RESUMEN
Animal experiments remain essential to understand the fundamental mechanisms underpinning malignancy and to discover improved methods to prevent, diagnose and treat cancer. Excellent standards of animal care are fully consistent with the conduct of high quality cancer research. Here we provide updated guidelines on the welfare and use of animals in cancer research. All experiments should incorporate the 3Rs: replacement, reduction and refinement. Focusing on animal welfare, we present recommendations on all aspects of cancer research, including: study design, statistics and pilot studies; choice of tumour models (e.g., genetically engineered, orthotopic and metastatic); therapy (including drugs and radiation); imaging (covering techniques, anaesthesia and restraint); humane endpoints (including tumour burden and site); and publication of best practice.
Asunto(s)
Experimentación Animal/normas , Bienestar del Animal/normas , Neoplasias/patología , Neoplasias/terapia , Guías de Práctica Clínica como Asunto , Algoritmos , Experimentación Animal/ética , Bienestar del Animal/ética , Bienestar del Animal/organización & administración , Animales , Biomarcadores Farmacológicos/análisis , Investigación Biomédica/ética , Investigación Biomédica/legislación & jurisprudencia , Investigación Biomédica/organización & administración , Investigación Biomédica/normas , Línea Celular Transformada , Diagnóstico por Imagen , Modelos Animales de Enfermedad , Femenino , Humanos , Masculino , Ratones , Trasplante de Neoplasias/métodos , Trasplante de Neoplasias/patología , Trasplante de Neoplasias/normas , Neoplasias/diagnóstico , Neoplasias/genética , Resultado del Tratamiento , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The development of agents that target tumour vasculature is ultimately dependent on the availability of appropriate preclinical screening assays. Several quantitative angiogenesis assays exist, each with its own unique characteristics and disadvantages. In this review we discuss some of the commonly used assays, their methodological pitfalls and current use. The corneal micropocket and the CAM assay are well established. However, the matrix-implant assays have the potential advantage of replicating the hypoxic tumour microenvironment, thus making them suitable for the study of tumour angiogenesis. The ideal quantitative angiogenesis assay does not exist and the use of two complimentary quantitative assays, such as a matrix implant assay and a microcirculatory preparation like the CAM or corneal micropocket assay, provides the best compromise. Newer models like the hollow-fibre assay are being developed and older ones refined. Assay systems should reflect distinct disease processes. Thus it is appropriate to develop assays that study exclusively pro- or anti-angiogenic compounds or anti-vascular agents. Criticisms of currently available screening systems are that the predictive value of current screening systems remains to be established as anti-angiogenic agents are still in clinical development. Anti-angiogenic agents are likely to be most effective as chronic therapy for remission maintenance in the metastatic setting or as adjuvant therapy in patients at high risk of relapse, an important clinical aspect not addressed in animal models of tumour angiogenesis. Histological analysis still provides the most detailed information on in vivo angiogenesis. However, angiogenesis is a dynamic process and assays that permit continuous monitoring of the angiogenic response and provide information on the physiological characteristics of new vessels will be distinctly advantageous over older systems. The development of non-invasive techniques for quantitation of angiogenesis will greatly facilitate this process.
Asunto(s)
Neovascularización Patológica/diagnóstico , Animales , Técnicas Citológicas , Diagnóstico por Imagen , Modelos Animales de Enfermedad , Humanos , Métodos , Neovascularización Patológica/etiologíaRESUMEN
