RESUMO
Herein we describe the discovery of IDX21437 35b, a novel RPd-aminoacid-based phosphoramidate prodrug of 2'-α-chloro-2'-ß-C-methyluridine monophosphate. Its corresponding triphosphate 6 is a potent inhibitor of the HCV NS5B RNA-dependent RNA polymerase (RdRp). Despite showing very weak activity in the in vitro Huh-7 cell based HCV replicon assay, 35b demonstrated high levels of active triphosphate 6 in mouse liver and human hepatocytes. A biochemical study revealed that the metabolism of 35b was mainly attributed to carboxyesterase 1 (CES1), an enzyme which is underexpressed in HCV Huh-7-derived replicon cells. Furthermore, due to its metabolic activation, 35b was efficiently processed in liver cells compared to other cell types, including human cardiomyocytes. The selected RP diastereoisomeric configuration of 35b was assigned by X-ray structural determination. 35b is currently in Phase II clinical trials for the treatment of HCV infection.
Assuntos
Antivirais/farmacologia , RNA Polimerases Dirigidas por DNA/antagonistas & inibidores , Descoberta de Drogas , Inibidores Enzimáticos/farmacologia , Hepacivirus/efeitos dos fármacos , Uridina Monofosfato/análogos & derivados , Uridina/farmacologia , Animais , Antivirais/síntese química , Antivirais/química , RNA Polimerases Dirigidas por DNA/metabolismo , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Hepacivirus/enzimologia , Hepatócitos/efeitos dos fármacos , Hepatócitos/virologia , Humanos , Fígado/efeitos dos fármacos , Fígado/virologia , Camundongos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Relação Estrutura-Atividade , Uridina/síntese química , Uridina/química , Uridina Monofosfato/síntese química , Uridina Monofosfato/química , Uridina Monofosfato/farmacologia , Proteínas não Estruturais Virais/antagonistas & inibidores , Proteínas não Estruturais Virais/metabolismoRESUMO
After decades of intensive research on adenosine-3',5'-cyclic monophosphate (cAMP)- and guanosine-3',5'-cyclic monophosphate (cGMP)-related second messenger systems, also the noncanonical congeners cyclic cytidine-3',5'-monophosphate (cCMP) and cyclic uridine-3',5'-monophosphate (cUMP) gained more and more interest. Until the late 1980s, only a small number of cCMP and cUMP analogs with sometimes undefined purities had been described. Moreover, most of these compounds had been rather synthesized as precursors of antitumor and antiviral nucleoside-5'-monophosphates and hence had not been tested for any second messenger activity. Along with the recurring interest in cCMP- and cUMP-related signaling in the early 2000s, it became evident that well-characterized small molecule analogs with reliable purities would serve as highly valuable tools for the evaluation of a putative second messenger role of cyclic pyrimidine nucleotides. Meanwhile, for this purpose new cCMP and cUMP derivatives have been developed, and already known analogs have been resynthesized and highly purified. This chapter summarizes early medicinal chemistry work on cCMP and cUMP and analogs thereof, followed by a description of recent synthetic developments and an outlook on potential future directions.
