RESUMO
Pseudomonas aeruginosa possesses an unusual pathway for 5'-methylthioadenosine (MTA) metabolism involving deamination to 5'-methylthioinosine (MTI) followed by N-ribosyl phosphorolysis to hypoxanthine and 5-methylthio-α-d-ribose 1-phosphate. The specific MTI phosphorylase of P. aeruginosa has been reported [Guan, R., Ho, M. C., Almo, S. C., and Schramm, V. L. (2011) Biochemistry 50, 1247-1254], and here we characterize MTA deaminase from P. aeruginosa (PaMTADA). Genomic analysis indicated the PA3170 locus to be a candidate for MTA deaminase (MTADA). Protein encoded by PA3170 was expressed and shown to deaminate MTA with 40-fold greater catalytic efficiency for MTA than for adenosine. The k(cat)/K(m) value of 1.6 × 10(7) M(-1) s(-1) for MTA is the highest catalytic efficiency known for an MTA deaminase. 5'-Methylthiocoformycin (MTCF) is a 4.8 pM transition state analogue for PaMTADA but causes no significant inhibition of human adenosine deaminase or MTA phosphorylase. MTCF is permeable to P. aeruginosa and exhibits an IC(50) of 3 nM on cellular PaMTADA activity. PaMTADA is the only activity in P. aeruginosa extracts to act on MTA. MTA and 5-methylthio-α-d-ribose are involved in quorum sensing pathways; thus, PaMTADA is a potential target for quorum sensing. The crystal structure of PaMTADA in complex with MTCF shows the transition state mimic 8(R)-hydroxyl group in contact with a catalytic site Zn(2+), the 5'-methylthio group in a hydrophobic pocket, and the transition state mimic of the diazepine ring in contact with a catalytic site Glu.
Assuntos
Desoxiadenosinas/metabolismo , Nucleosídeo Desaminases/metabolismo , Pseudomonas aeruginosa/enzimologia , Percepção de Quorum , Tionucleosídeos/metabolismo , Adenosina Desaminase/metabolismo , Sequência de Aminoácidos , Coformicina/análogos & derivados , Coformicina/farmacologia , Cristalografia por Raios X , Humanos , Ligases , Metiltioinosina/metabolismo , Modelos Moleculares , Dados de Sequência Molecular , Nucleosídeo Desaminases/antagonistas & inibidores , Alinhamento de Sequência , Especificidade por SubstratoRESUMO
The effects of adenosine were studied on human neutrophils with respect to their generation of superoxide anion, degranulation, and aggregation in response to soluble stimuli. Adenosine markedly inhibited superoxide anion generation by neutrophils stimulated with N-formyl methionyl leucyl phenylalanine (FMLP), concanavalin A (Con A), calcium ionophore A23187, and zymosan-treated serum; it inhibited this response to PMA to a far lesser extent. The effects of adenosine were evident at concentrations ranging from 1 to 1,000 microM with maximal inhibition at 100 microM. Cellular uptake of adenosine was not required for adenosine-induced inhibition since inhibition was maintained despite the addition of dipyridamole, which blocks nucleoside uptake. Nor was metabolism of adenosine required, since both deoxycoformycin (DCF) and erythro-9-(2-hydroxy-3-nonyl) adenine did not interfere with adenosine inhibition of superoxide anion generation. The finding that 2-chloroadenosine, which is not metabolized, resembled adenosine in its ability to inhibit superoxide anion generation added further evidence that adenosine metabolism was not required for inhibition of superoxide anion generation by neutrophils. Unexpectedly, endogenously generated adenosine was present in supernatants of neutrophil suspensions at 0.14-0.28 microM. Removal of endogenous adenosine by incubation of neutrophils with exogenous adenosine deaminase (ADA) led to marked enhancement of superoxide anion generation in response to FMLP. Inactivation of ADA with DCF abrogated the enhancement of superoxide anion generation. Thus, the enhancement was not due to a nonspecific effect of added protein. Nor was the enhancement due to the generation of hypoxanthine or inosine by deamination of adenosine, since addition of these compounds did not affect neutrophil function. Adenosine did not significantly affect either aggregation or lysozyme release and only modestly affected beta-glucuronidase release by neutrophils stimulated with FMLP. These data indicate that adenosine (at concentrations that are present in plasma) acting via cell surface receptors is a specific modulator of superoxide anion generation by neutrophils.
