RESUMO
Fragile histidine triad (HIT) proteins (Fhits) occur in all eukaryotes but their function is largely unknown. Human Fhit is presumed to function as a tumour suppressor. Previously, we demonstrated that Fhits catalyse hydrolysis of not only dinucleoside triphosphates but also natural adenosine 5'-phosphoramidate (NH2-pA) and adenosine 5'-phosphosulfate (SO4-pA) as well as synthetic adenosine 5'-phosphorofluoridate (F-pA). In the present study, we describe an Fhit-catalysed displacement of the amino group of nucleoside 5'-phosphoramidates (NH2-pNs) or the sulfate moiety of nucleoside 5'-phosphosulfates (SO4-pNs) by fluoride anion. This results in transient accumulation of the corresponding nucleoside 5'-phosphorofluoridates (F-pNs). Substrate specificity and kinetic characterization of the fluorolytic reactions catalysed by the human Fhit and other examples of involvement of fluoride in the biochemistry of nucleotides are described. Among other HIT proteins, human histidine triad nucleotide-binding protein (Hint1) catalysed fluorolysis of NH2-pA 20 times and human Hint2 40 times more slowly than human Fhit.
Assuntos
Hidrolases Anidrido Ácido/metabolismo , Monofosfato de Adenosina/análogos & derivados , Adenosina Fosfossulfato/metabolismo , Fluoretos/metabolismo , Proteínas de Neoplasias/metabolismo , Fosfatos/metabolismo , Monofosfato de Adenosina/metabolismo , Catálise , Humanos , Cinética , Estrutura Molecular , Especificidade por SubstratoRESUMO
This chapter provides an overview of recent advances in the development of new methods and protocols for the formation of the P-C bond using H-phosphonate diesters as starting materials. Various chemical and stereochemical aspects of the transition metal-catalyzed cross-coupling and organocatalyst-promoted reactions which are relevant to the synthesis of structurally diverse C-phosphonate derivatives are surveyed.
RESUMO
This review covers recent progress in the preparation of H-phosphonate mono- and diesters, basic studies on mechanistic and stereochemical aspects of this class of phosphorus compounds, and their fundamental chemistry in terms of transformation of P-H bonds into P-heteroatom bonds. Selected recent applications of H-phosphonate derivatives in basic organic phosphorus chemistry and in the synthesis of biologically important phosphorus compounds are also discussed.
Assuntos
Organofosfonatos/síntese química , Fósforo/química , Ésteres , Nucleosídeos/química , Oligonucleotídeos/química , Compostos Organofosforados/química , EstereoisomerismoRESUMO
Nucleotides, their analogues, and other phosphate esters and phosphoramidates often contain the triethylammonium cation as a counterion. We found that this may be lost during chromatographic purification or concentration of solutions, yielding products in acidic forms or containing sub-stoichiometric amounts of the counterion. This in turn may be detrimental, e.g., due to possible decomposition of a compound or inaccurate sample preparation. Correlations between the structure of studied compounds and their susceptibility for cation loss were analyzed. Modifications in preparative techniques were developed to obtain the studied compounds with stoichiometric anion to cation ratios.
Assuntos
Nucleotídeos/análise , Compostos de Amônio Quaternário/químicaRESUMO
In this review a short account of our work on the synthesis and biological activity of electrically neutral and charged anti-HIV and anticancer pronucleotides, presented on the background of the contemporary research in this area, is given.
RESUMO
Di-aryl nucleoside phosphotriesters have been explored as a new type of pronucleotides for the purpose of anti-HIV-1 therapy and efficient synthetic protocols, based on H-phosphonate chemistry, have been developed for the preparation of this class of compounds. It was found that anti-HIV-1 activity of the phosphotriesters bearing an antiviral nucleoside moiety (AZT, ddA) and also ddU was due, at least partially, to intracellular conversion into the corresponding nucleoside 5'-monophosphates, and their efficiency correlated well with the pK(a) values of the aryloxy groups present.
