RESUMEN
The L-enantiomer of 4'-C-ethynyl-2'-deoxycytidine (2) was synthesized, but did not show any activity against HIV-1 up to 100 microM.
Asunto(s)
Desoxicitidina/síntesis química , Concentración de Iones de Hidrógeno , Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacología , Línea Celular , Colorantes , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacología , VIH-1/efectos de los fármacos , Indicadores y Reactivos , Espectroscopía de Resonancia Magnética , Espectrometría de Masa Bombardeada por Átomos Veloces , Estereoisomerismo , Sales de Tetrazolio , TiazolesRESUMEN
A series of 4'-ethynyl (4'-E) nucleoside analogs were designed, synthesized, and identified as being active against a wide spectrum of human immunodeficiency viruses (HIV), including a variety of laboratory strains of HIV-1, HIV-2, and primary clinical HIV-1 isolates. Among such analogs examined, 4'-E-2'-deoxycytidine (4'-E-dC), 4'-E-2'-deoxyadenosine (4'-E-dA), 4'-E-2'-deoxyribofuranosyl-2,6-diaminopurine, and 4'-E-2'-deoxyguanosine were the most potent and blocked HIV-1 replication with 50% effective concentrations ranging from 0.0003 to 0.01 microM in vitro with favorable cellular toxicity profiles (selectivity indices ranging 458 to 2,600). These 4'-E analogs also suppressed replication of various drug-resistant HIV-1 clones, including HIV-1(M41L/T215Y), HIV-1(K65R), HIV-1(L74V), HIV-1(M41L/T69S-S-G/T215Y), and HIV-1(A62V/V75I/F77L/F116Y/Q151M). Moreover, these analogs inhibited the replication of multidrug-resistant clinical HIV-1 strains carrying a variety of drug resistance-related amino acid substitutions isolated from HIV-1-infected individuals for whom 10 or 11 different anti-HIV-1 agents had failed. The 4'-E analogs also blocked the replication of a non-nucleoside reverse transcriptase inhibitor-resistant clone, HIV-1(Y181C), and showed an HIV-1 inhibition profile similar to that of zidovudine in time-of-drug-addition assays. The antiviral activity of 4'-E-thymidine and 4'-E-dC was blocked by the addition of thymidine and 2'-deoxycytidine, respectively, while that of 4'-E-dA was not affected by 2'-deoxyadenosine, similar to the antiviral activity reversion feature of 2',3'-dideoxynucleosides, strongly suggesting that 4'-E analogs belong to the family of nucleoside reverse transcriptase inhibitors. Further development of 4'-E analogs as potential therapeutics for infection with multidrug-resistant HIV-1 is warranted.
Asunto(s)
Fármacos Anti-VIH/farmacología , Desoxirribonucleósidos/farmacología , VIH-1/efectos de los fármacos , Fármacos Anti-VIH/antagonistas & inhibidores , Desoxirribonucleósidos/antagonistas & inhibidores , Interacciones Farmacológicas , Resistencia a Múltiples Medicamentos/fisiología , Estabilidad de Medicamentos , Infecciones por VIH/virología , VIH-1/aislamiento & purificación , VIH-2/efectos de los fármacos , Humanos , Pruebas de Sensibilidad MicrobianaRESUMEN