BACKGROUND: A new series of imidazothioxanthones has recently been synthesized as potential anticancer agents with the aim of overcoming drug resistance. The route of synthesis and DNA-binding properties of the compounds were reported previously. This paper describes the general structure-activity relationships for the class of imidazothioxanthones in panels of human and murine tumor cell lines in vitro, and the in vivo activity against human and murine solid tumors of the most potent compound, N-[3-(Dimethylamino)propylo]-11-oxo-11H-benzothiopyrano [3',2':2,3]pyrido[1,2-a]imidazo-2-carboxamide (10a). In addition, the interaction between compound 10a and DNA is also considered in terms of molecular mechanics methods and flexible docking techniques. MATERIALS AND METHODS: The cytotoxicity of compounds 10a, 11-oxo-N-[2-(pyrrolidino)ethylo]-11H-benzothiopyrano [3',2':2,3]pyrido[1,2-a]imidazo-2-carboxamide, 11-oxo-N-[2-(piperidino)ethylo]-11H-benzothiopyrano [3',2':2,3]pyrido[1,2-a]imidazo-2-carboxamide and N-[2-(morpholino)ethylo]-11-oxo-11H-benzothiopyrano [3',2':2,3]pyrido[1,2-a]imidazo-2-carboxamide (10c-10e) was assessed in human tumor cell lines and xenografts using the sulforhodamine B assay, MTT assay and the clonogenic assay. The human ovarian xenograft, PXN/109TC, two human breast carcinomas, MT-1 and MCF-7, and the murine colon adenocarcinoma, MAC15A were used for the in vivo testing of compound 10a. In addition, the interaction between compound 10a and DNA is also considered in terms of molecular mechanics methods and flexible docking techniques. RESULTS: Two compounds, 10a and 10c, showed cytotoxic activity below 10 mM in the NCI in vitro screen of 60 human tumor cell lines. The IC50 value of compound 10a was 6.8 mM and that of 10c, 8.3 mM. In addition, both compounds possessed differential activity against leukemia, colon and mammary cancer. The activity pattern was confirmed in two further screens using monolayer and clonogenic, assays. In vivo antitumor studies showed that 10a was active against the human mammary carcinoma MT-1 and murine colon cancer MAC15A. Marginal activity was observed in human ovarian cancer model PXN/109T/C and the compound was inactive in human mammary cancer MCF-7. CONCLUSION: The results warrant further in vivo testing of 10a in additional human solid tumor models. The molecular modeling showed that the planarity of the chromophore and the side-chain conformation could assist the insertion of compound 10a between the base pairs of the double helix. On the other hand, docking to the nucleotide sequence GGAATTGCCTCA suggested that the molecule could also act as a minor groove binder.
Asunto(s)
Antineoplásicos/farmacología , Imidazoles/farmacología , Xantonas/farmacología , Adenocarcinoma/tratamiento farmacológico , Animales , Antineoplásicos/química , Neoplasias de la Mama/tratamiento farmacológico , Línea Celular Tumoral , Neoplasias del Colon/tratamiento farmacológico , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Humanos , Imidazoles/química , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Modelos Moleculares , Neoplasias/tratamiento farmacológico , Neoplasias Ováricas/tratamiento farmacológico , Xantonas/química , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
2-(4-Aminophenyl)benzothiazoles represent a potent and highly selective class of antitumour agent. In vitro, sensitive carcinoma cells deplete 2-(4-aminophenyl)benzothiazoles from nutrient media; cytochrome P450 1A1 activity, critical for execution of antitumour activity, and protein expression are powerfully induced. 2-(4-Amino-3-methylphenyl)benzothiazole-derived covalent binding to cytochrome P450 1A1 is reduced by glutathione, suggesting 1A1-dependent production of a reactive electrophilic species. In vitro, 2-(4-aminophenyl)benzothiazole-generated DNA adducts form in sensitive tumour cells only. At concentrations >100 nM, adducts were detected in DNA of MCF-7 cells treated with 2-(4-amino-3-methylphenyl)-5-fluorobenzothiazole (5F 203). 5F 203 (1 microM) led to the formation of one major and a number of minor adducts. However, treatment of cells with 10 microM 5F 203 resulted in the emergence of a new dominant adduct. Adducts accumulated steadily within DNA of MCF-7 cells exposed to 1 microM 5F 203 between 2 and 24 h. Concentrations of the lysylamide prodrug of 5F 203 (Phortress) > or = 100 nM generated adducts in the DNA of sensitive MCF-7 and IGROV-1 ovarian cells. At 1 microM, one major Phortress-derived DNA adduct was detected in these two sensitive phenotypes; 10 microM Phortress led to the emergence of an additional major adduct detected in the DNA of MCF-7 cells. Inherently resistant MDA-MB-435 breast carcinoma cells incurred no DNA damage upon exposure to Phortress (< or = 10 microM, 24 h). In vivo, DNA adducts accumulated within sensitive ovarian IGROV-1 and breast MCF-7 xenografts 24 h after treatment of mice with Phortress (20 mg kg(-1)). Moreover, Phortress-derived DNA adduct generation distinguished sensitive MCF-7 tumours from inherently resistant MDA-MB-435 xenografts implanted in opposite flanks of the same mouse.