Assuntos
Fármacos Anti-HIV/síntese química , Nucleosídeos/síntese química , Nucleotídeos/síntese química , Organofosfonatos/síntese química , Fármacos Anti-HIV/química , Fármacos Anti-HIV/farmacologia , Linhagem Celular , Células Cultivadas , HIV/fisiologia , Humanos , Hidroxiácidos/síntese química , Hidroxiácidos/química , Hidroxiácidos/farmacologia , Testes de Sensibilidade Microbiana , Estrutura Molecular , Nucleosídeos/química , Nucleosídeos/farmacologia , Nucleotídeos/química , Nucleotídeos/farmacologia , Organofosfonatos/química , Organofosfonatos/farmacologia , Replicação Viral/efeitos dos fármacosRESUMO
We have designed a new type of AZT and ddU phosphoramidate diesters containing various combinations of 2-, 3-, 4-aminopyridine and 2-, 3-, 4-hydroxypyridine moieties attached to the phosphorus center, as potential anti-HIV pronucleotides. Depending on the pKa values of the aminopyridines and the hydroxypyridines used, alternative synthetic strategies based on H-phosphonate chemistry were developed for their preparation. Synthetic aspects of these transformations and the biological activity of the synthesized compounds are discussed.
Assuntos
Amidas/farmacologia , Fármacos Anti-HIV/química , Desenho de Fármacos , Organofosfonatos/uso terapêutico , Ácidos Fosfóricos/farmacologia , Amidas/síntese química , Amidas/química , Aminopiridinas , Fármacos Anti-HIV/síntese química , Didesoxinucleosídeos , Organofosfonatos/química , Ácidos Fosfóricos/síntese química , Ácidos Fosfóricos/química , Piridinas , ZidovudinaRESUMO
We have designed and synthesized new 5-fluoro-2'-deoxyuridine 5'-phosphate pronucleotides which can function as potential agents against the glioblastoma multiforme tumor. Their anti-malignant potency has been tested against T98G, U-118â¯MG, U-87â¯MG gliomas, HeLa, and Caco-2 cancer cell lines, using MRC-5 healthy cells as a reference. Five of the sixteen compounds (4c, 4f-i) exhibited significant anticancer potency and high selectivity indices (SI 12-66). It is likely that these zwitterionic pronucleotides may function in a similar manner to zwitterionic phospholipids, by inducing cell membrane charge disorder, making the cell permeable to bioactive agents. The most promising therapeutic pronucleotides 4c, 4f-h, have high intestinal-blood uptake potency (Caco-2â¯cell line), and may be considered as potential, orally administrated, anticancer drugs.
Assuntos
Antineoplásicos/farmacologia , Monofosfato de Citidina/análogos & derivados , Glioblastoma/tratamento farmacológico , Nucleotídeos/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Monofosfato de Citidina/química , Monofosfato de Citidina/farmacologia , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Glioblastoma/patologia , Humanos , Estrutura Molecular , Nucleotídeos/síntese química , Nucleotídeos/química , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
A configuration of ligands around a phosphorus atom in P-chiral dinucleoside monophosphate analogues can be described using DP/LP stereochemical notation, which allows immediate correlation between the notation of configuration and the actual spatial arrangement of the phosphorus ligands. The area of applications of this new stereochemical nomenclature covers dinucleoside units bridged by virtually any type of tri-and tetra-coordinated phosphorus moieties, that is, phosphorothioates, phosphoramidates, phosphoramidites, boranephosphates, methanephosphonates, H-phosphonates, and many others.