Assuntos
CMP Cíclico/síntese química , Nucleotídeos Cíclicos/síntese química , Pró-Fármacos/síntese química , Uridina Monofosfato/síntese química , Animais , Cristalização , CMP Cíclico/análogos & derivados , CMP Cíclico/metabolismo , CMP Cíclico/farmacologia , Humanos , Estrutura Molecular , Nucleotídeos Cíclicos/metabolismo , Nucleotídeos Cíclicos/farmacologia , Permeabilidade , Pró-Fármacos/metabolismo , Pró-Fármacos/farmacologia , Uridina Monofosfato/metabolismo , Uridina Monofosfato/farmacologiaRESUMO
Nucleoside-(5'âP) methylenebisphosphonodithioate analogues are bioisosteres of natural nucleotides. The potential therapeutic applications of these analogues are limited by their relative instability. With a view toward improving their chemical and metabolic stability as well as their affinity toward zinc ions, we developed a novel nucleotide scaffold, nucleoside-5'-tetrathiobisphosphonate. We synthesized P1-(uridine/adenosine-5')-methylenebisphosphonodithioate, 2 and 3, and P1,P2-di(uridine/adenosine-5')-methylenebisphosphonodithioate, 4 and 5. Using (1)H and (31)P NMR-monitored Zn(2+)/Mg(2+) titrations, we found that 5 coordinated Zn(2+) by both N7 nitrogen atoms and both dithiophosphonate moieties, whereas 3 coordinated Zn(2+) by an N7 nitrogen atom and Pß. Both 3 and 5 did not coordinate Mg(2+) ions. (31)P NMR-monitored kinetic studies showed that 3 was more stable at pD 1.5 than 5, with t(1/2) of 44 versus 9 h, respectively, and at pD 11 both showed no degradation for at least 2 weeks. However, 5 was more stable than 3 under an air-oxidizing atmosphere, with t1/2 of at least 3 days versus 14 h, respectively. Analogues 3 and 5 were highly stable to NPP1,3 and NTPDase1,2,3,8 hydrolysis (0-7%). However, they were found to be poor ectonucleotidase inhibitors. Although 3 and 5 did not prove to be effective inhibitors of zinc-containing NPP1/3, which is involved in the pathology of osteoarthritis and diabetes, they may be promising zinc chelators for the treatment of other health disorders involving an excess of zinc ions.
Assuntos
Monofosfato de Adenosina/análogos & derivados , Difosfonatos/química , Nucleosídeos/química , Nucleosídeos/síntese química , Compostos Organotiofosforados/química , Uridina Monofosfato/análogos & derivados , Monofosfato de Adenosina/síntese química , Monofosfato de Adenosina/química , Difosfonatos/síntese química , Estrutura Molecular , Compostos Organotiofosforados/síntese química , Uridina Monofosfato/síntese química , Uridina Monofosfato/químicaRESUMO
Prodrugs of therapeutic nucleoside monophosphates masked as phosphoramidate derivatives have become an increasingly important class of antiviral drugs in pharmaceutical research for delivering nucleotides in vitro and in vivo. Conventionally, phosphoramidate derivatives are prepared as a mixture of two diastereomers. We report a class of stable phosphoramidating reagents containing an amino acid ester and two phenolic groups, one unsubstituted and the other with electron-withdrawing substituents. The reagents can be isolated as single diastereomers and reacted with the 5'-hydroxyl group of nucleosides through selective nucleophilic displacement of the substituted phenol to prepare single diastereomer phosphoramidate products. This method has been used to prepare the HCV clinical candidate PSI-7977 in high yield and high diastereomeric purity.
Assuntos
Amidas/química , Antivirais/síntese química , Química Farmacêutica/métodos , RNA Polimerases Dirigidas por DNA/antagonistas & inibidores , Hepacivirus/efeitos dos fármacos , Hepatite C/tratamento farmacológico , Ácidos Fosfóricos/química , Uridina Monofosfato/análogos & derivados , Proteínas Virais/antagonistas & inibidores , Aminoácidos/química , Antivirais/farmacologia , Cromatografia Líquida de Alta Pressão , RNA Polimerases Dirigidas por DNA/metabolismo , Elétrons , Ésteres/química , Hepacivirus/enzimologia , Hepatite C/virologia , Humanos , Espectroscopia de Ressonância Magnética , Nucleosídeos/química , Nucleotídeos/química , Fenóis/química , Pró-Fármacos/síntese química , Pró-Fármacos/farmacologia , Sofosbuvir , Estereoisomerismo , Relação Estrutura-Atividade , Uridina Monofosfato/síntese química , Uridina Monofosfato/farmacologia , Proteínas Virais/metabolismoRESUMO
Irradiation of solutions at pH 7 to pH 8.5 of orotic acid, orotidine, and orotidine 5'-phosphate with light at 254 nanometers yields the corresponding uracil derivative via the singlet excited state. This reaction completes a plausible prebiotic synthesis of uracil and its derivatives starting from HCN as the only carbon source.