4'-C-Ethynyl-beta-D-arabino- and 4'-C-ethynyl-2'-deoxy-beta-D-ribo-pentofuranosylpyrimidine and -purine nucleosides were synthesized and evaluated for their in vitro anti-HIV activity. The key intermediate, 4-C-ethynyl- or 4-C-triethylsilylethynyl-D-ribo-pentofuranose, was prepared from D-glucose and glycosidated with various pyrimidine or purine bases. The arabinopyrimidine derivatives were prepared from the corresponding ribo derivatives via O(2),2'-anhydro nucleosides. The 2'-deoxy-ribo derivatives were synthesized by radical reduction of 2'-bromo or 2'- phenoxythiocarbonyloxy nucleosides. Among these 4'-C-ethynyl nucleosides, seven analogues proved to be potent against HIV-1 in vitro with EC(50) values ranging from 0.0003 to 0. 03 microM. These compounds also exerted activity against clinical and multi-dideoxy-nucleoside-resistant HIV-1 strains with comparable EC(50) values. Three such 4'-C-ethynyl-2'-deoxypurine analogues including 4'-C-ethynyl-2'-deoxyadenosine and 4'-C-ethynyl-2, 6-diamino-2'-deoxypurine were less cytotoxic [selectivity indices (SIs): 975-2733] than three 4'-C-ethynyl-2'-deoxycytidine analogues (SIs: 63-363). 4'-C-Ethynyl-5-fluoro-2'-deoxycytidine was least toxic (SI: >3333) and potent against all HIV strains tested.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Nucleósidos de Purina/síntesis química , Nucleósidos de Pirimidina/síntesis química , Fármacos Anti-VIH/química , Fármacos Anti-VIH/farmacología , Línea Celular , Farmacorresistencia Microbiana , VIH-1/efectos de los fármacos , VIH-1/aislamiento & purificación , Nucleósidos de Purina/química , Nucleósidos de Purina/farmacología , Nucleósidos de Pirimidina/química , Nucleósidos de Pirimidina/farmacología , Relación Estructura-ActividadRESUMEN
Searching for more effective anti-HIV agents, we have prepared 4'-ethynyl-purine nucleosides. They were derived in several steps from 4-C-triethylsilylethynyl ribose, which was used as an intermediate in the synthesis of pyrimidine nucleosides. The adenine derivative exhibited significant anti-HIV activity and favorable cytotoxicity profile in vitro.
Asunto(s)
Fármacos Anti-VIH/química , Fármacos Anti-VIH/síntesis química , Nucleósidos de Purina/química , Nucleósidos de Purina/síntesis química , Fármacos Anti-VIH/farmacología , Fármacos Anti-VIH/toxicidad , Muerte Celular/efectos de los fármacos , Línea Celular , Diseño de Fármacos , VIH/efectos de los fármacos , Humanos , Técnicas In Vitro , Métodos , Nucleósidos de Purina/farmacología , Nucleósidos de Purina/toxicidad , Relación Estructura-ActividadRESUMEN
4'-C-Ethynyl-beta-D-arabino-pentofuranosyl thymine (14) and cytosine (16), and 4'-C-ethynyl-2'-deoxy-beta-D-ribo-pentofuranosyl thymine (25) and cytosine (27) were synthesized by properly protected 4'-C-hydroxy-methyl-3,5-di-O-benzyl-alpha-D-ribo-pentofuranose (1) from D-glucose. Among them, 2'-deoxy derivatives 25 and 27 exhibited antiviral activity, while cytidine derivatives 16 and 27 inhibited the growth of neoplastic cells.
Asunto(s)
Antineoplásicos/síntesis química , Antivirales/síntesis química , Nucleósidos/síntesis química , Pirimidinas/síntesis química , Antineoplásicos/farmacología , Antivirales/farmacología , Humanos , Leucemia Experimental/tratamiento farmacológico , Inhibidores de la Síntesis del Ácido Nucleico/síntesis química , Inhibidores de la Síntesis del Ácido Nucleico/farmacología , Nucleósidos/farmacología , Pirimidinas/farmacología , Simplexvirus/efectos de los fármacos , Células Tumorales CultivadasRESUMEN
4'-C-Ethynyl-beta-D-arabino- and 4'-C-ethynyl-beta-D-2'-deoxy-ribo-pentofuranosyl pyrimidines were synthesized. Most of these 4'-ethynyl nucleosides showed interesting biological activities.