Assuntos
Adenosina/fisiologia , Neutrófilos/metabolismo , Superóxidos/sangue , 2-Cloroadenosina , Adenina/análogos & derivados , Adenina/farmacologia , Adenosina/análogos & derivados , Adenosina/sangue , Adenosina/farmacologia , Adenosina Desaminase/farmacologia , Calcimicina/farmacologia , Agregação Celular/efeitos dos fármacos , Coformicina/análogos & derivados , Coformicina/farmacologia , Citocalasina B/farmacologia , Grânulos Citoplasmáticos/enzimologia , Depressão Química , Dipiridamol/farmacologia , Humanos , Metilação , N-Formilmetionina Leucil-Fenilalanina/farmacologia , PentostatinaRESUMO
Plasmodium falciparum is a purine auxotroph requiring hypoxanthine as a key metabolic precursor. Erythrocyte adenine nucleotides are the source of the purine precursors, making adenosine deaminase (ADA) a key enzyme in the pathway of hypoxanthine formation. Methylthioadenosine (MTA) is a substrate for most malarial ADAs, but not for human ADA. The catalytic site specificity of malarial ADAs permits methylthiocoformycin (MT-coformycin) to act as a Plasmodium-specific transition state analogue with low affinity for human ADA [Tyler, P. C., Taylor, E. A., Frohlich, R. G. G., and Schramm, V. L. (2007) J. Am. Chem. Soc. 129, 6872-6879]. The structural basis for MTA and MT-coformycin specificity in malarial ADAs is the subject of speculation [Larson, E. T., et al. (2008) J. Mol. Biol. 381, 975-988]. Here, the crystal structure of ADA from Plasmodium vivax (PvADA) in a complex with MT-coformycin reveals an unprecedented binding geometry for 5'-methylthioribosyl groups in the malarial ADAs. Compared to malarial ADA complexes with adenosine or deoxycoformycin, 5'-methylthioribosyl groups are rotated 130 degrees . A hydrogen bonding network between Asp172 and the 3'-hydroxyl of MT-coformycin is essential for recognition of the 5'-methylthioribosyl group. Water occupies the 5'-hydroxyl binding site when MT-coformycin is bound. Mutagenesis of Asp172 destroys the substrate specificity for MTA and MT-coformycin. Kinetic, mutagenic, and structural analyses of PvADA and kinetic analysis of five other Plasmodium ADAs establish the unique structural basis for its specificity for MTA and MT-coformycin. Plasmodium gallinaceum ADA does not use MTA as a substrate, is not inhibited by MT-coformycin, and is missing Asp172. Treatment of P. falciparum cultures with coformycin or MT-coformycin in the presence of MTA is effective in inhibiting parasite growth.
Assuntos
Adenosina Desaminase/metabolismo , Coformicina/análogos & derivados , Coformicina/química , Coformicina/metabolismo , Malária Falciparum/enzimologia , Plasmodium falciparum/enzimologia , Adenosina Desaminase/química , Animais , Antimaláricos/química , Antimaláricos/metabolismo , Antimaláricos/farmacologia , Coformicina/farmacologia , Humanos , Malária Falciparum/metabolismo , Malária Falciparum/prevenção & controle , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/crescimento & desenvolvimento , Relação Estrutura-Atividade , Especificidade por SubstratoRESUMO
The concentrations of renal ATP have been measured by 31P-nuclear magnetic resonance (NMR) before, during, and after bilateral renal artery occlusion. Using in vivo NMR, the initial postischemic recovery of ATP increased with the magnitude of the residual nucleotide pool at the end of ischemia. ATP levels after 120 min of reflow correlated with functional recovery at 24 h. In the present study the effect of blocking the degradation of ATP during ischemia upon the postischemic restoration of ATP was investigated. Inhibition of adenosine deaminase by 80% with the tight-binding inhibitor 2'-deoxycoformycin led to a 20% increase in the residual adenine nucleotide pool. This increased the ATP initial recovery after 45 min of ischemia from 52% (in controls) to 62% (in the treated animals), as compared to the basal levels. The inhibition also caused an accelerated postischemic restoration of cellular ATP so that at 120 min it was 83% in treated rats vs. 63% in untreated animals. There was a corresponding improvement in the functional recovery from the insult (increase of 33% in inulin clearance 24 h after the injury). Inhibition of adenosine deaminase during ischemia results in a injury similar to that seen after a shorter period of insult.