Assuntos
Cianeto de Hidrogênio , Ácido Orótico , Descarboxilação , Ácido Orótico/análogos & derivados , Fotoquímica , Ribonucleotídeos , Uracila/síntese química , Uridina/análogos & derivados , Uridina/síntese química , Uridina Monofosfato/síntese químicaRESUMO
Malaria, caused by Plasmodia parasites, has re-emerged as a major problem, imposing its fatal effects on human health, especially due to multidrug resistance. In Plasmodia, orotidine 5'-monophosphate decarboxylase (ODCase) is an essential enzyme for the de novo synthesis of uridine 5'-monophosphate. Impairing ODCase in these pathogens is a promising strategy to develop novel classes of therapeutics. Encouraged by our recent discovery that 6-iodo uridine is a potent inhibitor of P. falciparum, we investigated the structure-activity relationships of various C6 derivatives of UMP. 6-Cyano, 6-azido, 6-amino, 6-methyl, 6- N-methylamino, and 6- N, N-dimethylamino derivatives of uridine were evaluated against P. falciparum. The mononucleotides of 6-cyano, 6-azido, 6-amino, and 6-methyl uridine derivatives were studied as inhibitors of plasmodial ODCase. 6-Azidouridine 5'-monophosphate is a potent covalent inhibitor of P. falciparum ODCase. 6-Methyluridine exhibited weak antimalarial activity against P. falciparum 3D7 isolate. 6- N-Methylamino and 6- N, N-dimethylamino uridine derivatives exhibited moderate antimalarial activities.
Assuntos
Antimaláricos/síntese química , Orotidina-5'-Fosfato Descarboxilase/antagonistas & inibidores , Plasmodium/efeitos dos fármacos , Uridina/análogos & derivados , Uridina/síntese química , Animais , Antimaláricos/farmacologia , Células CHO , Cricetinae , Cricetulus , Cristalografia por Raios X , Modelos Moleculares , Plasmodium/enzimologia , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/enzimologia , Plasmodium vivax/efeitos dos fármacos , Relação Estrutura-Atividade , Uridina/farmacologia , Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/síntese química , Uridina Monofosfato/farmacologiaRESUMO
Orotidine 5'-monophosphate decarboxylase (ODCase) has evolved to catalyze the decarboxylation of orotidine 5'-monophosphate without any covalent intermediates. Active site residues in ODCase are involved in an extensive hydrogen-bonding network. We discovered that 6-iodouridine 5'-monophosphate (6-iodo-UMP) irreversibly inhibits the catalytic activities of ODCases from Methanobacterium thermoautotrophicum and Plasmodium falciparum. Mass spectral analysis of the enzyme-inhibitor complex confirms covalent attachment of the inhibitor to ODCase accompanied by the loss of two protons and the iodo moiety. The X-ray crystal structure (1.6 A resolution) of the complex of the inhibitor and ODCase clearly shows the covalent bond formation with the active site Lys-72 [corrected] residue. 6-Iodo-UMP inhibits ODCase in a time- and concentration-dependent fashion. 6-Iodouridine, the nucleoside form of 6-iodo-UMP, exhibited potent antiplasmodial activity, with IC50s of 4.4 +/- 1.3 microM and 6.2 +/- 0.7 microM against P. falciparum ItG and 3D7 isolates, respectively. 6-Iodouridine 5'-monophosphate is a novel covalent inhibitor of ODCase, and its nucleoside analogue paves the way to a new class of inhibitors against malaria.
Assuntos
Antimaláricos/síntese química , Orotidina-5'-Fosfato Descarboxilase/antagonistas & inibidores , Uridina Monofosfato/análogos & derivados , Uridina/análogos & derivados , Animais , Antimaláricos/química , Antimaláricos/farmacologia , Células CHO , Cricetinae , Cricetulus , Cristalografia por Raios X , Espectrometria de Massas , Methanobacterium/enzimologia , Modelos Moleculares , Orotidina-5'-Fosfato Descarboxilase/química , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/enzimologia , Plasmodium falciparum/isolamento & purificação , Estereoisomerismo , Relação Estrutura-Atividade , Uridina/síntese química , Uridina/química , Uridina/farmacologia , Uridina Monofosfato/síntese química , Uridina Monofosfato/química , Uridina Monofosfato/farmacologiaRESUMO
The design and synthesis of novel 15-membered 11-azalides and 16-membered 11,12-diazalide starting from 16-membered macrolides are reported. A mobile linear dialdehyde was isolated via a cyclic tetraol which was prepared by osmium oxidation of a conjugated diene. One-pot macrocyclization of this dialdehyde with an amine or a diamine afforded corresponding 15-membered azalides or 11,12-diazalide. Fundamental SAR studies of 15-membered 11-azalides disclosed their potentiality as a lead molecule for further chemical modifications. For environmental preservation, sustainable chemistry for synthesis of these azalides is also discussed.
