RESUMO
Our laboratory has synthesized and evaluated the anticancer activity of a number of sulfonylhydrazine DNA modifying agents. As a class, these compounds possess broad spectrum antitumor activity, demonstrating significant activity against a variety of experimental murine tumors, including the P388 and L1210 leukemias, B16 melanoma, M109 lung carcinoma, and M5076 reticulum cell sarcoma, as well as against the human LX-1 lung carcinoma xenograft. The current report describes the activity of a more recently synthesized member of this class, 1,2-bis(methylsulfonyl)-1-(2-chloroethyl)-2-(methylamino)carbonylhydrazine (101M). 101M was active in mice against the i.p. implanted L1210 leukemia over a wide range of doses and produced long-term survivors when administered as a single i.p. bolus of 10, 20, 40, 60, or 80 mg/kg, demonstrating a wider margin of safety than the nitrosourea, 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). Curative therapy was achieved with doses of 101M that did not produce depression of the bone marrow. 101M was also highly effective against the L1210 leukemia when administered by the oral route. The ability of 101M to penetrate the blood-brain barrier and eradicate leukemia cells in the brain was remarkable (>6 log kill). This agent was also curative against L1210 variants resistant to cyclophosphamide, BCNU, or melphalan. Mice implanted with the murine C26 colon carcinoma were also cured by two injections of 10 or 20 mg/kg of 101M. Administration of 101M by two different well-tolerated regimens caused complete regression of established human glioblastoma U251 xenografts in 100% of treated mice, and significant responses were also obtained with 101M against advanced murine M109 lung carcinomas in mice. The broad spectrum of anticancer activity of the sulfonylhydrazine prodrug 101M coupled with the wide range of therapeutic safety exhibited by this agent, makes 101M particularly attractive for further development and clinical evaluation.
Assuntos
Antineoplásicos/farmacologia , Hidrazinas/farmacologia , Pró-Fármacos/farmacologia , Animais , Carmustina/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Humanos , Leucemia L1210/tratamento farmacológico , Melanoma Experimental/tratamento farmacológico , Camundongos , Camundongos Endogâmicos BALB C , Compostos Nitrosos/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Two major classes of plasma membrane proteins that actively extrude a wide range of structurally diverse hydrophobic amphipathic antineoplastic agents from cells, with different mechanisms of action, lead to multidrug resistance. To study the importance of these ATP-binding cassette transporters to the toxicity of cancer chemotherapy agents, we have used mice genetically deficient in both the mdr1a and mdr1b genes [mdr1a/1b(-/-) mice], the mrp1 gene [mrp1(-/-) mice], and the combined genes mdr1a/1b and mrp1 [mdr1a/1b(-/-), mrp1(-/-) mice] and embryonic fibroblasts derived from wild-type mice and from the three gene knockout animals. The consequences of export pump deficiencies were evaluated primarily using vincristine and etoposide. Mice deficient in the three genes, mdr1a/1b and mrp1, exhibited a 128-fold increase in toxicity to vincristine and a 3-5-fold increase in toxicity to etoposide; increased toxicity to embryonic fibroblast cells from triple knockout mice also occurred with vincristine and etoposide. Vincristine, which normally does not express toxicity to the bone marrow and to the gastrointestinal mucosa when used at therapeutic doses, caused extensive damage to these tissues in mdr1a/1b(-/-), mrp1(-/-) mice. The findings indicate that the P-glycoprotein and mrpl are compensatory transporters for vincristine and etoposide in the bone marrow and the gastrointestinal mucosa and emphasize the potential for increased toxicities by the combined inhibition of these efflux pumps.
Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Antineoplásicos Fitogênicos/toxicidade , Resistência a Múltiplos Medicamentos/genética , Etoposídeo/toxicidade , Genes MDR/genética , Vincristina/toxicidade , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Animais , Antineoplásicos Fitogênicos/farmacocinética , Western Blotting , Cruzamentos Genéticos , Relação Dose-Resposta a Droga , Etoposídeo/farmacocinética , Feminino , Masculino , Camundongos , Camundongos Knockout , Proteínas Associadas à Resistência a Múltiplos Medicamentos , Fenótipo , Vincristina/farmacocinéticaRESUMO
The mrp (multidrug resistance protein) gene has been associated with the multidrug resistance of cancer cells in vitro and in vivo. To gain information on its physiological role, embryonic stem cells were used to generate mice homozygous for a disruption of the mrp gene, resulting in complete abrogation of mrp expression. No physiological abnormalities were observed, at least up to 4 months of age. Viability, fertility, and a range of histological, hematological, and serum-chemical parameters were similar in mrp(+/+) and mrp(-/-) mice. mrp(-/-) mice displayed an increased sensitivity to etoposide phosphate (2-fold) accompanied by greater bone marrow toxicity, whereas the acute toxicity of sodium arsenite was equivalent in mrp(+/+) and mrp(-/-) mice. Tissue levels of glutathione (GSH) were elevated in breast, lung, heart, kidney, muscle, colon, testes, bone marrow cells, blood mononuclear leukocytes, and blood erythrocytes of mrp(-/-) mice and were unchanged in organs known to express little if any mrp, such as the liver and small intestine. The increase in GSH was not due to an increase in the activity of gamma-glutamylcysteine synthetase, the rate-limiting enzyme for GSH synthesis. The findings demonstrate that mrp is dispensable for development and growth but exerts a role in drug detoxification and GSH metabolism.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Antineoplásicos/toxicidade , Resistência a Múltiplos Medicamentos/genética , Etoposídeo/análogos & derivados , Etoposídeo/toxicidade , Glutationa/metabolismo , Compostos Organofosforados/toxicidade , Animais , Arsenitos/toxicidade , Medula Óssea/patologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Feminino , Glutamato-Cisteína Ligase/metabolismo , Masculino , Camundongos , Camundongos Knockout , Proteínas Associadas à Resistência a Múltiplos Medicamentos , Especificidade de Órgãos , Compostos de Sódio/toxicidade , Células-TroncoRESUMO
In preclinical investigations performed in mice, 2-amino-9-beta-D-ribofuranosyl purine-6-sulfinamide (sulfinosine), a novel derivative of 6-thioguanosine (6TGR), was active against six solid tumors and four strains of experimental leukemia. Sulfinosine penetrated the central nervous system more readily than did 6TGR and, when given repeatedly, was much more effective in the treatment of L1210 leukemia, being curative for some mice. Other findings of major interest to us were the different dosing characteristics of sulfinosine and 6TGR, the divergent efficiencies of the two drugs in generating cellular resistance, and the activity of sulfinosine against experimental leukemias refractory to 6TGR and other experimental or clinically used chemotherapeutic agents. The chemotherapeutic characterization of sulfinosine that evolved from these studies suggests that this agent may have unique properties that deserve clinical consideration. Both the dosing characteristics of the drug and its pronounced activity against thiopurine-resistant experimental leukemia favor the possibility that sulfinosine could be used to advantage in the treatment of human leukemia unresponsive to 6-mercaptopurine or 6-thioguanine.
Assuntos
Antineoplásicos/uso terapêutico , Neoplasias Experimentais/tratamento farmacológico , Nucleosídeos de Purina/uso terapêutico , Animais , Relação Dose-Resposta a Droga , Resistência a Medicamentos , Feminino , Guanosina/análogos & derivados , Guanosina/uso terapêutico , Leucemia L1210/tratamento farmacológico , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Tionucleosídeos/uso terapêuticoRESUMO
Site-selective cyclic AMP (cAMP) analogues inhibit growth and induce changes in morphology in a spectrum of human cancer cell lines (D. Katsaros et al., FEBS Lett., 223:97, 1987). The cellular events underlying such effects of cAMP analogues include differential regulation of type I versus type II cAMP-dependent protein kinase isozymes (S. Ally et al., Proc. Natl. Acad. Sci. USA, 85: 6319, 1988). Infusion (i.p.) of 8-Cl-cAMP, the most potent site-selective cAMP analogue, for 7 days produced regression of LX-1 lung carcinoma in athymic mice in a dose-dependent manner. The tumor regression correlated with the changing levels of cAMP receptor proteins, RI alpha and RII beta, the regulatory subunits of cAMP-dependent protein kinase type I and type II, respectively. By photoaffinity labeling with 8-N3-[32P]cAMP and immunoblotting with a monospecific anti-RII antibody, RI alpha (Mr 49,000) and RII beta (Mr 51,000) were identified in the untreated control tumors. 8-Cl-cAMP treatment induced a rapid increase of both RI alpha and RII beta in tumor cytosols and translocation (within 1 h) of only RII beta from the cytosol to the nucleus. RII beta in both cytosols and nuclei remained elevated during 8-Cl-cAMP treatment, whereas RI alpha in the cytosols gradually decreased with time of treatment after its initial transient increase. Northern blot analyses demonstrated that the RII beta mRNA level increased within 6 h of 8-Cl-cAMP treatment and remained elevated during treatment, whereas the RI alpha mRNA level decreased to below that of the untreated control tumor level after its transient increase during 1-6 h of treatment. 8-Cl-cAMP treatment also caused a sharp decrease in both N-ras and c-myc mRNA levels. These results suggest that the fundamental basis for the antineoplastic activity of 8-Cl-cAMP may reside in the restoration of normal gene regulation in neoplasms in which cAMP receptor proteins play a role.