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Antineoplásicos/farmacología , Aductos de ADN/efectos de los fármacos , Tiazoles/farmacología , Animales , Antineoplásicos/uso terapéutico , Benzotiazoles , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Neoplasias Experimentales/tratamiento farmacológico , Tiazoles/uso terapéutico , Factores de Tiempo , Células Tumorales CultivadasRESUMEN
BACKGROUND: Vinflunine is a novel Vinca alkaloid currently undergoing Phase II clinical trials, which have previously demonstrated anti-vascular effects in a transplantable murine colon adenocarcinoma model. Previous studies with compounds showing similar effects in combination with standard anti-cancer agents have demonstrated an improved efficacy relative to the standard agents. MATERIALS AND METHODS: In this study the synergistic effects of administering vinflunine in combination with either Cisplatin (CPL) or 5-fluorouracil (5-FU) were investigated in a well-differentiated transplantable murine colon adenocarcinoma model (MAC 29). RESULTS: Vinflunine significantly potentiated the anti-tumour effects of CPL, but had little effect in combination with 5-FU. Using Hoescht 33342 dye labelling of the functional vasculature, clear evidence of vascular shutdown was seen for treatment groups including vinflunine. CONCLUSION: These data demonstrate that the combination of vinflunine and CPL has significant preclinical anti-tumour activity against a transplantable murine adenocarcinoma model that is related to the anti-vascular effects of vinflunine.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Cisplatino/uso terapéutico , Fluorouracilo/uso terapéutico , Vinblastina/análogos & derivados , Vinblastina/uso terapéutico , Adenocarcinoma/patología , Animales , Cisplatino/administración & dosificación , Neoplasias del Colon/tratamiento farmacológico , Neoplasias del Colon/patología , Modelos Animales de Enfermedad , Femenino , Fluorouracilo/administración & dosificación , Ratones , Ratones Endogámicos , Trasplante de Neoplasias , Células Tumorales Cultivadas , Vinblastina/administración & dosificaciónRESUMEN
Pharmacogenetic analysis of polymorphisms in drug metabolizing enzymes is currently generating considerable interest as a means of individualizing patient therapy. Recent studies have suggested that patients that are homozygous for a polymorphic variant (a C to T transition at position 609 of the cDNA sequence) of the enzyme NAD(P)H:quinone oxidoreductase (NQO1) may be resistant to Mitomycin C (MMC). Genotyping of a panel of 54 human tumor xenografts by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), classified tumors as wild type (40/54), heterozygotes (11/54), and homozygous mutants (3/54). Previously, 37 of these tumors had been characterized in terms of their response to MMC in vivo, and in this study, a further nine tumor xenografts have been characterized in terms of their response to MMC. No correlation could be found between the NQO1 polymorphic status of xenografts and their response to MMC in vivo. In terms of genotype/phenotype relationships, NQO1 activity in tumors genotyped as wild type, heterozygotes, and homozygous mutants were 311.1 +/- 421.9 (N = 40), 76.9 +/- 109.5 (N = 11), and 0.2 +/- 0.17 (N = 3) nmol/min/mg, respectively. Genotyping of patients may provide a useful initial step in identifying patients who are unlikely to benefit from quinone-based chemotherapy. In the case of MMC, however, the work presented here demonstrates that genotyping of individuals with respect to NQO1 is unlikely to be beneficial in terms of predicting tumor responses to MMC.
Asunto(s)
Antibióticos Antineoplásicos/uso terapéutico , Mitomicina/uso terapéutico , NADH NADPH Oxidorreductasas/genética , Neoplasias Experimentales/tratamiento farmacológico , Animales , Modelos Animales de Enfermedad , FMN Reductasa , Genotipo , Humanos , Ratones , Ratones Desnudos , NADH NADPH Oxidorreductasas/metabolismo , Trasplante de Neoplasias , Neoplasias Experimentales/enzimología , Polimorfismo Genético , Trasplante Heterólogo , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The indolequinone EO9 demonstrated good preclinical activity but failed to show clinical efficacy against a range of tumours following intravenous drug administration. A significant factor in EO9's failure in the clinic has been attributed to its rapid pharmacokinetic elimination resulting in poor drug delivery to tumours. Intravesical administration of EO9 would circumvent the problem of drug delivery to tumours and the principal objective of this study is to determine whether or not bladder tumours have elevated levels of the enzyme NQO1 (NAD(P)H:quinone oxidoreductase) which plays a key role in activating EO9 under aerobic conditions. Elevated NQO1 levels in human bladder tumour tissue exist in a subset of patients as measured by both immunohistochemical and enzymatic assays. In a panel of human tumour cell lines, EO9 is selectively toxic towards NQO1 rich cell lines under aerobic conditions and potency can be enhanced by reducing extracellular pH. These studies suggest that a subset of bladder cancer patients exist whose tumours possess the appropriate biochemical machinery required to activate EO9. Administration of EO9 in an acidic vehicle could be employed to reduce possible systemic toxicity as any drug absorbed into the blood stream would become relatively inactive due to an increase in pH.