Assuntos
Antibacterianos/síntese química , Antibacterianos/farmacologia , Compostos Aza/síntese química , Azitromicina/análogos & derivados , Macrolídeos/síntese química , Macrolídeos/farmacologia , Antibacterianos/química , Compostos Aza/farmacologia , Azitromicina/química , Azitromicina/farmacologia , Bactérias/efeitos dos fármacos , Humanos , Kitasamicina/síntese química , Kitasamicina/farmacologia , Compostos Macrocíclicos , Macrolídeos/química , Testes de Sensibilidade Microbiana , Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/síntese química , Uridina Monofosfato/farmacologiaRESUMO
The increase in the number of bacterial strains resistant to known antibiotics is alarming. In this study we report the synthesis of novel compounds termed Lipophosphonoxins II (LPPO II). We show that LPPO II display excellent activities against Gram-positive and -negative bacteria, including pathogens and multiresistant strains. We describe their mechanism of action-plasmatic membrane pore-forming activity selective for bacteria. Importantly, LPPO II neither damage nor cross the eukaryotic plasmatic membrane at their bactericidal concentrations. Further, we demonstrate LPPO II have low propensity for resistance development, likely due to their rapid membrane-targeting mode of action. Finally, we reveal that LPPO II are not toxic to either eukaryotic cells or model animals when administered orally or topically. Collectively, these results suggest that LPPO II are highly promising compounds for development into pharmaceuticals.
Assuntos
Antibacterianos/química , Uridina Monofosfato/análogos & derivados , Animais , Antibacterianos/síntese química , Antibacterianos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Desenho de Fármacos , Farmacorresistência Bacteriana Múltipla , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Humanos , Bicamadas Lipídicas/química , Masculino , Camundongos Endogâmicos ICR , Testes de Sensibilidade Microbiana , Fosfolipídeos/química , Pirazóis/síntese química , Pirazóis/química , Pirazóis/farmacologia , Coelhos , Testes de Irritação da Pele , Estereoisomerismo , Relação Estrutura-Atividade , Uridina Monofosfato/síntese química , Uridina Monofosfato/química , Uridina Monofosfato/farmacologiaRESUMO
Inhibitors of orotidine monophosphate decarboxylase (ODCase) have applications in RNA viral, parasitic, and other infectious diseases. ODCase catalyzes the decarboxylation of orotidine monophosphate (OMP), producing uridine monophosphate (UMP). Novel inhibitors 6-amino-UMP and 6-cyano-UMP were designed on the basis of the substructure volumes in the substrate OMP and in an inhibitor of ODCase, barbituric acid monophosphate, BMP. A new enzyme assay method using isothermal titration calorimetry (ITC) was developed to investigate the inhibition kinetics of ODCase. The reaction rates were measured by monitoring the heat generated during the decarboxylation reaction of orotidine monophosphate. Kinetic parameters (k(cat) = 21 s(-1) and KM = 5 microM) and the molar enthalpy (DeltaH(app) = 5 kcal/mol) were determined for the decarboxylation of the substrate by ODCase. Competitive inhibition of the enzyme was observed and the inhibition constants (Ki) were determined to be 12.4 microM and 29 microM for 6-aza-UMP and 6-cyano-UMP, respectively. 6-Amino-UMP was found to be among the potent inhibitors of ODCase, having an inhibition constant of 840 nM. We reveal here the first inhibitors of ODCase designed by the principles of bioisosterism and a novel method of using isothermal calorimetry for enzyme inhibition studies.
