RESUMO
Cancer incidence is rising and this global challenge is further exacerbated by tumour resistance to available medicines. A promising approach to meet the need for improved cancer treatment is drug repurposing. Here we highlight the potential for repurposing disulfiram (also known by the trade name Antabuse), an old alcohol-aversion drug that has been shown to be effective against diverse cancer types in preclinical studies. Our nationwide epidemiological study reveals that patients who continuously used disulfiram have a lower risk of death from cancer compared to those who stopped using the drug at their diagnosis. Moreover, we identify the ditiocarb-copper complex as the metabolite of disulfiram that is responsible for its anti-cancer effects, and provide methods to detect preferential accumulation of the complex in tumours and candidate biomarkers to analyse its effect on cells and tissues. Finally, our functional and biophysical analyses reveal the molecular target of disulfiram's tumour-suppressing effects as NPL4, an adaptor of p97 (also known as VCP) segregase, which is essential for the turnover of proteins involved in multiple regulatory and stress-response pathways in cells.
Assuntos
Dissuasores de Álcool , Alcoolismo/tratamento farmacológico , Antineoplásicos , Dissulfiram/farmacologia , Dissulfiram/uso terapêutico , Reposicionamento de Medicamentos , Neoplasias/tratamento farmacológico , Proteínas Nucleares/metabolismo , Adulto , Dissuasores de Álcool/farmacologia , Dissuasores de Álcool/uso terapêutico , Alcoolismo/epidemiologia , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Cobre/química , Dinamarca/epidemiologia , Dissulfiram/química , Feminino , Resposta ao Choque Térmico/efeitos dos fármacos , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Terapia de Alvo Molecular , Neoplasias/metabolismo , Neoplasias/mortalidade , Neoplasias/patologia , Proteínas Nucleares/química , Agregados Proteicos , Ligação Proteica/efeitos dos fármacos , Proteólise/efeitos dos fármacosRESUMO
Wilson's disease is a genetic disorder that causes excessive accumulation of copper in the body, leading to toxic damage, especially in the liver and nervous system. The current treatment cause burdensome side effects. We describe the use of chemically modified biopolymer carriers based on microcrystalline cellulose and chitosan containing the highly specific copper chelator 8-hydroxyquinoline as a new type of therapy for Wilson's disease. The chelators can scavenges copper ions released from food during digestion and copper ions present in secretions in the gastrointestinal tract. Because the chelator is covalently bound to indigestible biopolymer carriers (crosslinked chitosan or modified cellulose), it is not taken up by the gastrointestinal tract and it can be eliminated through the feces, avoiding unwanted side effects. This concept was tested on Wistar rats, which received a radioactive 64CuCl2 solution together with the polymers with covalently bound 8-hydroxyquinoline through a gastric probe. 64Copper complex uptake from the gastrointestinal tract was significantly inhibited by both chelating polymers. With the modified polymers, the presence of 64Cu was detected mostly in the gastrointestinal tract, not in the internal organs. These findings indicate modified cellulose and crosslinked chitosan, with covalently bound 8-hydroxyquinoline exhibited the potential to be excellent therapeutics for treating Wilson's disease.
Assuntos
Celulose/administração & dosagem , Quitosana/administração & dosagem , Degeneração Hepatolenticular/tratamento farmacológico , Oxiquinolina/administração & dosagem , Animais , Celulose/farmacocinética , Quitosana/farmacocinética , Cobre , Radioisótopos de Cobre/administração & dosagem , Radioisótopos de Cobre/farmacocinética , Trato Gastrointestinal/metabolismo , Degeneração Hepatolenticular/metabolismo , Oxiquinolina/farmacocinética , Ratos WistarRESUMO
We designed and synthesized a new delivery system for the anticancer drug doxorubicin based on a biocompatible hydrophilic poly(2-ethyl-2-oxazoline) (PEtOx) carrier with linear architecture and narrow molar mass distribution. The drug is connected to the polymer backbone via an acid-sensitive hydrazone linker, which allows its triggered release in the tumor. The in vitro studies demonstrate successful cellular uptake of conjugates followed by release of the cytostatic cargo. In vivo experiments in EL4 lymphoma bearing mice revealed prolonged blood circulation, increased tumor accumulation and enhanced antitumor efficacy of the PEtOx conjugate having higher molecular weight (40 kDa) compared to the lower molecular weight (20 kDa) polymer. Finally, the in vitro and in vivo anti-cancer properties of the prepared PEtOx conjugates were critically compared with those of the analogous system based on the well-established PHPMA carrier. Despite the relatively slower intracellular uptake of PEtOx conjugates, resulting also in their lower cytotoxicity, there are no substantial differences in in vivo biodistribution and anti-cancer efficacy of both classes of polymer-Dox conjugates. Considering the synthetic advantages of poly(2-alkyl-2-oxazoline)s, the presented study demonstrates their potential as a versatile alternative to well-known PEO- or PHPMA-based materials for construction of drug delivery systems.