Assuntos
Inibidores de Adenosina Desaminase , Rim/irrigação sanguínea , Nucleosídeo Desaminases/antagonistas & inibidores , Trifosfato de Adenosina/análise , Animais , Coformicina/análogos & derivados , Coformicina/farmacologia , Inulina/farmacocinética , Isquemia , Espectroscopia de Ressonância Magnética , Masculino , Pentostatina , Ratos , Ratos EndogâmicosRESUMO
To delineate the extent to which bone marrow transplantation provides "enzyme replacement therapy", we have determined metabolite concentrations in two patients with adenosine deaminase (ADA) deficiency treated with bone marrow transplants and rendered immunologically normal. 10 yr after engraftment of lymphoid cells, erythrocyte deoxy ATP was markedly decreased compared to the marked elevations of deoxy ATP observed in untreated patients, but was still significantly elevated (62 and 90 vs. normal of 6.0 +/- 6.0 nmol/ml packed erythrocytes). Similarly, deoxyadenosine and adenosine excretion were both markedly diminished compared to that of untreated patients but deoxyadenosine excretion was still clearly increased (20.1 and 38.6 vs. normal of less than 0.2 nmol/mg creatinine) while adenosine excretion was in the upper range of normal (7.0 and 8.1 vs. normal of 5.6 +/- 3.6 nmol/mg creatinine). Mononuclear cell deoxy ATP content was also elevated compared to normal (5.25 and 14.4 vs. 1.2 +/- 0.3). Separated mononuclear cells of bone marrow transplanted patients contain both donor lymphocytes and recipient monocytes. When mononuclear cells were depleted of the cells enriched for donor lymphocytes (i.e. monocyte depleted) was lower than that of the mixed mononuclear cells (2.2 vs. 5.26). Surprisingly, plasma adenosine was as high as in untreated ADA-deficient patients (3.2 and 1.5 vs. untreated of 0.3-3 microM). Consistent with the elevated plasma adenosine and urinary deoxyadenosine, erythrocyte S-adenosyl homocysteine hydrolase activity was diminished (0.88 and 1.02 vs. normal of 5.64 +/- 0.25). Thus, bone marrow transplantation of ADA-deficient patients not only provides lymphoid stem cells, but also partially, albeit incompletely, clears abnormally increased metabolites from nonlymphoid body compartments.
Assuntos
Adenosina Desaminase/deficiência , Transplante de Medula Óssea , Nucleosídeo Desaminases/deficiência , Purinas/metabolismo , Adenosina/sangue , Adenosina/farmacologia , Adenosina/urina , Adenosil-Homocisteinase , Criança , Coformicina/análogos & derivados , Coformicina/farmacologia , Nucleotídeos de Desoxiadenina/metabolismo , Desoxiadenosinas/sangue , Desoxiadenosinas/urina , Eritrócitos/metabolismo , Seguimentos , Humanos , Hidrolases/metabolismo , Monócitos/metabolismo , PentostatinaRESUMO
Myocardial ischemia is characterized by the liberation of adenosine and by complement-mediated inflammation. We have reported that amidated C3, formed when ammonia (NH3) disrupts the thiolester bond of C3, serves as an alternative pathway convertase, generates C5b-9, and stimulates phagocytic oxidative metabolism. We investigated whether the deamination of adenosine by adenosine deaminase in hematopoietic cells might liberate sufficient ammonia to form amidated C3 and thereby trigger complement-mediated inflammation at ischemic sites. In the presence of 4 mM adenosine, NH3 production per erythrocyte (RBC) was equal to that per neutrophil (PMN) (3.3 X 10(-15) mol/cell per h). Because RBC outnumber PMN in normal blood by a thousandfold, RBC are the major source of NH3 production in the presence of adenosine. NH3 production derived only from the deamination of adenosine by the enzyme adenosine deaminase and was abolished by 0.4 microM 2'-deoxycoformycin, a specific inhibitor of adenosine deaminase. When purified human C3 was incubated with 5 X 10(8) human RBC in the presence of adenosine, disruption of the C3 thiolester increased more than twofold over that measured in C3 incubated with buffer, or in C3 incubated with RBC (P less than 0.05). The formation of amidated C3 was abolished by the preincubation of RBC with 2'-deoxycoformycin (P less than 0.001). Amidated C3 elicited statistically significant release of superoxide, myeloperoxidase, and lactoferrin from PMN. Thus, the formation of amidated C3 by RBC deamination of adenosine triggers a cascade of complement-mediated inflammatory reactions.