Assuntos
Carcinoma/genética , Divisão Celular/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Neoplasias Pulmonares/genética , Proteínas Quinases/metabolismo , Receptores de AMP Cíclico/metabolismo , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , 8-Bromo Monofosfato de Adenosina Cíclica/uso terapêutico , Animais , Northern Blotting , Western Blotting , Carcinoma/tratamento farmacológico , Carcinoma/metabolismo , Núcleo Celular/metabolismo , Genes ras , Humanos , Isoenzimas/metabolismo , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/metabolismo , Camundongos , Camundongos Nus , Transplante de Neoplasias , Oncogenes , RNA Mensageiro/metabolismo , RNA Neoplásico/metabolismoRESUMO
Embryonic chick muscle cells in the infused (myoblast) and fused (myotube) states differentially incorporate [3H]palmitic acid into their cellular lipids. Myoblasts incorporated [3H]palmitic acid 6--7-fold greater than myotubes in the triglyceride fraction. The endogenous triglyceride levels, however, were similar for the two developmental stages indicating increased triglyceride turnover in the myoblast compared to that of the myotube. Myotubes showed an increased incorporation of [3H]palmitic acid into sphingomyelin compared to myoblasts. Myoblasts and myotubes had a similar phospholipid composition. The data indicate that fusion is characterized by a decrease in triglyceride turnover possibly reflecting a shift from oxidative to glycolytic metabolism to coincide with the fusion process.
Assuntos
Metabolismo dos Lipídeos , Músculos/metabolismo , Animais , Células Cultivadas , Embrião de Galinha , Ácidos Graxos/análise , Músculos/citologia , Músculos/embriologia , Ácidos Palmíticos/metabolismo , Fosfolipídeos/metabolismo , Triglicerídeos/metabolismoRESUMO
Glycosylation of 1,2,4 lambda 3-diazaphosphole (4) under Lewis acid catalyzed conditions gave 1-alpha-D-ribofuranosyl-1,2,4 lambda 3-diazaphosphole (5) as the only product. Ethyl 1,2,4 lambda 3-diazaphosphole-3-carboxylate (10) was synthesized by the cyclocondensation of ethyl (chlorophosphinidene)(trimethylsilyl)acetate (8) with (trimethylsilyl)diazomethane and subsequent desilylation with tetra-n-butylammonium fluoride. Reaction of 10 with methanolic ammonia at 80 degrees C gave 1,2,4 lambda 3-diazaphosphole-3-carboxamide. Glycosylation of 10 using trimethylsilyl triflate catalyst followed by ammonlysis gave the ribavirin (1) analogue 1-beta-D-ribofuranosyl-1,2,4 lambda 3-diazaphosphole-3-carboxamide (11). Acetylation of 11 and subsequent treatment with phosphorus pentasulfide gave 2',3',5'-tri-O-acetyl-1-beta-D-ribofuranosyl-1,2,4 lambda 3-diazaphosphole-3- thiocarboxamide (13). Deprotection with methanolic ammonia gave 1-beta-D-ribofuranosyl-1,2,4 lambda 3-diazaphosphole-3-thiocarboxamide (14). Compound 14 gave a 25% increase in life span (ILS) against L1210 in female BDF1 mice. The anomeric configuration and site of glycosylation of 5 and 13 were established by single-crystal X-ray crystallography.