Assuntos
Química/métodos , Fosfatos de Dinucleosídeos/química , Nucleosídeos/química , Nucleotídeos/química , Terminologia como Assunto , Ligantes , Modelos Químicos , Modelos Moleculares , Estrutura Molecular , Oxigênio/química , Proteínas Recombinantes de Fusão/química , EstereoisomerismoRESUMO
Recently, we have proposed a new DP/LP stereochemical notation for P-chiral dinucleoside monophosphate analogues that permits simple correlation between spatial arrangement of the substituents and the configuration at the phosphorus center. As an extension of this work, we present here applications of the DP/LP notation to derivatives containing only one nucleoside unit (e.g., alkyl nucleoside phosphodiesters, nucleoside phosphomonoesters, cyclic phosphate derivatives, nucleoside di-, and triphosphates) and to nonnucleosidic phosphorus compounds.
Assuntos
Bioquímica/métodos , Nucleosídeos/química , Nucleotídeos/química , Fósforo/química , Terminologia como Assunto , Ligantes , Modelos Químicos , Estrutura Molecular , EstereoisomerismoRESUMO
Several ribonucleoside analogues with modifications in the nucleobase and sugar moiety have been screened for anti-glioma activity in the T98G glioma cell line using cervical (HeLa) cell line as reference human malignant cells, and lung fibroblast (MCR-5) cell line as non-cancerous reference cells. Among the investigated compounds, ribonucleosides containing 6-chloropurine (3), 7-guanine (5) and a pyrrolopyrimidine (18) as nucleobases, show promising anti-glioma activity with good selectivity indices, and can be considered as lead structures for further anti-cancer studies.
Assuntos
Adenosina/análogos & derivados , Adenosina/farmacologia , Antineoplásicos/farmacologia , Citidina/análogos & derivados , Citidina/farmacologia , Guanosina/análogos & derivados , Guanosina/farmacologia , Neoplasias Encefálicas/tratamento farmacológico , Ensaios de Seleção de Medicamentos Antitumorais , Furanos/farmacologia , Glioblastoma/tratamento farmacológico , Células HeLa , Humanos , Concentração Inibidora 50RESUMO
New aromatic and aliphatic 3'-O-acyl-5-fluoro-2'-deoxyuridine derivatives were synthesized and evaluated as candidates for prodrugs against various cancer cell lines. As the most promising candidate for antimalignant therapeutics was found a dual-acting acyl derivative 7h, which apparently released not only the known anticancer nucleoside, 5-fluoro-2'-deoxyuridine (FdU), but also an additional active metabolite, acetylsalicylic acid, reinforcing thus therapeutic effect of FdU. Promising therapeutic indices showed also some aromatic dicarboxylic acids derivatives decorated with FdU esters (11 and 12).
Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Desoxiuridina/análogos & derivados , Pró-Fármacos/farmacologia , Antineoplásicos/síntese química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Desoxiuridina/síntese química , Desoxiuridina/farmacologia , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Pró-Fármacos/síntese química , Pró-Fármacos/química , Relação Estrutura-AtividadeRESUMO
A new stereochemical notation for P-chiral nucleotide analogues and related compounds is proposed In this notation, the names of configurations, designated as D(P) and L(P), are derived from a geometrical relationship, rather than from priority rules, of substituents at the phosphorus centre. This new stereochemical description offers clear advantages over the CIP R/S nomenclature, particularly when used for comparing the influence of absolute configuration at the phosphorus centre on physicochemical and biological properties of oligonucleotide analogues or in stereochemical correlation analysis of P-chiral nucleotide derivatives.
Assuntos
Nucleotídeos/química , Terminologia como Assunto , Bioquímica/métodos , Estrutura Molecular , EstereoisomerismoRESUMO
Sixteen diribonucleoside (3'-5')-H-phosphonates were synthesized via condensation of the protected ribonucleoside 3'-H-phosphonates with nucleosides, and the influence of a nucleoside sequence on the observed stereoselectivity was analyzed. 31P NMR spectroscopy was used to evaluate a relationship between chemical shift and absolute configuration at the phosphorous center of the H-phosphonate diesters as well as of the corresponding phosphorothioate diesters. Although for the most cases such correlation was found, there was however several exceptions to the rule where the relative positions of resonances arisingfrom Rp and Sp diastereomers were reversed.