Assuntos
Acrilamidas/química , Doxorrubicina/química , Doxorrubicina/uso terapêutico , Portadores de Fármacos/química , Nanomedicina/métodos , Poliaminas/química , Polímeros/química , Animais , Linhagem Celular Tumoral , Feminino , Citometria de Fluxo , Células HeLa , Humanos , Células MCF-7 , Camundongos , Camundongos Endogâmicos C57BL , Microscopia ConfocalRESUMO
PURPOSE: To determine the rate of opacification of hydrophilic MemoryLens U940A intraocular lenses (IOLs) (Mentor Ophthalmics, Inc.) in the given cohort and perform a histopathological and spectrophotometer analysis of 2 explanted opacified IOLs. SETTING: Ophthalmology Department, Faculty Hospital, Nitra, Slovakia. METHOD: This retrospective study comprised 182 patients (205 eyes) who had implantation of a MemoryLens U940A IOL from June 1997 to June 2000. The patients were examined using a slitlamp to detect the presence of IOL opacification. In 4 cases, the lenses were explanted because of significant opacification and patient-reported problems; 2 lenses were provided for further analysis. One unused reference MemoryLens U940A IOL was also evaluated. All IOL were stained with von Kossa to determine the presence of calcium in the opacification. To confirm the components presence of an ultraviolet (UV) absorber, the IOLs were examined with an Avatar 330 Fourier transfer infrared (IR) spectroscope and a UV visible spectrophotometer (Philips). The IR spectrums for the IOL were identified using an IR spectrum atlas. The opacified IOLs, reference IOL, and the IOL packaging were further examined to determine the presence of silicone. RESULTS: Various amounts of opacification were found on the MemoryLens U940A IOL in 30 eyes (30 patients) (14.63%). Two explanted IOLs were positive for von Kossa staining, proving the presence of calcium deposits; the reference lens staining was negative. Spectrophotometry showed that the reference IOL and opacified IOLs were of the same polymer. The presence of the UV absorber on the benzophenone base was seen in the reference lens but not the opacified IOLs. In contrast, an increased concentration of low-molecular-weight components generated during the degradation of the polymer was present in the opacified lenses. The white cover pf the IOL is of polydimethyl siloxane, a silicone rubber. However, no silicone rubber was present in any examined lens, perhaps because the IOLs were in contact with alcohol during the histopathologic examination. CONCLUSIONS: The results indicate opacification of the hydrophilic MemoryLens U940A was caused by premature consumption of the UV absorber in the polymer component of the IOLs optic, with a subsequent degradation of the polymer. Whether silicone from the white cover led to the IOL opacification, as reported with other types of hydrophilic IOLs, could not be confirmed.