Assuntos
Antineoplásicos/síntese química , Azidas/síntese química , Nucleosídeos/síntese química , Compostos Organofosforados/síntese química , Animais , Antineoplásicos/efeitos adversos , Antineoplásicos/uso terapêutico , Azidas/efeitos adversos , Azidas/uso terapêutico , Fenômenos Químicos , Química , IMP Desidrogenase/antagonistas & inibidores , Camundongos , Modelos Moleculares , Neoplasias Experimentais/tratamento farmacológico , Compostos Organofosforados/efeitos adversos , Compostos Organofosforados/uso terapêutico , Relação Estrutura-Atividade , Análise de Sobrevida , Difração de Raios XRESUMO
7-Deaza (pyrrolo[2,3-d]pyrimidine) and 3-deaza (imidazo[4,5-c]pyridine) congeners of sulfenosine (5a and 9), sulfinosine (6a and 10), and sulfonosine (7a) have been prepared and evaluated for their antileukemic activity in mice. Amination of 2-amino-7-beta-D-ribofuranosylpyrrolo[2,3-d]pyrimidine-4(3H)-th ion e (4a) and its 2'-deoxy analogue (4c) with a chloramine solution gave the corresponding 4-sulfenamides (5a and 5c, respectively), which on selective oxidation with m-chloroperoxybenzoic acid (MCPBA) gave the respective diastereomeric 2-amino-7-beta-D-ribofuranosyl-pyrrolo[2,3-d]pyrimidine-4-sulfinamide (7-deazasulfinosine, 6a) and its 2'-deoxy derivative (6c). A similar amination of 7-(2-deoxy-beta-D-erythro-pentofuranosyl)pyrrolo[2,3-d]pyrimidine-4(3H)- thione (4b) gave the corresponding 4-sulfenamide derivative (5b). Oxidation of 5b with 1 molar equiv of MCPBA furnished (R,S)-7-(2-deoxy-beta-D-erythro-pentofuranosyl)pyrrolo[2,3-d]pyrimidine- 4- sulfinamide (6b), whereas use of excess of MCPBA afforded the corresponding sulfonamide derivative (7b). Treatment of 3-deaza-6-thioguanosine (8) with a chloramine solution gave 3-deazasulfenosine (6-amino-1-beta-D- ribofuranosylimidazo[4,5-c]pyridine-4-sulfenamide, 9). Controlled oxidation of 9 with MCPBA afforded 3-deazasulfinosine (10). As gauged by increases in the mean postinoculation life spans of L1210 inoculated mice, none of these nucleosides exhibited biologically significant activity (T/C greater than or equal to 125). Even so, antileukemic activity appeared to be influenced, albeit not uniformly, by structural modifications in the base and carbohydrate moieties of sulfenosine and sulfinosine. Thus, while several of the compounds were lacking in cytotoxic activity, eight others (4c, 5a, 5c, 6a, 6b, 7b, 9, and 10) were estimated to have reduced body burdens of viable L1210 cells by 16-77%.
Assuntos
Antineoplásicos/síntese química , Nucleosídeos de Purina/síntese química , Nucleosídeos de Pirimidina/síntese química , Ribonucleosídeos/uso terapêutico , Animais , Fenômenos Químicos , Química , Feminino , Leucemia L1210/tratamento farmacológico , Camundongos , Nucleosídeos de Purina/uso terapêutico , Nucleosídeos de Pirimidina/farmacologia , Relação Estrutura-Atividade , Sulfetos/uso terapêutico , Sulfonamidas/uso terapêutico , Sulfóxidos/uso terapêuticoRESUMO
A number of 6-sulfenamide, 6-sulfinamide, and 6-sulfonamide derivatives of 2-aminopurine and certain related purine ribonucleosides have been synthesized and evaluated for antileukemic activity in mice. Amination of 6-mercaptopurine ribonucleoside (7a) and 6-thioguanosine (7b) with chloramine solution gave 9-beta-D-ribofuranosylpurine-6-sulfenamide (8a) and 2-amino-9-beta-D-ribofuranosylpurine-6-sulfenamide (sulfenosine, 8b), respectively. Selective oxidation of 8a and 8b with 3-chloroperoxybenzoic acid (MCPBA) gave (R,S)-9-beta-D-ribofuranosylpurine-6-sulfinamide (9a) and (R,S)-2-amino-9-beta-D-ribofuranosylpurine-6-sulfinamide (sulfinosine, 9b), respectively. However, oxidation of 8a and 8b with excess of MCPBA gave 9-beta-D-ribofuranosylpurine-6-sulfonamide (10a) and 2-amino-9-beta-D-ribofuranosylpurine-6-sulfonamide (sulfonosine, 10b), respectively. Similarly, amination of 5'-deoxy-6-thioguanosine (7c) afforded the 6-sulfenamide derivative (8c), which on controlled oxidation gave (R,S)-2-amino-9-(5-deoxy-beta-D-ribofuranosyl)purine-6-sulfinamide (9c) and the corresponding 6-sulfonamide derivative (10c). Treatment of 6-thioguanine (12) with aqueous chloramine solution gave 2-amino-9H-purine-6-sulfenamide (13). Oxidation of 13 with 1 molar equiv of MCPBA afforded (R,S)-2-amino-9H-purine-6-sulfinamide (14), whereas the use of 4 molar equiv of MCPBA furnished 2-amino-9H-purine-6-sulfonamide (15). The resolution of R and S diastereomers of sulfinosine (9b) was accomplished by HPLC techniques. The structures of (R)-9b and 10b were assigned by single-crystal X-ray diffraction studies. (R)-9b exists in the crystal structure in four crystallographically independent conformations. Of the 18 compounds evaluated, 13 exhibited very significant anti-L1210 activity in mice. Sulfenosine (8b) at 22 mg/kg per day X 1 showed a T/C of 170, whereas sulfinosine (9b) at 173 mg/kg per day X 1 showed a T/C of 167 against L1210 leukemia. The 5'-deoxy analogue of sulfinosine (9c) at 104 mg/kg per day also showed a T/C of 172. A single treatment with 8b, 9b, and 9c reduced body burdens of viable L1210 cells by more than 99.8%.
Assuntos
2-Aminopurina/análogos & derivados , Adenina/análogos & derivados , Antineoplásicos , Nucleosídeos de Purina/uso terapêutico , Ribonucleosídeos/uso terapêutico , Sulfonamidas/uso terapêutico , 2-Aminopurina/síntese química , 2-Aminopurina/uso terapêutico , Animais , Fenômenos Químicos , Química , Feminino , Leucemia L1210/tratamento farmacológico , Camundongos , Conformação Molecular , Estrutura Molecular , Nucleosídeos de Purina/síntese química , Ribonucleosídeos/síntese química , Sulfonamidas/síntese química , Difração de Raios XRESUMO
The intracellular accumulation of cAMP inhibits the growth of transformed cells in vitro and in vivo, and exposure to various cAMP analogs produces similar results. The influence of such analogs on the growth of neoplastic cells in vivo is less well defined, and the relevance of these analogs for the phosphoinositide pathway has not been established. The present report details the inhibition of tumor growth that occurred when human mammary xenografts were treated with 8-Cl-cAMP, the subsequent rebound in tumor growth that occurred when treatment ceased, and the levels of diacylglycerol and membrane-associated protein kinase C activity that characterized tumors in different growth states. Tumor levels of diacylglycerol and particulate PKC activity appeared to be influenced not only by treatment but also by treatment withdrawal. Changes in these entities tended to coincide with tumor growth rate, being relatively suppressed during growth stasis and markedly elevated during periods of rapid growth. The data presented do not establish a causal relationship. Thus, the concomitant changes noted in tumor growth and tumor levels of either diacylglycerol and membrane-associated protein kinase C may only be coincidental. Alternatively, they may indicate that cAMP analogs inhibit tumor growth in vivo by modulating the phosphoinositide pathway.