Asunto(s)
Antineoplásicos/uso terapéutico , Aziridinas/uso terapéutico , Indolquinonas , Indoles/uso terapéutico , Quinona Reductasas/metabolismo , Neoplasias de la Vejiga Urinaria/tratamiento farmacológico , Aziridinas/farmacocinética , Humanos , Concentración de Iones de Hidrógeno , Inmunohistoquímica , Indoles/farmacocinética , Especificidad por Sustrato , Vejiga Urinaria/enzimología , Neoplasias de la Vejiga Urinaria/enzimologíaRESUMEN
The cytotoxic activity of the imidazoacridinone C1311 was assessed on two ovarian cancer cell lines (A2780, OAW42) and one osteogenic sarcoma cell line (U2-OS) and their sublines (A2780Cp8, OAW42-MER and U2-OS-R) with experimentally induced resistance to cisplatin. A 1-h exposure to C1311 significantly inhibited the growth of all cell lines, with IC50 values ranging from 0.50 +/-0.11 to 4.10+/-0.36 microM. No or only partial cross-resistance was found between C1311 and cisplatin in the different cell lines. Treatment with equitoxic (IC50) C1311 concentrations consistently induced accumulation of cells in the G2M phase. The cyclin B1-associated p34(cdc2) kinase activity in cells arrested in G2M was superimposable to that of control cells in the OAW42-MER and U2-OS cell lines, whereas a reduction of cdc2 catalytic activity was observed in OAW42 and U2-OS-R cells. Exposure to C1311 (IC50) induced apoptosis in the U2-OS and U2-OS-R cell lines, whereas in the OAW42 and OAW42-MER cell lines there was a negligible percentage of apoptotic cells. In U2-OS, U2-OS-R and OAW42 cells, C1311 induced an increase in p53 expression and an increase in p21waf1 protein, whereas p53 failed to transactivate p21waf1 in OAW42-MER cells. An almost complete abrogation of bcl-2 was observed in U2-OS-R cells in correspondence with the peak of apoptosis induction. Our results indicate that C1311 is active against human ovarian cancer and osteogenic sarcoma cells and is not cross-resistant with CDDP. Moreover, C1311 blocks cells in the G2M phase and induces apoptosis in a small percentage of osteogenic sarcoma cells.
Asunto(s)
Aminoacridinas/farmacología , Antineoplásicos/farmacología , Neoplasias Óseas/patología , Ciclo Celular/efectos de los fármacos , Osteosarcoma/patología , Neoplasias Ováricas/patología , Apoptosis/efectos de los fármacos , División Celular/efectos de los fármacos , Ciclina B/efectos de los fármacos , Ciclina B/metabolismo , Ciclina B1 , Relación Dosis-Respuesta a Droga , Femenino , Fase G2/efectos de los fármacos , Humanos , Células Tumorales CultivadasRESUMEN
A range of 11 derivatives of flavone-8-acetic acid (FAA) in which the structure has been substantially altered in different ways have been prepared and their anti-tumour activity evaluated in vitro against a panel of human and murine tumour cell lines and in vivo against MAC 15A. The generally poor activity observed shows that the basic structure cannot be altered much without destroying the activity.
Asunto(s)
Antineoplásicos/síntesis química , Flavonoides/síntesis química , Animales , Antineoplásicos/farmacología , Flavonoides/farmacología , Humanos , Ratones , Relación Estructura-Actividad , Células Tumorales CultivadasRESUMEN
AQ4 (1,4-Bis-[[2-(dimethylamino-N-oxide)ethyl]amino]5,8-dihydroxyanthrace ne-9, 10-dione) is a prodrug designed to be excluded from cell nuclei until bioreduced in hypoxic cells to AQ4, a DNA intercalator and topoisomerase II poison. Thus, AQ4N is a highly selective bioreductive drug that is activated in, and is preferentially toxic to, hypoxic cells in tumours. Five murine tumours (MAC16, MAC26, NT, SCCVII and RIF-1) have been used to investigate the anti-tumour effects of AQ4N. In only one tumour (MAC16) was AQ4N shown to be active as a single agent. However, when combined with methods to increase the hypoxic tumour fraction in RIF-1 (by physical clamping) and MAC26 tumours (using hydralazine) there was a substantial enhancement in anti-tumour effect. Notably, RIF-1 tumours treated with AQ4N (250 mg kg(-1)) followed 15 min later by physically occluding the blood supply to the tumour for 90 min, resulted in a 13-fold increase in growth delay. When combined with radiation or chemotherapy, AQ4N substantially increased the effectiveness of these modalities in a range of in vivo model systems. AQ4N potentiates the action of radiation in both a drug and radiation dose-dependent manner. Further the enhancement observed is schedule-independent with AQ4N giving similar effects when given at any time within 16 h before or after the radiation treatment. In combination with chemotherapy it is shown that AQ4N potentiates the activity of cyclophosphamide, cisplatin and thiotepa. Both the chemotherapeutic drugs and AQ4N are given at doses which individually are close to their estimated maximum tolerated dose (data not included) which provides indirect evidence that in the combination chemotherapy experiments there is some tumour selectivity in the enhanced action of the drugs.