Assuntos
Adenosina Desaminase/fisiologia , Complemento C3/metabolismo , Eritrócitos/metabolismo , Inflamação/etiologia , Nucleosídeo Desaminases/fisiologia , Adenosina/metabolismo , Adulto , Amônia/metabolismo , Coformicina/análogos & derivados , Coformicina/farmacologia , Feminino , Humanos , Masculino , PentostatinaRESUMO
Deoxyadenosine has been implicated as the toxic metabolite causing profound lymphopenia in immunodeficient children with a genetic deficiency of adenosine deaminase (ADA), and in adults treated with the potent ADA inhibitor deoxycoformycin. However, the biochemical basis for deoxyadenosine toxicity toward lymphocytes remains controversial. The present experiments have examined in detail the sequential metabolic changes induced in nondividing human peripheral blood lymphocytes by incubation with deoxyadenosine plus deoxycoformycin, or with 2-chlorodeoxyadenosine (CdA), an ADA resistant deoxyadenosine congener with anti-leukemic and immunosuppressive properties. The lymphotoxic effect of deoxyadenosine and CdA required their phosphorylation, and was inhibited by deoxycytidine. As early as 4 h after exposure to the deoxynucleosides, strand breaks in lymphocyte DNA began to accumulate, and RNA synthesis decreased. These changes were followed by a significant fall in intracellular NAD levels at 8 h, a drop in ATP pools at 24 h, and cell death by 48 h. Incubation of the lymphocytes with 5 mM nicotinamide, a NAD precursor and an inhibitor of poly(ADP-ribose) synthetase, prevented NAD depletion. The nicotinamide treatment also rendered the lymphocytes highly resistant to deoxyadenosine and CdA toxicity, without altering dATP formation or the accumulation of DNA strand breaks. The poly(ADP-ribose) synthetase inhibitor 3-aminobenzamide exerted a similar although less potent effect. These results suggest that NAD depletion, probably triggered by poly(ADP-ribose) formation, is the principle cause of death in normal resting human lymphocytes exposed to deoxyadenosine plus deoxycoformycin, or to CdA.
Assuntos
Desoxiadenosinas/análogos & derivados , Desoxiadenosinas/farmacologia , Linfócitos/efeitos dos fármacos , Trifosfato de Adenosina/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Cladribina , Coformicina/análogos & derivados , Coformicina/farmacologia , DNA/metabolismo , Humanos , Técnicas In Vitro , Linfócitos/metabolismo , NAD/metabolismo , Niacinamida/farmacologia , Pentostatina , RNA/biossínteseRESUMO
2'-Deoxycoformycin (DCF) is an inhibitor of the enzyme adenosine deaminase (ADA) and has shown promise as an antileukemia agent. For the assessment of the extent to which systemically administered DCF crosses into the central nervous system (CNS), rhesus monkeys were given iv boluses of DCF. Simultaneous blood and cerebrospinal fluid (CSF) samples were assayed for DCF levels at times ranging from 10 minutes to 6 hours after the drug was given. Average peak CSF drug levels of 5.5 X 10(-8) M and 3 X 10(-7) M were reached 1 1/2 - 2 hours following injections of 0.25 and 1.0 mg DCF/kg, respectively. The ratio of peak CSF to simultaneous plasma levels was 1 to 10. Data obtained from a patient who had acute lymphocytic leukemia and who was given iv DCF were comparable. Drug levels achieved within the CSF following iv administration of 0.25 mg DCF/kg are similar to those previously demonstrated to inhibit ADA. These results may be important both for understanding DCF-related CNS toxicity and for designing combination chemotherapy with DCF.
Assuntos
Coformicina/líquido cefalorraquidiano , Leucemia Linfoide/líquido cefalorraquidiano , Ribonucleosídeos/líquido cefalorraquidiano , Animais , Barreira Hematoencefálica , Criança , Coformicina/administração & dosagem , Coformicina/análogos & derivados , Coformicina/sangue , Meia-Vida , Humanos , Injeções Intravenosas , Cinética , Macaca mulatta , Masculino , PentostatinaRESUMO
An analysis of the clinical outcomes in 66 patients with hairy cell leukemia treated with pentostatin under the Special Exception mechanism of the Division of Cancer Treatment, National Cancer Institute, between 1983 and 1987 has revealed a favorable balance of risk and benefit. Hematologic parameters and performance status were improved in most patients treated outside the clinical trials mechanism. The treating physicians considered 37 patients (56%) to be complete responders and 15 patients (23%) to be partial responders. Four patients (6%) died while receiving pentostatin. Life-threatening leukopenia (wbc count, less than 1,000/mm3) was reported in 24% of patients, and severe or life-threatening infection occurred in 11%. The experience gained with these patients supplements the information presently being collected from the controlled clinical trials and supports the development of a group C treatment protocol.