Assuntos
Materiais Biocompatíveis/química , Lentes Intraoculares , Falha de Prótese , Idoso , Cálcio/análise , Remoção de Dispositivo , Feminino , Humanos , Implante de Lente Intraocular , Facoemulsificação , Estudos Retrospectivos , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Auger electrons-emitting radioisotopes (such as iodine-125) are a potentially effective cancer treatment. They are extremely biologically effective, but only within a short range (nanometers). Their use as an effective cancer therapy requires that they will be transported within close proximity of DNA by an intercalator, where they induce double-strand breaks leading to cell death. This type of therapy may be even more beneficial when associated with drug delivery systems. In this report, we describe an optimized triple-targeted polymer delivery system for the intercalator ellipticine, which contains radioisotope iodine-125 with high specific radioactivity (63.2 GBq/mg). This compound is linked to an N-(2-hydroxypropyl)methacrylamide copolymer via an optimized acid-sensitive hydrazone linker. The system is stable at pH 7.4 (representing the pH of blood plasma), and the radioiodine-containing biologically active intercalator is released upon a decrease in pH (44% of the intercalator is released after 24h of incubation in pH 5.0 buffer, which mimics the pH in late endosomes). The active compound is a potent intercalator, as shown with direct titration with a DNA solution, and readily penetrates into cell nuclei, as observed by confocal microscopy. Its polymer conjugate is internalized into endosomes and releases the radioactive intercalator, which accumulates in the cell nuclei. In vivo experiments on mice with 4T1 murine breast cancer resulted in a statistically significant increase in the survival of mice treated with the polymer radioconjugate. The free radiolabeled intercalator was also shown to be effective, but it was less potent than the polymer conjugate.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Elétrons , Elipticinas/farmacologia , Polímeros/química , Animais , Antineoplásicos/química , Neoplasias da Mama/patologia , Sistemas de Liberação de Medicamentos , Ensaios de Seleção de Medicamentos Antitumorais , Elipticinas/química , Feminino , Concentração de Íons de Hidrogênio , Radioisótopos do Iodo , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Molecular , Polímeros/síntese química , Relação Estrutura-AtividadeRESUMO
Wilson's disease is a genetic disorder caused by a malfunction of ATPase 7B that leads to high accumulation of copper in the organism and consequent toxic effects. We propose a gentle therapy to eliminate the excessive copper content with oral administration of insoluble non-resorbable polymer sorbents containing selective chelating groups for copper(II). Polymeric beads with the chelating agents triethylenetetramine, N,N-di(2-pyridylmethyl)amine, and 8-hydroxyquinoline (8HQB) were investigated. In a preliminary copper uptake experiment, we found that 8HQB significantly reduced copper uptake (using copper-64 as a radiotracer) after oral administration in Wistar rats. Furthermore, we measured organ radioactivity in rats to demonstrate that 8HQB radiolabelled with iodine-125 is not absorbed from the gastrointestinal tract after oral administration. Non-resorbability and the blockade of copper uptake were also confirmed with small animal imaging (PET/CT) in mice. In a long-term experiment with Wistar rats fed a diet containing the polymers, we have found that there were no signs of polymer toxicity and the addition of polymers to the diet led to a significant reduction in the copper contents in the kidneys, brains, and livers of the rats. We have shown that polymers containing specific ligands could potentially be novel therapeutics for Wilson's disease.
Assuntos
Quelantes/uso terapêutico , Cobre/metabolismo , Degeneração Hepatolenticular/tratamento farmacológico , Metilmetacrilatos/uso terapêutico , Administração Oral , Animais , Encéfalo/metabolismo , Quelantes/química , Feminino , Trato Gastrointestinal/metabolismo , Degeneração Hepatolenticular/metabolismo , Rim/metabolismo , Fígado/metabolismo , Metilaminas/química , Metilaminas/uso terapêutico , Metilmetacrilatos/química , Camundongos , Oxiquinolina/química , Oxiquinolina/uso terapêutico , Piridinas/química , Piridinas/uso terapêutico , Ratos Wistar , Trientina/química , Trientina/uso terapêuticoRESUMO
BACKGROUND: Clinically-approved anticancer photodynamic therapy (PDT) is now extensively studied for various cancer diagnoses. We focused on the treatment efficacy of topical administration of hydroxy-aluminum phthalocyanine (AlOH-PC) entrapped in liposomes against in vivo models of prostate carcinomas. MATERIALS AND METHODS: LNCaP and PC3 cells were subcutaneously injected into the right flank of athymic nude mice. Mice with grown tumours were used for in vivo efficacy studies. Firstly, we applied different doses of AlOH-PC to less aggressive LNCaP tumours to determine the effective dose. In later studies, we focused on more aggressive prostate tumours (PC3) using doses of liposomal-AlOH-PC gel formulation. Topical application of photosensitizers was followed by PDT irradiation (600-700 nm, 635 nm peak). Tumour growth was measured three times-a-week. RESULTS: Comparison of PDT of aggressive PC3 and less aggressive LNCaP prostate carcinomas showed that both tumour types are sensitive and treatable by liposomal formulation of AlOH-PC. For LNCaP tumours the efficient dose (100% experimental animals cured, n=8/8) was 4.5 mg/ml of AlOH-PC in the gel. Whereas, in the case of PC3 carcinomas, a dose of 4 mg/ml significantly postponed tumour growth, but no animals were cured (n=0/8); a sufficient curative dose (100% mice cured, n=8/8) was 6 mg/ml of AlOH-PC in the gel. CONCLUSION: Liposomal AlOH-PC gel has potential for effective PDT of prostate carcinomas.