Asunto(s)
Antraquinonas/uso terapéutico , Antineoplásicos/uso terapéutico , Neoplasias Experimentales/tratamiento farmacológico , Neoplasias Experimentales/radioterapia , Animales , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Hipoxia de la Célula/efectos de los fármacos , Esquema de Medicación , Sinergismo Farmacológico , Masculino , Ratones , Ratones Endogámicos , Profármacos/uso terapéuticoRESUMEN
PK1 is a synthetic N-(2-hydroxypropyl)methacrylamide copolymer-doxorubicin (dox) conjugate currently undergoing Phase II evaluation in the United Kingdom. We have studied the activity of PK1 in three murine colon tumor models that differ in terms of morphology and vascularization in an attempt to determine which factors are most important in the tumor response to PK1. Vascular permeability was evaluated with Evans Blue, and pharmacokinetic studies in MAC15A and MAC26 used high-performance liquid chromatography to monitor both PK1 uptake and dox release in the tumors. Cathepsin B activity was assessed using a specific substrate. PK1 (40 mg x kg(-1) dox equivalent) was significantly more effective than dox alone (10 mg x kg(-1)) was against MAC15A tumors, which possess enhanced perfusion and retention, but not against MAC26 tumors, although MAC15A was also responsive to PK1 when grown as avascular micrometastatic deposits in the lung. Pharmacokinetic studies showed similar levels of PK1 in both tumors. Peak tumor levels of released dox were 7-fold greater in the responsive MAC15A tumor (53 microg x ml(-1)) compared with the less responsive MAC26 tumor (7.7 microg x ml(-1)) and more than 18-fold greater in MAC15A than when free dox was given. These differences in response correlated also with an increased lysosomal activity of cathepsin B. Calculated AUCs for intratumoral dox released were 431 microg x h x g(-1) and 775 microg x h x g(-1) for MAC15A and MAC26, respectively. These AUCs are 4-fold and 7-fold higher, respectively, than when dox is given alone. This study has shown that activity and the pharmacokinetics of PK1 and released dox are dependent on both the vascular properties and enzyme content of the tumors. These studies are likely to have clinical implications as aggressive tumors are known to have increased protease activity.
Asunto(s)
Antineoplásicos/farmacocinética , Neoplasias del Colon/tratamiento farmacológico , Doxorrubicina/análogos & derivados , Doxorrubicina/farmacocinética , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/secundario , Ácidos Polimetacrílicos/farmacocinética , Animales , Antineoplásicos/uso terapéutico , Área Bajo la Curva , Neoplasias del Colon/irrigación sanguínea , Neoplasias del Colon/patología , Doxorrubicina/administración & dosificación , Doxorrubicina/uso terapéutico , Portadores de Fármacos , Femenino , Neoplasias Pulmonares/irrigación sanguínea , Neoplasias Pulmonares/patología , Ratones , Ratones Endogámicos , Ácidos Polimetacrílicos/administración & dosificación , Ácidos Polimetacrílicos/uso terapéutico , Distribución TisularRESUMEN
C1311 is a novel therapeutic agent with potent activity against experimental colorectal cancer that has been selected for entry into clinical trial. The compound has previously been shown to have DNA-binding properties and to inhibit the catalytic activity of topoisomerase II. In this study, cellular uptake and mechanisms by which C1311 interacts with DNA and exerts cytotoxic effects in intact colon carcinoma cells were investigated. The HT29 colon cancer cell line was chosen to follow cellular distribution of C1311 over a time course of 24 h at drug concentrations that just inhibited cell proliferation by 50% or 100%. Nuclear uptake of C1311 and co-localization with lysosomal or mitochondrial dyes was examined by fluorescence microscopy and effects on these cellular compartments were determined by measurement of acid phosphatase levels, rhodamine 123 release or DNA-binding behaviour. The strength and mode of DNA binding was established by thermal melting stabilization, direct titration and viscometric studies of host duplex length. The onset of apoptosis was followed using a TUNEL assay and DNA-fragmentation to determine a causal relationship of cell death. Growth inhibition of HT29 cells by C1311 was concomitant with rapid drug accumulation in nuclei and in this context we showed that the compound binds to duplex DNA by intercalation, with likely A/T sequence-preferential binding. Drug uptake was also seen in lysosomes, leading to lysosomal rupture and a marked increase of acid phosphatase activity 8 h after exposure to C1311 concentrations that effect total growth inhibition. Moreover, at these concentrations lysosomal swelling and breakdown preceded apoptosis, which was not evident up to 24 h after exposure to drug. Thus, the lysosomotropic effect of C1311 appears to be a novel feature of this anticancer agent. As it is unlikely that C1311-induced DNA damage alone would be sufficient for cytotoxic activity, lysosomal rupture may be a critical component for therapeutic efficacy.