Assuntos
Antineoplásicos/uso terapêutico , Coformicina/uso terapêutico , Leucemia de Células Pilosas/tratamento farmacológico , Ribonucleosídeos/uso terapêutico , Adulto , Idoso , Idoso de 80 Anos ou mais , Antineoplásicos/efeitos adversos , Coformicina/efeitos adversos , Coformicina/análogos & derivados , Feminino , Humanos , Leucemia de Células Pilosas/patologia , Masculino , Pessoa de Meia-Idade , PentostatinaRESUMO
Thirty-one patients with hairy-cell leukemia were treated with 2'-deoxycoformycin (DCF) in a National Cancer Institute of Canada multicenter trial. The DCF was administered in a cycle (4 mg/m2 iv weekly X 3), which was repeated every 8 weeks. Following a complete remission, consolidation was done with two further cycles of DCF. Of 28 patients evaluable for response, 25 obtained a complete remission; 3 had a partial response. To date there has been only one relapse; the median time with no therapy was 429.5 days (range 99-743 days). Toxicity was moderate and included nausea and vomiting, lethargy, and skin rash; with the first cycle of treatment, neutropenia and an increased incidence of fever or infection were also observed. We conclude that low-dose DCF is highly effective in treating hairy-cell leukemia.
Assuntos
Antineoplásicos/uso terapêutico , Coformicina/uso terapêutico , Leucemia de Células Pilosas/tratamento farmacológico , Ribonucleosídeos/uso terapêutico , Antineoplásicos/efeitos adversos , Ensaios Clínicos como Assunto , Coformicina/efeitos adversos , Coformicina/análogos & derivados , Avaliação de Medicamentos , Feminino , Humanos , Leucemia de Células Pilosas/patologia , Masculino , Pentostatina , Indução de RemissãoRESUMO
2'-Deoxycoformycin (dCF), a tight-binding inhibitor of adenosine deaminase, has recently been entered into clinical trials. Toxicity has included lymphopenia, seizures, coma, conjunctivitis, renal failure, and hemolysis. Mice treated with dCF on a variety of schedules exhibited massive hemolysis. Hemolysis was brief, lasting about 20 hours, and did not recur upon readministration of the drug unless readministration was delayed for at least 6 days after initial exposure, which suggests that a sensitive subpopulation of cells was selectively destroyed. Splenectomy failed to protect the animals from dCF-induced hemolysis. Administration of adenosine or 2'-deoxyadenosine without dCF did not cause hemolysis, and use of these two agents with dCF did not potentiate the observed hemolysis. ATP and dATP levels were measured in erythrocytes, and changes in levels of these nucleotides did not correspond with the development of hemolysis.
Assuntos
Coformicina/toxicidade , Hemoglobinúria/induzido quimicamente , Hemólise/efeitos dos fármacos , Ribonucleosídeos/toxicidade , Animais , Coformicina/análogos & derivados , Relação Dose-Resposta a Droga , Avaliação de Medicamentos , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Feminino , Hematócrito , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Nucleotídeos/análise , Pentostatina , EsplenectomiaRESUMO
Adenosine (Ado) and Ado analogues produce multiple hemodynamic effects including coronary vasodilation, bradycardia, alterations in left ventricular contractility, and peripheral vasodilation or vasoconstriction depending on the vascular bed. The intact anesthetized Sprague-Dawley rat was examined in relation to electrocardiogram and blood pressure alterations induced by a series of potentially useful antineoplastic agents that are purine or pyrimidine analogues as part of a preclinical evaluation of these agents. The drugs tested were arabinosyladenine and its 5'-monophosphate derivative arabinosyladenine-5'-monophosphate (ara-AMP), the 2-fluoro derivative of ara-AMP, the pyrazolo[4,3-d]pyrimidines (formycin and formycin B), 8-azaadenosine, 6-methylmercaptopurine riboside, tricyclic nucleoside-5'-monophosphate, 5-fluorouracil, arabinosylcytosine, and 3-deazauridine. Those Ado analogues subject to deamination by adenosine deaminase (ADA) were also studied in the intact Sprague-Dawley rat after pretreatment with the ADA inhibitor 2'-deoxycoformycin. The results indicate that these agents have significant hemodynamic effects and should alert clinicians to potential adverse reactions when infusing these drugs.