Assuntos
8-Bromo Monofosfato de Adenosina Cíclica/análogos & derivados , Antineoplásicos/farmacologia , Diglicerídeos/análise , Glicerídeos/análise , Neoplasias Mamárias Experimentais/patologia , Proteína Quinase C/análise , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Adulto , Animais , Membrana Celular/enzimologia , Feminino , Humanos , Neoplasias Mamárias Experimentais/análise , Camundongos , Transplante de Neoplasias , Transplante HeterólogoRESUMO
Preclinical investigations in vivo revealed unexpected differences in the biological characteristics of 2-amino-9-beta-D-ribofuranosylpurine-6-sulfenamide (sulfenosine, 1) and 2-amino-9-beta-D-ribofuranosylpurine-6-sulfonamide (sulfonosine, 2), two novel but structurally related derivatives of 6-thioguanosine (6TGR). Strikingly, the addition of a fully oxidized sulfur atom at the 6 position of sulfenosine produced a purine derivative (sulfonosine) that was remarkably active against experimental leukemia resistant to treatment with either sulfenosine or 6TGR. This slight structural modification also appeared to influence solubility, scheduling capability, and oral activity as well as penetration of the central nervous system (CNS) and the onset of cellular resistance. These findings underscore the dramatic changes in biologic activity that can be produced by subtle modifications in molecular structure. We trust they may also contribute to the development of improved clinical therapy.
Assuntos
Antineoplásicos/uso terapêutico , Guanosina/análogos & derivados , Leucemia L1210/tratamento farmacológico , Nucleosídeos de Purina/uso terapêutico , Sulfonamidas/uso terapêutico , Tionucleosídeos/uso terapêutico , Animais , Barreira Hematoencefálica , Resistência a Medicamentos , Feminino , Guanosina/farmacocinética , Guanosina/uso terapêutico , Camundongos , Oxirredução , Relação Estrutura-Atividade , Tionucleosídeos/farmacocinéticaRESUMO
Growing interest in the MRP (multidrug resistance protein) gene stems from its importance in multidrug resistance to chemotherapy, its possible use in gene therapy, and its relationship with the glutathione system. The recent generation of mrp gene knockout models in vitro and in vivo is providing information on the mechanism of action and the physiological function(s) of mrp. The importance of mrp in protection of normal tissues from the toxicity of the anticancer agent etoposide has been established. A total block of mrp has been found to be compatible with life, suggesting that MRP inhibitors can be safely used for treating cancer patients. In some sub-classes of leukocytes, mrp contributes to the transport of leukotriene C4, an endogenous glutathione-S-conjugate. However, the baseline expression of mrp does not appear to contribute to the export of glutathione-S-conjugates of alkylating agents, and thus does not exert a protective role against their toxicity. Besides being capable of exporting certain glutathione-S-conjugates, mrp also catalyzes the co-transport of GSH and drug and, presumably, a presently unknown endogenous metabolite(s).
Assuntos
Resistencia a Medicamentos Antineoplásicos/fisiologia , Genes MDR , Animais , Antineoplásicos Fitogênicos/farmacologia , Transporte Biológico , Proteínas de Transporte/metabolismo , Etoposídeo/farmacologia , Glutationa/metabolismo , Humanos , Leucotrieno C4/metabolismo , Proteínas de Membrana Transportadoras , Metais Pesados/metabolismo , Camundongos , Camundongos Knockout , Canais de Potássio/metabolismoRESUMO
The basis for the antitumor activities of the exocyclic amino nucleosides 4-amino-(ARPP) and 4-methoxy-8-(D-ribofuranosylamino)pyrimido[5,4-d]pyrimidine (MRPP) was investigated. The primary target of these nucleosides appeared to be 5-phospho-alpha-D-ribofuranose-1-pyrophosphate (PRPP) synthetase. MRPP-5'-monophosphate was a competitive inhibitor (Ki = 40 microM) of the activation of this enzyme by the cofactor inorganic phosphate (K alpha = 2.2 mM). Consequently, ARPP and MRPP treatment of WI-L2 cultures rapidly inhibited both de novo pyrimidine and purine synthesis as well as the nucleotide salvage reactions dependent on PRPP, ARPP or MRPP treatment completely prevented [14C]bicarbonate incorporation into acid-soluble pyrimidine and purine nucleotides. The rate of salvage of [8-14C]hypoxanthine to form IMP was decreased by 85%. Treatment of cells with these agents caused a 50% reduction in the steady-state level of PRPP. When the capacity of the treated cells for sustained synthesis of PRPP was examined by adenine incorporation, the rate of adenine uptake was inhibited by greater than 50%. In vivo treatment of BDF1 mice with a single dose of ARPP (173 mg/kg) or MRPP (62 mg/kg) extended the mean life span of the mice, which had been inoculated intraperitoneally 1 day earlier with 1 x 10(6) L1210 murine leukemia cells, by 62 and 82% respectively. These studies indicate that MRPP and ARPP inhibit PRPP synthetase, and that PRPP synthetase may be a viable target in the development of certain antitumor agents.