Asunto(s)
Aminoacridinas/farmacología , Antineoplásicos/farmacología , División Celular/efectos de los fármacos , Núcleo Celular/efectos de los fármacos , ADN/efectos de los fármacos , Fosfatasa Ácida/metabolismo , Aminoacridinas/farmacocinética , Antineoplásicos/farmacocinética , Apoptosis/efectos de los fármacos , Núcleo Celular/metabolismo , Neoplasias del Colon/tratamiento farmacológico , ADN/metabolismo , Fragmentación del ADN , Células HT29/efectos de los fármacos , Humanos , Microscopía FluorescenteRESUMEN
C1311 has emerged as the lead compound from a novel group of anticancer agents, the imidazoacridinones, and will be entering clinical trials shortly. Previous murine pharmacokinetic studies have shown C1311 to be rapidly and extensively distributed into tissues including tumor. This study has identified two major metabolites of C1311 and describes their pharmacokinetics in mice. M1 is a glucuronide of the parent compound with high concentrations in both plasma and liver. Calculated area under the plasma concentration versus time curve values were 6-fold and 2-fold greater, respectively, than C1311. Based on these studies, we propose M2 to be a nonfluorescent oxidation product because electrospray ionization-mass spectroscopy/mass spectroscopy analysis gave a molecular ion at m/z 367, 16 U greater than the parent compound. It formed rapidly in liver preparations in vitro, both murine and human, by a cytosolic process in the presence of NADPH and in vivo was detected in liver tissues at concentrations equivalent to those of C1311 but was not detectable in plasma. Preliminary in vitro toxicity studies showed M2 to be as potent as C1311 against MAC15A tumor cells. Over the first 24 h, 39% of the administered dose is eliminated via the bile (28%) mostly as C1311 or the kidneys (11%) as the glucuronide (M1). This study has given valuable information as to the likely metabolic pathway to occur in humans, and the cytotoxic metabolite M2 may play a role in the antitumor activity or toxicity of C1311 in the clinic.
Asunto(s)
Aminoacridinas/farmacocinética , Antineoplásicos/farmacocinética , Aminoacridinas/sangre , Aminoacridinas/orina , Animales , Antineoplásicos/sangre , Antineoplásicos/orina , Biotransformación , Cromatografía Líquida de Alta Presión , Heces/química , Femenino , Glucuronidasa/metabolismo , Semivida , Humanos , Técnicas In Vitro , Masculino , Espectrometría de Masas , Ratones , Microsomas Hepáticos/metabolismoRESUMEN
The National Cancer Institute uses the hollow fiber assay as part of its screening program for anticancer drug discovery. Angiogenesis to hollow fibers implanted s.c. has not been reported, thereby raising concerns about the efficiency of drug delivery and its subsequent effects on chemosensitivity. By extending postimplantation times beyond the 6-day period presently used, extensive vascular networks develop, resulting in both increased delivery and chemosensitivity to doxorubicin. This study suggests that present protocols used to evaluate compounds may produce false negative results, and additional studies to determine the predictive value of the assay are required.