Assuntos
Orotidina-5'-Fosfato Descarboxilase/antagonistas & inibidores , Orotidina-5'-Fosfato Descarboxilase/química , Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/síntese química , Calorimetria , Simulação por Computador , Desenho de Fármacos , Cinética , Modelos Moleculares , Termodinâmica , Uridina Monofosfato/químicaRESUMO
A series of UTP, UDP, and UMP derivatives and analogues were synthesized and evaluated at the human pyrimidinergic P2Y receptor subtypes P2Y2, P2Y4, and P2Y6 stably expressed in 1321N1 astrocytoma cells. Substituents at N3 of UTP were poorly tolerated by P2Y2 and P2Y4 receptors. In contrast, a large phenacyl substituent at N3 of UDP was well tolerated by the P2Y6 receptor, yielding a potent and selective P2Y6 receptor agonist (3-phenacyl-UDP, EC50=70 nM, >500-fold selective). The most potent and selective P2Y2 receptor agonist of the present series was 2-thio-UTP (EC50=50 nM, >or=30-fold selective vs P2Y4 and P2Y6). All modifications at the uracil base of UTP led to a decrease in potency at the P2Y4 receptor. A beta,gamma-dichloromethylene modification in the triphosphate chain of 5-bromo-UTP was tolerated by all three receptor subtypes, thus opening up a new strategy to obtain ectonucleotide diphosphohydrolase- and phosphatase-resistant P2Y2, P2Y4, and P2Y6 receptor agonists.
Assuntos
Agonistas do Receptor Purinérgico P2 , Nucleotídeos de Uracila/síntese química , Compostos Bicíclicos com Pontes/síntese química , Compostos Bicíclicos com Pontes/farmacologia , Linhagem Celular Tumoral , Humanos , Fosfatos de Inositol/biossíntese , Purinas/síntese química , Purinas/farmacologia , Receptores Purinérgicos P2 , Receptores Purinérgicos P2Y2 , Relação Estrutura-Atividade , Nucleotídeos de Uracila/farmacologia , Difosfato de Uridina/análogos & derivados , Difosfato de Uridina/síntese química , Difosfato de Uridina/farmacologia , Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/síntese química , Uridina Monofosfato/farmacologia , Uridina Trifosfato/análogos & derivados , Uridina Trifosfato/síntese química , Uridina Trifosfato/farmacologiaRESUMO
The pyrimidine acyclonucleoside benzyloxybenzyloxybenzylacyclouridine (BBBAU) showed growth inhibitory activity against the human colon cancer HCT-8 cell line with a 50% inhibitory concentration of 55 microM. Unlike its parent compounds, BBBAU was an extremely weak inhibitor of uridine phosphorylase. This acyclonucleoside analogue is an inhibitor of thymidylate synthase (TS) as determined by inhibition of [6-3H]-2'-deoxyuridine incorporation into DNA, inhibition of 3H release from [5-3H]-2'-deoxyuridine, and decrease in both the free and total TS 5'-fluoro-2'-deoxyuridine 5'-monophosphate binding sites. Kinetic analysis revealed that BBBAUMP, the monophosphate analogue of BBBAU, is a competitive inhibitor of purified human recombinant TS with a Ki of 8.0 microM. Nucleoside transport and uptake studies revealed that BBBAU (30 microM) inhibited the initial rate of transport and the total uptake of thymidine (25 microM). In contrast, while BBBAU (30 microM) inhibited the initial rate of transport of 5-fluoro-2'-deoxyuridine (FdUrd, 25 microM), its intracellular accumulation was increased. BBBAU (10 and 50 microM, respectively) potentiated FdUrd growth inhibition of HCT-8 cells and significantly enhanced the cytotoxic effects of FdUrd (0.3 and 1 microM, respectively) against HCT-8 cells using a clonogenic assay system. This combination resulted in additive inhibitory effects on TS activity resulting in greater depletion of dTTP pools. Moreover, the incorporation of radiolabeled FdUrd into the DNA fraction of HCT-8 cells was enhanced. The potential importance of this novel combination for human colon cancer chemotherapy is discussed.