Assuntos
Antineoplásicos/farmacologia , Fosfotransferases/antagonistas & inibidores , Nucleosídeos de Pirimidina/farmacologia , Ribose-Fosfato Pirofosfoquinase/antagonistas & inibidores , Adenosina Desaminase/farmacologia , Adenosina Quinase/farmacologia , Animais , Feminino , Camundongos , NAD/metabolismo , Nucleotídeos/biossíntese , Fosforribosil Pirofosfato/análise , Fosforribosil Pirofosfato/biossínteseRESUMO
Previous studies from our laboratories have shown that (a) Triapine() is a potent inhibitor of ribonucleotide reductase activity and (b) hydroxyurea-resistant L1210 leukemia cells are fully sensitive to Triapine. In an analogous manner, Triapine was similarly active against the wild-type and a hydroxyurea-resistant subline of the human KB nasopharyngeal carcinoma. Triapine was active in vivo against the L1210 leukemia over a broad range of dosages and was curative for some mice. This agent also caused pronounced inhibition of the growth of the murine M109 lung carcinoma and human A2780 ovarian carcinoma xenografts in mice. Optimum anticancer activity required twice daily dosing due to the duration of inhibition of DNA synthesis which lasted about 10 hr in L1210 cells treated with Triapine in vivo. DNA synthesis in normal mouse tissues (i.e. duodenum and bone marrow) uniformly recovered faster than that in L1210 leukemia cells, demonstrating a pharmacological basis for the therapeutic index of this agent. Triapine was more potent than hydroxyurea in inhibiting DNA synthesis in L1210 cells in vivo, and the effects of Triapine were more pronounced. In addition, the duration of the inhibition of DNA synthesis in leukemia cells from mice treated with Triapine was considerably longer than in those from animals treated with hydroxyurea. Combination of Triapine with various classes of agents that damage DNA (e.g. etoposide, cisplatin, doxorubicin, and 1-acetyl-1,2-bis(methylsulfonyl)-2-(2-chloroethyl)hydrazine) resulted in synergistic inhibition of the L1210 leukemia, producing long-term survivors of tumor-bearing mice treated with several dosage levels of the combinations, whereas no enhancement of survival was found when Triapine was combined with gemcitabine or cytosine arabinoside. The findings demonstrate the superiority of Triapine over hydroxyurea as an anticancer agent and further suggest that prevention by Triapine of repair of DNA lesions created by agents that damage DNA may result in efficacious drug combinations for the treatment of cancer.
Assuntos
Antineoplásicos/farmacologia , Inibidores Enzimáticos/farmacologia , Piridinas/farmacologia , Ribonucleotídeo Redutases/antagonistas & inibidores , Tiossemicarbazonas/farmacologia , Animais , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Barreira Hematoencefálica , Divisão Celular/efeitos dos fármacos , DNA/biossíntese , DNA/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos , Inibidores Enzimáticos/farmacocinética , Inibidores Enzimáticos/uso terapêutico , Humanos , Hidroxiureia/farmacologia , Células KB , Leucemia L1210/tratamento farmacológico , Camundongos , Transplante de Neoplasias , Neoplasias Experimentais/tratamento farmacológico , Piridinas/farmacocinética , Piridinas/uso terapêutico , Tiossemicarbazonas/farmacocinética , Tiossemicarbazonas/uso terapêutico , Células Tumorais CultivadasRESUMO
A workshop titled "Using Sentinel Species Data to Address the Potential Human Health Effects of Chemicals in the Environment," sponsored by the U.S. Army Center for Environmental Health Research, the National Center for Environmental Assessment of the EPA, and the Agency for Toxic Substances and Disease Registry, was held to consider the use of sentinel and surrogate animal species data for evaluating the potential human health effects of chemicals in the environment. The workshop took a broad view of the sentinel species concept, and included mammalian and nonmammalian species, companion animals, food animals, fish, amphibians, and other wildlife. Sentinel species data included observations of wild animals in field situations as well as experimental animal data. Workshop participants identified potential applications for sentinel species data derived from monitoring programs or serendipitous observations and explored the potential use of such information in human health hazard and risk assessments and for evaluating causes or mechanisms of effect. Although it is unlikely that sentinel species data will be used as the sole determinative factor in evaluating human health concerns, such data can be useful as for additional weight of evidence in a risk assessment, for providing early warning of situations requiring further study, or for monitoring the course of remedial activities. Attention was given to the factors impeding the application of sentinel species approaches and their acceptance in the scientific and regulatory communities. Workshop participants identified a number of critical research needs and opportunities for interagency collaboration that could help advance the use of sentinel species approaches.