Asunto(s)
Adenocarcinoma/irrigación sanguínea , Adenocarcinoma/tratamiento farmacológico , Neoplasias del Colon/irrigación sanguínea , Neoplasias del Colon/tratamiento farmacológico , Ensayos de Selección de Medicamentos Antitumorales/métodos , Neovascularización Patológica/metabolismo , Adenocarcinoma/patología , Animales , Antibióticos Antineoplásicos/farmacocinética , Antibióticos Antineoplásicos/farmacología , Antineoplásicos/farmacocinética , Antineoplásicos/farmacología , Neoplasias del Colon/patología , Modelos Animales de Enfermedad , Doxorrubicina/farmacocinética , Doxorrubicina/farmacología , Inyecciones Subcutáneas , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Neovascularización Patológica/patología , Células Tumorales CultivadasRESUMEN
C1311 is the most active member of a new series of rationally designed anti-cancer agents, the imidazoacridinones, which has shown promising pre-clinical anti-tumour activity in vitro and in vivo against a variety of human colon cancers and is a strong candidate for clinical trials. Data are not available on the pharmacokinetic properties of this compound; therefore, the main aim of this project was to study the plasma pharmacokinetics and tissue and tumour distribution of C1311 in mice and to assess, prior to potential clinical application, whether these pharmacokinetics were linear with respect to the dose. The distribution of C1311 in whole blood was also studied. NMRI or NCR-Nu mice were used throughout the study. C1311 was given i.p. at doses of 15, 50, 100 and (the maximum tolerated dose, (MTD) 150 mg kg(-l) i.p. Plasma, tissue and tumour levels were monitored over a 24-h period using high-performance liquid chromatography (HPLC) with fluorescence detection. The distribution of C1311 in murine and human whole blood was studied using both HPLC and fluorescence microscopy. C1311 was quickly cleared from the plasma (47410 ml min kg(-1)) and rapidly distributed into the tissues at all doses. Tissue-to-plasma ratios were large, ranging from 8 in the liver (15 mg kg(-l)) to 600 (50 mg kg(-1)) in the spleen. Overall concentrations were ranked in the order of plasma << liver < kidney < fat < small intestine < spleen. Tumour concentrations were similar to those measured in the liver and kidney, with AUCs being 186 (MAC15A) and 94.4 microg h ml(-l)(HT-29). Plasma pharmacokinetics were linear at doses of 15-100 mg kg(-1), but disproportionate increases were seen in plasma and tissue concentrations at doses above 100 mg kg(-l). C1311 distributed unevenly in both mouse and human blood, with higher concentrations occurring in the cellular fraction than in plasma. Nucleated cells accounted for a large proportion of this localised drug. In conclusion, C1311 is quickly cleared from the plasma and rapidly distributed into the tissues, with tissue concentrations being far higher than plasma levels. The plasma pharmacokinetics are linear up to but not above doses of 100 mg kg(-1). Concentrations of C1311 are greater in the cellular fraction of the blood than in the plasma, with disproportionately high concentrations occurring in the nucleated fraction.
Asunto(s)
Aminoacridinas/farmacocinética , Antineoplásicos/farmacocinética , Tejido Adiposo/metabolismo , Aminoacridinas/administración & dosificación , Aminoacridinas/sangre , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/sangre , Área Bajo la Curva , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Inyecciones Intraperitoneales , Riñón/metabolismo , Hígado/metabolismo , Ratones , Bazo/metabolismoRESUMEN
Loss of DNA mismatch repair has been described in a number of tumour types such as colorectal adenocarcinoma and leads to microsatellite instability. This may have clinical relevance due to mismatch repair defects altering chemosensitivity towards certain classes of anti-tumour agent. This study has examined microsatellite instability of eight murine colon adenocarcinoma tumour models induced by 1,2-dimethylhydrazine. Four microsatellite regions were examined suggesting that four of the tumour models exhibit a low level of microsatellite instability. Loss of heterozygosity was found in 5/8 tumours, suggesting that allelic loss may be a relatively common step in the carcinogenesis of these tumour models. Three of the allelic losses involved the D11MIT4 locus which is situated very close to the p53 tumour suppressor locus. Four tumour models are routinely cultured in vitro and these were used to examine whether there was any association between microsatellite instability, mutant frequency and chemo-sensitivity of these tumour models, comparing them with four human adenocarcinoma cell lines of known mismatch repair status. Two cell lines (MAC26 and MAC16) were found to be more chemoresistant towards cisplatin but not 6-thioguanine. No association was found between microsatellite instability and chemosensitivity for either the human or mouse cell lines.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/genética , Neoplasias del Colon/tratamiento farmacológico , Neoplasias del Colon/genética , Repeticiones de Microsatélite , Mutación , 1,2-Dimetilhidrazina , Adenocarcinoma/inducido químicamente , Animales , Antimetabolitos Antineoplásicos/farmacología , Antineoplásicos/farmacología , Carcinógenos , Cisplatino/farmacología , Neoplasias del Colon/inducido químicamente , Reparación del ADN , ADN de Neoplasias/genética , Modelos Animales de Enfermedad , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Pérdida de Heterocigocidad , Ratones , Tioguanina/farmacología , Células Tumorales CultivadasRESUMEN
A range of 14 derivatives of flavone-8-acetic acid (FAA) with a heterocyclic substituent in place of the 2-phenyl group have been prepared and their anti-tumour activity evaluated in vitro against a panel of human and murine tumour cell lines and in vivo against MAC 15A. Some of the compounds, notably 2c,d and s, showed significant in vivo activity and these require further studies in order to evaluate their potential for development.