Assuntos
Antineoplásicos/toxicidade , Hemodinâmica/efeitos dos fármacos , Hipotensão/induzido quimicamente , Adenosina/análogos & derivados , Adenosina/toxicidade , Inibidores de Adenosina Desaminase , Animais , Coformicina/análogos & derivados , Coformicina/farmacologia , Relação Dose-Resposta a Droga , Eletrocardiografia , Feminino , Formicinas/toxicidade , Pentostatina , Ratos , Ratos Endogâmicos , Fosfato de Vidarabina/análogos & derivados , Fosfato de Vidarabina/toxicidadeRESUMO
2'-Deoxycoformycin (DCF) is a potent inhibitor of adenosine deaminase (ADA) and a potential antineoplastic and immunosuppressive agent. In this study the kinetics of ADA expression was assessed by immunomorphologic and enzymatic methods in tissues of ACI rats given injections of DCF. The rats received a daily ip injection of 10 mg DCF/kg for 3 consecutive days. This treatment destroyed cortical thymocytes, whereas lymphocytes of the thymic medulla were mainly preserved. In control phosphate-buffered saline-injected rats, cortical thymocytes were not affected morphologically and displayed strong ADA staining. It was found unexpectedly that injections of DCF produced activation and, possibly, differentiation of B-cells in the mesenteric lymph nodes and spleen. These activated B-lymphocytes and plasma cells stained strongly for ADA. Transient changes in patterns of ADA expression were also observed in endothelial cells of blood vessels and liver Kupffer's cells, but these changes were not accompanied by degeneration of the cells. The treatment with DCF did not result in any permanent abnormalities in the rat tissues.
Assuntos
Adenosina Desaminase/análise , Coformicina/farmacologia , Nucleosídeo Desaminases/análise , Ribonucleosídeos/farmacologia , Timo/enzimologia , Inibidores de Adenosina Desaminase , Animais , Movimento Celular/efeitos dos fármacos , Coformicina/análogos & derivados , Duodeno/enzimologia , Feminino , Histocitoquímica , Técnicas Imunoenzimáticas , Linfonodos/enzimologia , Linfócitos/enzimologia , Masculino , Neutrófilos/enzimologia , Pentostatina , Plasmócitos/enzimologia , Ratos , Ratos Endogâmicos ACI , Baço/enzimologia , Timo/efeitos dos fármacos , Fatores de TempoRESUMO
2'-Deoxycoformycin (2'-dCF), a tight-binding inhibitor of adenosine deaminase, was administered to 26 pediatric patients with acute lymphoblastic leukemia in a Phase I study. Doses ranged from 0.25 to 1.0 mg/kg given i.v. for 3 consecutive days. Common toxicity included nausea, vomiting, diarrhea, hepatocellular enzyme elevations, and conjunctivitis. Lymphopenia occurred in all patients. The most serious adverse effects were acute tubular necrosis and central nervous system toxicity, which appeared to be dose related. In addition, two patients given the 0.75-mg/kg dose developed severe hepatic toxicity, although this could not be ascribed definitively to 2'-dCF. Antitumor activity was observed in eight patients, two of whom experienced a complete remission. Inhibition of lymphoblast adenosine deaminase activity was noted in the majority of cases and was observed at all doses. Antileukemic activity occurred at doses of 2'-dCF which were not associated with limiting toxicities. These results suggest that 2'-dCF is active against acute lymphoblastic leukemia and that a starting dose of 0.5 mg/kg/day be utilized in Phase II studies.
Assuntos
Coformicina/uso terapêutico , Leucemia Linfoide/tratamento farmacológico , Ribonucleosídeos/uso terapêutico , Inibidores de Adenosina Desaminase , Adolescente , Adulto , Criança , Pré-Escolar , Coformicina/efeitos adversos , Coformicina/análogos & derivados , Esquema de Medicação , Avaliação de Medicamentos , Feminino , Humanos , Fígado/efeitos dos fármacos , Linfopenia/induzido quimicamente , Masculino , Náusea/induzido quimicamente , Pentostatina , Prognóstico , Vômito/induzido quimicamenteRESUMO
2'-Deoxycoformycin (dCF), a potent inhibitor of adenosine deaminase, has recently undergone Phase I clinical trials and has been found to be therapeutically active in acute lymphoblastic leukemia. In this report, levels of dCF in plasma, plasma concentrations of adenosine and deoxyadenosine, and urine levels of deoxyadenosine were measured in leukemic patients undergoing treatment with dCF during a Phase I clinical trial. dCF was administered i.v. at a dose of 0.25 to 1.0 mg/kg (7.5 to 30 mg/sq m) for 3 consecutive days. Plasma drug levels of 2 to 6 microM were observed following the third dose of dCF, and drug accumulation occurred only at the 1-mg/kg dosage. In this limited series of patients, the plasma concentrations of adenosine and deoxyadenosine and the urine concentrations of deoxyadenosine did not show an obvious correlation with dCF dose, therapeutic response, or toxicity.