Assuntos
Fosfatos de Dinucleosídeos/síntese química , Compostos Organofosforados/química , Fosfatos de Dinucleosídeos/química , Espectroscopia de Ressonância Magnética , Isótopos de Fósforo/química , EstereoisomerismoRESUMO
It was found that in stereoselective condensations of ribonucleoside 3'-H-phosphonates with alcohols, the major diastereomer of the produced H-phosphonate diesters is formed from the minor diastereomer of the intermediate phosphonic-pivalic anhydride.
Assuntos
Biologia Molecular/métodos , Nucleosídeos/química , Organofosfonatos/química , Espectroscopia de Ressonância Magnética , Modelos Químicos , Conformação Molecular , Nucleotídeos/química , EstereoisomerismoRESUMO
Stereoselectivity in condensation of protected ribonucleoside 3'-H-phosphonates with hydroxylic components was investigated using 31P NMR spectroscopy. The correlation between absolute configuration at the phosphorus center and the chemical shifts of the produced H-phosphonate diesters and the corresponding phosphorothioates, was studied.
Assuntos
Espectroscopia de Ressonância Magnética/métodos , Espectrofotometria/métodos , Etanol/química , Guanosina/química , Modelos Químicos , Conformação Molecular , Conformação de Ácido Nucleico , Oligonucleotídeos/química , Organofosfonatos/química , Fósforo , Isótopos de Fósforo/química , Prótons , Solventes/química , EstereoisomerismoRESUMO
In this paper a short account of our recent basic studies aiming toward development of new synthetic methods for the preparation of nucleotide analogues using H-phosphonate chemistry is presented.
Assuntos
Nucleotídeos/síntese química , Organofosfonatos/química , Compostos de Fósforo/síntese química , Fósforo/química , Modelos Químicos , Nucleosídeos/química , Nucleotídeos/química , EstereoisomerismoRESUMO
Recently, AZT (N-pyridyl)phosphoramidates were reported as a new type of potential anti-HIV therapeutics. In continuation of that work, here we present new (N-heteroaryl)phosphoramidate derivatives of antiviral 2',3'-dideoxynucleosides containing other types of N-heteroaryl moieties, particularly those with higher lipophilicity. The present studies comprise mechanistic investigations using (31)P NMR correlation analysis, which permitted improvements in the synthetic procedures. The obtained compounds were tested in biological systems to establish their cytotoxicity and anti-HIV activity. The results were analyzed with respect to possible correlations between biological and physico-chemical properties of the phosphoramidates studied, to get some insight into their antiviral mode of action.
Assuntos
Fármacos Anti-HIV/farmacologia , Didesoxinucleosídeos/farmacologia , Fármacos Anti-HIV/síntese química , Fármacos Anti-HIV/química , Linhagem Celular , Sobrevivência Celular , Didesoxinucleosídeos/síntese química , Didesoxinucleosídeos/química , Relação Dose-Resposta a Droga , HIV-1/efeitos dos fármacos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Testes de Sensibilidade Microbiana , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
[reaction: see text] Aryl nucleoside H-phosphonates 3 and aryl nucleoside P-acylphosphonates 4, generated in situ from the appropriate H-phosphonate 1 and acylphosphonate monoesters 2, respectively, reacted rapidly in the presence of tertiary amines to produce in high yields the extended, pyrophosphate analogues, diaryl dinucleoside phosphonate-phosphate derivatives 6. These, depending on a substituent on the alpha-carbon of the phosphonate moiety, either underwent transformation into the dinucleoside phosphonate-phosphate 7 or afforded nucleoside H-phosphonates 8 and aryl nucleoside phosphate 9.
Assuntos
Nucleosídeos/química , Nucleotídeos/síntese química , Organofosfatos/química , Organofosfonatos/química , Nucleotídeos/químicaRESUMO
Chemoselectivity and stereospecificity of iodine mediated oxidative couplings using separate diastereomers of dinucleoside H-phosphonate and H-phosphonothioate with various N- and O-binucleophiles were investigated.