Asunto(s)
Antineoplásicos/síntesis química , Flavonoides/síntesis química , Flavonoides/farmacología , Compuestos Heterocíclicos/síntesis química , Animales , Antineoplásicos/farmacología , Compuestos Heterocíclicos/farmacología , Humanos , Ratones , Estructura Molecular , Neoplasias Experimentales/tratamiento farmacológico , Células Tumorales CultivadasRESUMEN
A range of 18 derivatives of flavone-8-acetic acid (FAA) with substituents on the 2-phenyl group have been prepared and their anti-tumour activity evaluated in vitro against a panel of human and murine tumour cell lines and in vivo against MAC 15A. There was no clear-cut relationship between in vitro and in vivo activity but the activity in each situation was found to be very sensitive to the precise substitution pattern with closely related isomers giving widely different activities. Some of the compounds, notably 10b,c,j, and r, were active in vivo and these require further studies in order to evaluate their potential for development.
Asunto(s)
Antineoplásicos/síntesis química , Antineoplásicos/farmacología , Flavonoides/síntesis química , Flavonoides/farmacología , Animales , Neoplasias del Colon/tratamiento farmacológico , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Ratones , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Human breast cancer cell lines are required as models for use in the understanding of breast carcinoma, and for improving the ability of cell screens to detect appropriate anti-cancer agents. Four human breast cancer cell lines (MT-1, MaTu. MT-3 and MC4000) were established from human tumour xenografts grown in nude mice. All the lines were shown to be of human origin by karyotype analysis, were epithelial in morphology by both light and electron microscopy, were positive for cytokeratin 18, and were free from mycoplasma, bacterial, yeast and fungal contamination. All of the new lines were shown to be ER and PgR negative, while using the same procedures (i.e. radioligand binding and immunohistochemical staining) the positive control cell line MCF-7 was shown to be positive. MaTu had been previously reported as ER and PgR positive in vivo and it may be that this characteristic had been lost due to in vitro selection pressures. The growth rates of all the new breast cancer cell lines were similar and within the limits required for incorporation into a panel for screening anti-cancer drugs by a microtetrazolium based, colorimetric growth inhibition assay. Three of the lines (MT-1. MaTu and MC4000) were also able to grow into macroscopic colonies for use in a non-agar clonogenic assay. In addition, both MT-1 and MaTu formed spheroids and were clonogenic in soft-agar. The new lines demonstrated a wide range of sensitivities to anticancer agents commonly used in the treatment of breast cancer, and together with their corresponding xenografts are providing additional systems for the evaluation of new compounds.
Asunto(s)
Antineoplásicos/toxicidad , Neoplasias de la Mama , Animales , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Neoplasias de la Mama/ultraestructura , Técnicas de Cultivo de Célula/métodos , División Celular , Línea Celular , Supervivencia Celular/efectos de los fármacos , Mapeo Cromosómico , Cromosomas Humanos , Femenino , Humanos , Cariotipificación , Queratinas/análisis , Metalotioneína 3 , Ratones , Ratones Desnudos , Microscopía Electrónica , Mycoplasma/aislamiento & purificación , Ploidias , Ensayo de Unión Radioligante , Receptores de Estrógenos/análisis , Receptores de Progesterona/análisis , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
Telomerase, a ribonucleoprotein, elongates and/or maintains telomeres by adding TTAGGG tandem repeat sequences using the RNA component of the enzyme as a template. Enzyme activity appears to be associated with cell immortalisation and malignant progression as telomerase activity has been found in the majority of human tumours, but not in most somatic cells or tissues. Telomerase inhibition has, therefore, been proposed as a novel and potentially selective target for therapeutic intervention. Since telomeric tandem repeats as well as the human telomerase RNA component (hTR) and its gene are guanosine-rich, we examined whether the sequence specific, G-Pt-G, cross-linking agent cisplatin is capable of inhibiting telomerase activity. The TRAP assay was used to measure telomerase activity in cisplatin treated cell extracts and RT-PCR strategies used to examine hTR expression after drug exposure. Cisplatin reduced telomerase activity in a specific and concentration-dependent manner in human testicular tumour cells, whilst doxorubicin, bleomycin, methotrexate, melphalan and transplatin had no effect. It is proposed that telomerase inhibition might be a component of the efficacy of cisplatin in the treatment of testicular cancer.