Assuntos
Adenosina/sangue , Coformicina/sangue , Desoxiadenosinas/sangue , Leucemia Linfoide/sangue , Ribonucleosídeos/sangue , Adolescente , Adulto , Criança , Pré-Escolar , Coformicina/análogos & derivados , Coformicina/uso terapêutico , Coformicina/urina , Desoxiadenosinas/urina , Avaliação de Medicamentos , Feminino , Humanos , Leucemia Linfoide/tratamento farmacológico , Masculino , Pentostatina , Fatores de TempoRESUMO
The antibiotic 2'-deoxycoformycin, a potent inhibitor of adenosine deaminase, has potential as a chemotherapeutic agent. Injection of 2'-deoxycoformycin i.v. (0.2 mg/kg) to mice bearing ascites L1210 leukemia cells completely inhibits adenosine deaminase in both erythrocytes and L1210 cells. The recovery of the enzymic activity is markedly different in the two tissues. The recovery is very slow in erythrocytes (13% in 48 hr), whereas 80% recovery occurs during the same time interval in L1210 cells. This marked difference in the recovery of the enzyme in different tissues may play a role in the pharmacological and chemotherapeutic behavior of this drug.
Assuntos
Inibidores de Adenosina Desaminase , Coformicina , Eritrócitos/efeitos dos fármacos , Leucemia L1210/tratamento farmacológico , Nucleosídeo Desaminases/antagonistas & inibidores , Ribonucleosídeos , Adenosina Desaminase/sangue , Animais , Coformicina/análogos & derivados , Coformicina/farmacologia , Eritrócitos/enzimologia , Leucemia L1210/enzimologia , Camundongos , Ribonucleosídeos/análogos & derivados , Fatores de TempoRESUMO
It has been suggested that, by inhibiting the adenosine deaminase (ADA)-mediated catabolism of 9-beta-D-arabinofuranosyladenine (ara-A), 2'-deoxycoformycin (DCF) would increase the half-life (t1/2) of ara-A, a compound with known antileukemic activity. To test this hypothesis, we collected serial plasma samples from five patients with refractory acute lymphoblastic leukemia who participated in a Phase I trial of i.v. DCF 915 mg/sq m) in combination with i.v. single-dose ara-A (120-250 mg/sq m). In four of these patients, of whom three were known to have achieved greater than 98% ADA inhibition, a mean ara-A t1/2 of 227 min was achieved. Extrapolated peak levels (i.e., following infusion of ara-A) ranged from 1.5 to 7.4 micrograms/ml (mean, 4.2 micrograms/ml). Elimination of drug appeared to follow a single-compartment model. In two patients who received ara-A without prior DCF and in a third patient who had significant residual ADA activity despite DCF, ara-A was unmeasurable within 5 min of the end of infusion. These data confirm that the kinetics of ara-A catabolism can be altered by inhibition of ADA and suggest that more than one dose of DCF may be necessary for complete inhibition of the enzyme and optimal pharmacological modulation of ara-A.
Assuntos
Coformicina/farmacologia , Leucemia Linfoide/tratamento farmacológico , Ribonucleosídeos/farmacologia , Vidarabina/metabolismo , Adolescente , Adulto , Arabinonucleosídeos/metabolismo , Pré-Escolar , Coformicina/análogos & derivados , Coformicina/uso terapêutico , Avaliação de Medicamentos , Quimioterapia Combinada , Feminino , Meia-Vida , Humanos , Hipoxantinas/metabolismo , Cinética , Leucemia Linfoide/metabolismo , Masculino , Pentostatina , Vidarabina/uso terapêuticoRESUMO
9-beta-D-Arabinofuranosyladenine (ara-A) inactivates isolated S-adenosyl-L-homocysteine (AdoHcy) hydrolase (EC 3.3.1.1) as well as AdoHcy hydrolase in intact cells. Whereas the inactivation in cell-free systems is an irreversible process, the AdoHcy hydrolase activity in rat hepatocytes exposed to ara-A gradually recovered upon prolonged incubation of the cells in a medium devoid of ara-A. This process, tentatively termed reactivation of the enzyme, was nearly totally dependent on a high level of adenosine deaminase in the extracellular medium, which induced a decrease in intracellular content of adenosine as well as ara-A. Reactivation of intracellular enzyme was inhibited by adenosine deaminase inhibitors [2'-deoxycoformycin and erythro-9-(2-hydroxy-3-nonyl)adenine] and the synthetic substrate for AdoHcy hydrolase, 3-deazaadenosine. An inhibitor of protein synthesis (cycloheximide) was without effect. Homocysteine, which protected the intracellular AdoHcy hydrolase against inactivation by ara-A, induced no reactivation of the enzyme. The half-life of the intracellular ara-A-AdoHcy hydrolase complex was about 90 min and was not affected by adenosine deaminase, 3-deazaadenosine, or homocysteine added to the cell suspension. However, the rate of elimination of the complex in the hepatocytes exceeded the rate of reactivation of AdoHcy hydrolase. Thus, the elimination process accounted for the reactivation, but not correlation between these two processes was observed. Reactivation of intracellular AdoHcy hydrolase caused a pronounced fall in cellular content of AdoHcy. The possibility that reduced cellular level of AdoHcy induced the reactivation of AdoHcy hydrolase seemed unlikely. This statement was based on the observation that reactivation was observed also under conditions of high concentrations of AdoHcy (obtained by the addition of homocysteine to the cell suspension). Reactivation of AdoHcy hydrolase with a concomitant decrease in cellular level of AdoHcy could also be demonstrated with mouse plasmacytoma (MPC-11) cells and mouse fibroblasts (L-929) exposed to ara-A, but the reactivation process was far less pronounced than with hepatocytes.