Asunto(s)
Antivirales/síntesis química , Nucleósidos/síntesis química , Animales , Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacología , Antimetabolitos Antineoplásicos/síntesis química , Antimetabolitos Antineoplásicos/toxicidad , Antivirales/farmacología , División Celular/efectos de los fármacos , Línea Celular , Diseño de Fármacos , VIH/efectos de los fármacos , Herpesvirus Humano 1/efectos de los fármacos , Herpesvirus Humano 2/efectos de los fármacos , Humanos , Indicadores y Reactivos , Células KB , Leucemia P388 , Ratones , Nucleósidos/farmacologíaRESUMEN
The 4'-C-ethynyl-ß-D-ribo-pentofuranosylpyrimidines were prepared from D-glucose through properly protected 4'-C-formyl-D-ribo-ribofuranose as the key intermediate, and preliminary biological tests against some viruses and tumor cells showed that the compounds were not active.
RESUMEN
The effects of four kinds of 2'-deoxy-L-nucleoside 5'-triphosphates and L-ATP, which are enantiomers of natural D-dNTPs and D-ATP, on deoxycytidine kinase (dCK) partially purified from mouse leukemic P388 cells were investigated. Only L-dCTP did not act as a phosphate donor while other L-dNTPs and L-ATP showed 15-30% of the activity of the corresponding D-dNTP or D-ATP. L-dCTP inhibited dCK non-competitively with 2'-deoxycytidine (D-dCyd) and competitively with phosphate donor D-ATP. These inhibitory effects of L-dCTP on dCK were similar to the results of earlier studies using D-dCTP. Thus, L-dCTP was shown to be capable of serving as a feedback inhibitor for dCK instead of D-dCTP. Mouse dCK was also able to phosphorylate L-dCyd, as demonstrated in the case of human dCK. The present results suggest that the chirality of not only dCyd as the substrate but also nucleotides as the substrate or effector is not strictly discriminated by dCK.
Asunto(s)
Desoxicitidina Quinasa/metabolismo , Desoxicitidina/metabolismo , Nucleótidos/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Citidina Trifosfato/metabolismo , Desoxicitidina/química , Nucleótidos de Desoxicitosina/metabolismo , Humanos , Ratones , Nucleótidos/química , Estereoisomerismo , Especificidad por SustratoRESUMEN
Photolabile 2'-deoxy- E -5-[4-(3-trifluoromethyl-3 H-diazirin-3-yl)styryl]uridine and its protected phosphoramidite derivatives have been synthesized and introduced into DNA oligomers through solid-phase DNA synthesis. The (trifluoromethyldiazirinyl)stylyl moiety of this nucleoside was found to be sufficiently stable for automated DNA synthesis. In addition, this moiety was found to be stable at 60 degrees C in aqueous solution under the annealing conditions for duplex formation with complementary strands, since >95% of the photolabile nucleoside remained after heating for 1 h. The oligo(dT) 15mer analog bearing the photolabile residue was activated/decomposed by near-UV irradiation. In photoaffinity cross-linking experiments with recombinant rat DNA polymerasebeta, constituted from a 40 kDa polypeptide, using oligo(dT) 15mer analogs bearing the photolabile residue near the 3'-terminus, a covalently bound complex of 45 kDa was obtained in the presence of complementary templates. Thus it was demonstrated that our method for synthesis of photolabile oligodeoxyribonucleotides may be useful for studies of DNA-related enzymes and DNA binding proteins.
Asunto(s)
Azirinas/síntesis química , ADN Polimerasa I/metabolismo , Oligodesoxirribonucleótidos/síntesis química , Sondas de Oligonucleótidos/síntesis química , Uridina/análogos & derivados , Animales , Azirinas/química , Secuencia de Bases , Reactivos de Enlaces Cruzados , Estabilidad de Medicamentos , Indicadores y Reactivos , Mamíferos , Oligodesoxirribonucleótidos/química , Sondas de Oligonucleótidos/química , Ratas , Proteínas Recombinantes/metabolismo , Espectrofotometría Ultravioleta , Termodinámica , Uridina/síntesis química , Uridina/químicaRESUMEN
2'-Deoxyuridine derivatives bearing an activated ester at C-5 position were synthesized and was examined their use for the preparation of modified oligodeoxyribonucleotides (ODNs) by a post-modification method. The ODNs containing cyanomethyl ester at C-5 position of the deoxyuridine residue reacted easily with a primary amine of several functional molecules under the mild condition to give the corresponding modified ODNs.