH(2))(m)NH(CH(2))(n) were inversely proportional to linker length. Constraining the rigidity of the linker chain by incorporating a piperazine ring did not decrease potency significantly. A representative compound bound tightly to DNA with high selectivity for GC sites, compatible with recent work suggesting that compounds of this type place their side chains in the major groove, making specific contacts with guanine bases. Representative compounds were susceptible to transport mediated resistance, being much less effective in cells that overexpressed P-glycoprotein. Overall the results suggest these compounds have a similar mode of action, mediated primarily by poisoning of topo I (possibly with some involvement of topo II). The bis(9-methylphenazine-1-carboxamides) show very high in vitro growth inhibitory potencies compared to their monomeric analogues. Two compounds showed in vivo activity in murine colon 38 syngeneic and HT29 human colon tumor xenograft models using intraperitoneal dosing.
Asunto(s)
Amidas/síntesis química , Antineoplásicos/síntesis química , Inhibidores Enzimáticos/síntesis química , Fenazinas/síntesis química , Inhibidores de Topoisomerasa I , Inhibidores de Topoisomerasa II , Amidas/química , Amidas/farmacología , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Cationes Bivalentes , ADN-Topoisomerasas de Tipo I/química , ADN-Topoisomerasas de Tipo II/química , ADN Superhelicoidal/química , Ensayos de Selección de Medicamentos Antitumorales , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Humanos , Concentración 50 Inhibidora , Ratones , Ratones Endogámicos C57BL , Ratones Desnudos , Fenazinas/química , Fenazinas/farmacología , Relación Estructura-Actividad , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
The tricyclic carboxamide N-[2-(dimethylamino)ethyl]acridine-4-carboxamide (DACA) is a DNA-intercalating agent capable of inhibiting both topoisomerases I and II and is currently in Phase II clinical trial. Many related analogues have been developed, but despite their potent in vitro cytotoxicities, they exhibit poor extravascular distribution. As part of an ongoing drug development program to obtain related "minimal intercalators" with lower DNA association constants, we have compared the biodistribution and metabolite profiles of the prototype compound, DACA, with three analogues to aid rational drug selection. All of these compounds share a common structural feature, N-dimethyl side chain, which was radiolabeled with the positron-emitting radioisotope, carbon-11. This strategy was selected because it allows promising candidates emerging from preclinical studies in animals to be evaluated rapidly in humans using positron emission tomography (PET). The acridine DACA, the phenazine SN 23490, the pyridoquinoline SN 23719, and the dibenzodioxin SN 23935 were found to be cytotoxic in in vitro assays with an IC50 of 1.4-1.8 microM, 0.4-0.6 microM, 1.3-1.6 microM, and 24-36 microM, respectively, in HT29, U87MG, and A375M cell lines. Ex vivo biodistribution studies with carbon-11 radiolabeled compounds in mice bearing human tumor xenografts showed rapid clearance of 11C-radioactivity (parent drug and metabolites) from blood and the major organs. Rapid hepatobiliary clearance and renal excretion were also observed. There was low [<5% of injected dose/gram (%ID/g)] and variable uptake of 11C-radioactivity in three tumor types for all of the compounds. Tumor (U87MG) to blood 11C-radioactivity for [11C]DACA, [11C](9-methoxyphenazine-1-carboxamide (SN 23490), [11C]2-(4-pyridyl)quinoline-8-carboxamide (SN 23719), and [11C]dibenzo[1,4]dioxin-1-carboxamide (SN 23935) at 30 min were 2.9 +/- 1.1, 2.3 +/- 0.6, 2.6 +/- 0.6, and 0.7 +/- 0.2, respectively. For SN 23719, the distribution of 11C-radioactivity in normal tissues and tumors determined ex vivo was in broad agreement with that determined in vivo by whole body PET scanning. [11C]DACA was rapidly and extensively metabolized to several plasma metabolites and a major tumor metabolite. In contrast, [11C]SN 23935, [11C]SN 23490, and [11C]SN 23719 showed less extensive metabolism. In the tumor samples, the parent [11C]DACA and [11C]SN 23935 represented between 0.3 and 1.5%ID/g, whereas [11C]SN 23490 and [11C]SN 23719 represented between 1.5 and 2.8%ID/g. In conclusion, by using a strategy with 11C-labeling, we have determined the tissue distribution and metabolic stability of novel tricyclic carboxamides with the view of selecting analogues with potentially better in vivo activity against solid tumors. SN 23490 and SN 23719 had more favorable distribution and metabolic stability compared with DACA and SN 23935 and may warrant further development. The radiolabeling strategy used allows ex vivo and in vivo evaluation of promising anticancer agents in animals and offers the potential of rapid translation to studies in humans using PET.
Asunto(s)
Acridinas/farmacocinética , Antineoplásicos/farmacocinética , Sustancias Intercalantes/farmacocinética , Acridinas/química , Acridinas/metabolismo , Acridinas/toxicidad , Animales , Antineoplásicos/metabolismo , Antineoplásicos/toxicidad , Radioisótopos de Carbono , Femenino , Glioma/tratamiento farmacológico , Glioma/metabolismo , Células HT29 , Humanos , Concentración 50 Inhibidora , Sustancias Intercalantes/metabolismo , Sustancias Intercalantes/toxicidad , Marcaje Isotópico , Melanoma/tratamiento farmacológico , Melanoma/metabolismo , Ratones , Ratones Endogámicos ICR , Ratones Desnudos , Distribución Tisular , Tomografía Computarizada de Emisión , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Two gametocyte-producing isolates of Plasmodium falciparum, KT1 arid KT3, were cultivated in vitro. On day 11 of cultivation, pure gametocytes containing stage II, III and IV were used to test the gametocytocidal activity of 9-anilinoacridines that had previously demonstrated their activity against the asexual stage of the parasite. After drug exposure for 48 h, gametocytes were maintained without drugs for another 2 days before thin films were prepared for parasite counting. Gametocytocidal activities of 13 analogs of 9-anilinoacridine were observed with 50% inhibitory concentrations in the range of 0.6 microM to> 100 microM The most active compound was 1'-CH2NMe2-9-anilinoacridine. Anilinoacridine derivatives with 3,6-diamino substitution had reduced gametocytocidal activity in contrast to their enhancing effect against the asexual forms. Morphological abnormalities of gametocytes were observed following drug exposure.