Assuntos
Hidrolases/metabolismo , Fígado/enzimologia , Vidarabina/farmacologia , Adenosina Desaminase/farmacologia , Inibidores de Adenosina Desaminase , Adenosil-Homocisteinase , Animais , Linhagem Celular , Células Cultivadas , Coformicina/análogos & derivados , Coformicina/farmacologia , Ativação Enzimática/efeitos dos fármacos , Fibroblastos/enzimologia , Homocisteína/farmacologia , Inosina/farmacologia , Camundongos , Pentostatina , Plasmocitoma/enzimologia , RatosRESUMO
There is a progressive loss of human T-lymphocyte viability upon incubation with deoxycoformycin, an adenosine deaminase inhibitor, and low concentrations of deoxyadenosine (drug concentration that reduced cell count at 48 hr after initiation to 50% of value for untreated control culture, less than 1 microM). The loss of viability was evidenced by vital staining with fluorescein diacetate and by changes in forward single light scatter measured by flow cytometry. This loss of lymphocyte viability is detectable 18 to 20 hr after the addition of deoxyadenosine and is earlier than has been reported by other investigators using trypan blue as the vital stain. Alkaline elution studies show that the incubation of T-lymphocytes with the combinations of deoxycoformycin and deoxyadenosine gives rise to DNA single-strand breaks. These DNA strand breaks are dose and time dependent and are readily detected 4 hr after the addition of deoxyadenosine. These DNA lesions are not observed with deoxycoformycin or deoxyadenosine alone. Incubations of T-lymphocytes with deoxycoformycin and deoxyadenosine (1 and 5 microM) for 7 hr result in DNA strand breaks with a frequency of 145 and 280 rad equivalents, respectively. Preliminary studies indicate that the ability of lymphocytes to repair this damage is dependent upon deoxyadenosine concentration and exposure time. The relationship of these DNA lesions to loss of lymphocyte viability in the presence of deoxycoformycin and deoxyadenosine remains to be established.
Assuntos
Antineoplásicos/toxicidade , Coformicina/toxicidade , DNA/genética , Desoxiadenosinas/toxicidade , Ribonucleosídeos/toxicidade , Linfócitos T/fisiologia , Células Cultivadas , Coformicina/análogos & derivados , DNA de Cadeia Simples/genética , Interações Medicamentosas , Humanos , Cinética , Pentostatina , Linfócitos T/efeitos dos fármacosRESUMO
The effect of the adenosine deaminase inhibitor, 2'-deoxycoformycin, on cellular nucleotides during therapy with continuous infusion of 9-beta-D-arabinofuranosyladenine (ara-A) has been investigated. In three courses of treatment using increasing doses, the active 5'-triphosphate of ara-A, 9-beta-D-arabinofuranosyladenine 5'-triphosphate (ara-ATP) accumulated in leukemic cells and erythrocytes from a patient treated for acute lymphocytic leukemia in proportion to the dose of ara-A. The cellular ara-ATP concentration increased more than 5-fold after the injection of a single, nontoxic, but pharmacologically active dose of 2'-deoxycoformycin 24 hr after initiation of ara-A infusion. However, this response was associated with a concomitant increase in the cellular deoxyadenosine triphosphate concentrations to levels equal to or greater than those of ara-ATP throughout the three treatment courses studied. Consistent with previous results using cell-free systems, it was demonstrated that a competitive relationship exists between deoxyadenosine triphosphate and ara-ATP for the inhibition of DNA synthesis in cultured human lymphoblastoid cells and that the ratio of the cellular concentrations of these nucleotides could predict the extent of inhibition of DNA synthesis. Application of this rationale to the nucleotides in the leukemic cells of the patient suggested that administration of 2'-deoxycoformycin may create a cellular biochemical milieu that could be antagonistic to the inhibition of DNA synthesis by ara-ATP.