Assuntos
Antimetabólitos Antineoplásicos/farmacologia , Divisão Celular/efeitos dos fármacos , Floxuridina/farmacologia , Uracila/análogos & derivados , Linhagem Celular , Neoplasias do Colo , Replicação do DNA/efeitos dos fármacos , Desoxiuridina/metabolismo , Sinergismo Farmacológico , Floxuridina/metabolismo , Humanos , Cinética , Proteínas Recombinantes/antagonistas & inibidores , Timidilato Sintase/antagonistas & inibidores , Uracila/farmacologia , Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/síntese química , Uridina Monofosfato/farmacologiaRESUMO
The synthesis of an 8-deazafolate analogue of the intermediate in the methylation of 2'-deoxyuridylate is described. Alkylation of diethyl 5,6,7,8-tetrahydro-8-deazafolate with 3'-O-acetyl-5-(bromomethyl)-2'-deoxyuridine 5'-[bis-(trichlorethyl) phosphate], followed by removal of the trichloroethyl groups with a Zn/Cu couple and mild saponification, gave the target inhibitor N-[4-[[[2-amino-3,4,5,6,7, 8-hexahydro-4-oxo-5-[(2'-deoxyuridin-5-yl)methyl]-pyrido[3,2-d] pyrimidin-6-yl]methyl]amino]benzoyl]-L-glutamic acid 5'-monophosphate. The free nucleoside and the 5'-(methyl phosphate) diester were similarly prepared. Each of these reactions yielded a pair of diastereoisomers about C-6 of the reduced deazafolate in approximately a 1:1 ratio. These diastereoisomeric mixtures were evaluated as inhibitors of thymidylate synthetase derived from human tumor (HeLa) cells. The 5'-monophosphate was a potent inhibitor, competitive with respect to both 2'-deoxyuridylate (Ki = 0.06 microM) and tetrahydrofolate (Ki = 0.25 microM). In contrast, the nucleoside and the nucleotide methyl ester were poorer inhibitors by more than 3 orders of magnitude, attesting to the importance of the anionic function at the nucleoside 5'-position in the affinity of an inhibitor for the enzyme active site.
Assuntos
Metiltransferases/antagonistas & inibidores , Timidilato Sintase/antagonistas & inibidores , Nucleotídeos de Uracila/síntese química , Uridina Monofosfato/síntese química , Células HeLa/enzimologia , Humanos , Indicadores e Reagentes , Cinética , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Ligação Proteica , Espectrofotometria Ultravioleta , Relação Estrutura-Atividade , Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/farmacologiaRESUMO
Condensation of 6-azauridine with ethyl levulinate, followed by saponification or phosphorylation, leads to 2',3'-O-[1-(2-carboxyethyl)ethylidene]-6-azauridine and its 5'-monophosphate. The latter was coupled to 6-aminohexylagarose via its carboxylic group. Using the same synthetic route, agarose-linked uridine 5'-monophosphate has been prepared. Both polymers show specific binding toward orotidine-5'-monophosphate decarboxylase. The immobilized inhibitor (6-azauridine 5'-monophosphat) binds the enzyme more strongly than the immobilized uridine 5'-monophosphate. Both resins have been used to separate orotidine-5'-monophosphate decarboxylase from orotidine-5'-monophosphate pyrophosphorylase.