Assuntos
Exposição Ambiental/efeitos adversos , Saúde Ambiental , Monitoramento Ambiental/métodos , Poluentes Ambientais/efeitos adversos , Vigilância de Evento Sentinela , Animais , Bioensaio , Humanos , Medição de Risco , Vigilância de Evento Sentinela/veterinária , Especificidade da Espécie , Estados UnidosRESUMO
The enzyme RR catalyzes the conversion of ribonucleoside diphosphates to their deoxyribonucleotide counterparts. RR is critical for the generation of the cytosine, adenine, and guanine deoxyribonucleotide 5'-triphosphate building blocks of DNA, which are present in cells as exceedingly small intracellular pools. Therefore, interference with the function of RR might well result in an agent with significant antineoplastic activity, particularly against rapidly proliferating tumor cells. HUr is the only inhibitor of RR in clinical usage; this agent, however, is a relatively poor inhibitor of the enzyme and has a short serum half-life. Consequently, HUr is a relatively weak anticancer agent. In an effort to develop a more potent inhibitor of RR with utility as an anticancer agent, we have synthesized 3-AP and demonstrated (a) potent inhibition of L1210 leukemia cells in vitro, (b) curative capacity for mice bearing the L1210 leukemia, (c) marked inhibition of RR, and (d) sensitivity of HUr-resistant cells to 3-AP. These findings collectively demonstrate the clinical potential of 3-AP as an antineoplastic agent.
Assuntos
Antineoplásicos/farmacologia , Inibidores Enzimáticos/farmacologia , Piridinas/farmacologia , Ribonucleotídeo Redutases/antagonistas & inibidores , Tiossemicarbazonas/farmacologia , Animais , Divisão Celular/efeitos dos fármacos , DNA de Neoplasias/biossíntese , Resistência a Medicamentos , Feminino , Hidroxiureia/farmacologia , Técnicas In Vitro , Ferro/metabolismo , Leucemia L1210/tratamento farmacológico , Leucemia L1210/metabolismo , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos DBA , Células Tumorais CultivadasRESUMO
The observation that cells often respond to carcinogens by nuclear enlargement has suggested that this property might be useful to develop a short-term screening test for such compounds. Previous methods for detecting nuclear size increases have used an image analyzer system to detect nuclear changes in individual cells. This paper details a more rapid method for obtaining nuclei by use of a stromalyzing procedure following by analysis of nuclear volumes, using a Coulter Counter Channelyzer. This new and simplified nuclear sizing method should facilitate the use of the assay as a possible carcinogenesis screen by permitting rapid and efficient testing of large numbers of compounds.
Assuntos
Carcinógenos , Núcleo Celular/efeitos dos fármacos , Contagem de Células , Cicloeximida/toxicidade , Ciclofosfamida/toxicidade , Técnicas Citológicas , Relação Dose-Resposta a Droga , Etídio/toxicidade , Células HeLa/efeitos dos fármacos , Humanos , Técnicas In Vitro , Mecloretamina/toxicidade , Metilnitronitrosoguanidina/toxicidade , Fatores de TempoRESUMO
A patient being treated for leukemia received lithium carbonate and ticarcillin for sepsis, and polyuria and severe hypernatremia developed. Although useful in neutropenic patients, the simultaneous use of these drugs may result in life-threatening hypernatremia.
Assuntos
Hipernatremia/induzido quimicamente , Lítio/efeitos adversos , Penicilinas/efeitos adversos , Ticarcilina/efeitos adversos , Idoso , Feminino , Humanos , Carbonato de LítioRESUMO
A case of avulsion of a unilateral axillofemoral Dacron bypass graft 3 weeks after insertion is reported. Surface measurements on healthy volunteers suggest that when the thoracolumbar spine is in full lateral flexion the length of the graft may increase by up to 21 per cent. Such movements should be restricted during the early postoperative period.