Assuntos
Azauridina/análogos & derivados , Carboxiliases/antagonistas & inibidores , Orotidina-5'-Fosfato Descarboxilase/antagonistas & inibidores , Resinas Sintéticas/síntese química , Nucleotídeos de Uracila/síntese química , Uridina Monofosfato/síntese química , Azauridina/síntese química , Azauridina/farmacologia , Cromatografia de Afinidade , Orotidina-5'-Fosfato Descarboxilase/metabolismo , Resinas Sintéticas/farmacologia , Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/farmacologiaRESUMO
A series of 5-alkyl-2'-deoxyuridine 3',5'-cyclic monophosphates (5-R-cdUMP's, R = Et, i-Pr, n-Pr, n-Bu, n-Pent, n-Hex, n-Oct) was prepared and tested in culture systems as antitumor and antiviral agents in comparison to the 5-alkyl-2'-deoxyuridines (5-R-dUrd's) themselves. Only the 5-Et- and 5-n-Bu-cdUMP showed appreciable cytostatic activities against murine L1210 and human lymphoblast Raji cells (ID50 range: 28-82 micrograms/mL). 5-Et-dUrd itself was much more active (ID50 = 1.6 and 2.9 micrograms/mL). The 5-i-Pr-, and 5-n-Bu-dUrd's were inactive, but activity increased again for groups with chain lengths of five carbons or greater. 5-Et-cdUMP and 5-Et-dUrd had greatly reduced activities against deoxythymidine kinase deficient (TK-) L1210 and Raji cells. 5-Et-cdUMP evidently is not an efficient prodrug source of the corresponding 5'-monophosphate where the TK- cells are concerned. Of the 5-R-cdUMP's, 5-Et-cdUMP displayed reasonably good antiviral potency against herpes simplex types 1 and 2 (MIC50, mostly 7-70 micrograms/mL) and vaccinia virus (MIC, 70 micrograms/mL). The activity was nonetheless 10- to 100-fold less than that for 5-Et-dUrd. The other 5-R-dUrd's generally showed decreasing antiviral activity with increasing 5-R chain length. Methyl and/or benzyl neutral triesters of certain 5-R-cdUMP's were inactive as antivirals and largely inactive against tumor cells in culture. In contrast to the 5'-monophosphates, the 5-R-cdUMP's failed to inhibit thymidylate synthetase from L1210 cells.
Assuntos
Antineoplásicos/síntese química , Antivirais/síntese química , Nucleotídeos Cíclicos/síntese química , Nucleotídeos de Uracila/síntese química , Uridina Monofosfato/síntese química , Animais , Antineoplásicos/farmacologia , Antivirais/farmacologia , Linhagem Celular , Humanos , Leucemia L1210/tratamento farmacológico , Espectroscopia de Ressonância Magnética , Camundongos , Nucleotídeos Cíclicos/farmacologia , Relação Estrutura-Atividade , Timidina Quinase/análise , Timidilato Sintase/antagonistas & inibidores , Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/farmacologiaRESUMO
A novel approach to the intracellular delivery of nucleotides using phosphoramidate-based prodrugs is described. Specifically, we have developed phosphoramidate prodrugs of the anticancer nucleotide 5-fluoro-2'-deoxyuridine-5'monophosphate (FdUMP). These phosphoramidate prodrugs contain an ester group that undergoes intracellular activation liberating phosphoramidate anion, which undergoes spontaneous cyclization and P-N bond cleavage to yield the nucleoside monophosphate quantitatively. In vitro evaluation of 5-fluoro-2'-deoxyuridine phosphoramidate prodrugs 2a and 3b against L1210 mouse leukemia cells show potent inhibition of cell growth (IC(50) 0.5-3 nM). Cell-based thymidylate synthase inhibition studies show that, in contrast to FUdR, the nitrofuran compound 2a is of comparable potency in wild type vs thymidine kinase deficient LM cells. This result indicates that the activation of this novel prodrug occurs via the proposed mechanism of intracellular delivery. However, naphthoquinone 3b has an IC(50) value for thymidylate synthase inhibition that is comparable to FUdR in thymidine kinase deficient cells. Further studies revealed that 3b rapidly decomposes to the nucleotide in cell culture medium, suggesting that the naphthoquinone analogue is not sufficiently stable to function as a nucleotide prodrug.
Assuntos
Antineoplásicos/síntese química , Fluordesoxiuridilato/análogos & derivados , Fluordesoxiuridilato/síntese química , Pró-Fármacos/síntese química , Uridina Monofosfato/síntese química , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Ensaios de Seleção de Medicamentos Antitumorais , Inibidores Enzimáticos/farmacologia , Floxuridina/farmacologia , Fluordesoxiuridilato/química , Fluordesoxiuridilato/farmacologia , Espectroscopia de Ressonância Magnética , Camundongos , Pró-Fármacos/química , Pró-Fármacos/farmacologia , Timidina Quinase/deficiência , Timidilato Sintase/antagonistas & inibidores , Células Tumorais Cultivadas , Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/química , Uridina Monofosfato/farmacologiaRESUMO
A synthesis of 6-formyluridine 5'-monophosphate (6-formylUMP) in 6 steps starting from uridine is described. This approach should be applicable to the preparation of other O5'-phosphorylated 6-formylUrds such as 6-formylUDP and 6-formylUTP.
Assuntos
Uridina Monofosfato/análogos & derivados , Uridina Monofosfato/síntese químicaRESUMO
Modern studies now favor the fact that extraterrestrial organic molecules served as an important source of biological important substances on the primitive Earth. It is presumed that these space-made organic molecules could be transported safely to the Earth surface being associated with mineral grains. It is important to test whether nucleotides synthesized in Earth orbit could be protected by lunar surface regolite. The phosphorylation of adenosine, uridine and thymidine has been studied with respect of their further transformations and degradation in presence of mineral bed. After retrieval, HPLC analysis is used to identify all the mononucleotides of certain nucleosides. It has been shown, that exposure of the investigated nucleosides as dry films in space conditions in the presence of Lunar soil increases the yield of synthesized nucleotides in 1.1-3.0 times as compared with the exposure of the same samples in absence of Lunar soil. To identify and evaluate the principal source of energy in open space responsible for nucleotide synthesis reaction laboratory experiments were performed. It has been shown, that vacuum ultra violet (VUV 145 nm) radiation promotes nucleotide synthesis more effectively than ultra violet (UV 254 nm) while the presence of Lunar soil increases reaction yield in 1.5-2.0 times. Formation of 5'-mononucleotides seemed to be the most effective reaction both in flight and in laboratory experiments. Protective action of lunar soil on synthesized nucleotides against UV radiation has been shown in open Space conditions.
Assuntos
Evolução Química , Nucleotídeos/síntese química , Solo/análise , Voo Espacial , Raios Ultravioleta , Monofosfato de Adenosina/síntese química , Planeta Terra , Exobiologia , Meio Ambiente Extraterreno , Lua , Timidina Monofosfato/síntese química , Uridina Monofosfato/síntese químicaRESUMO
Diiminosuccinonitrile (DISN) has been investigated as a potential prebiotic phosphorylating agent. It is formed readily by the oxidation of diaminomaleonitrile (DAMN), a tetramer of HCN. DISN effects the cyclization of 3'-adenosine monophosphate to adenosine 2',3'-cyclic phosphace in up to 40% yield. The DISN-mediated phosphorylation of uridine to uridine mono-phosphate does not proceed efficiently in aqueous solution. The reaction of DISN and BrCN with uridine-5'-phosphate and uridine results in the formation of 2,2'-anhydronucleotides and 2,2'-anhydronucleosides respectively, and other reaction products resulting from an initial reaction at the 2'- and 3'-hydroxyl groups. The clay mineral catalysis of the cyclization of adenosine-3'-phosphate was investigated using homoionic montmorillonites.
Assuntos
Monofosfato de Adenosina/química , Evolução Química , Nitrilas/química , Nucleosídeos/química , Nucleotídeos/síntese química , Uridina/química , Bentonita , Catálise , Evolução Molecular , Nucleosídeos/síntese química , Nucleotídeos/química , Fosforilação , Uridina Monofosfato/síntese química , Uridina Monofosfato/químicaRESUMO
A chromophorics and fluorescent analog of uridine 5'-monophosphate (UMP), a known competitive inhibitor of UDPglucose 4-epimerase was synthesised. This analog, namely 2',3'-O-(2,4,6-trinitrocyclohexadienylidene) uridine 5'-monophosphate, was found to be a powerful reversible inhibitor of UDPglucose 4-epimerase indicating its interaction with the substrate binding site of the enzyme. The extreme sensitivity of the fluorescence emission spectrum of this analog to solvent polarity makes it an excellent probe for the study of the environment at the active site of the enzyme. We report here the effective use of this UMP analog to demonstrate that the hydroxyl groups of the ribose moiety of UMP and presumably the substrates (UDPgalactose and UDPglucose) do not reside in a hydrophobic milieu.