RESUMEN
A tin(II) complex coordinated by a sterically demanding o-phenylenediamido ligand is synthesized. The ligand is redox-active to reach a tin(II) complex with the diiminobenzosemiquinone radial anion in the oxidation by AgPF6. The tin(II) complex reacts with a series of nosylazides (x-NO2C6H4-SO2-N3; x = o, m, or p) at -30 °C to yield the corresponding nitrene radical bound tin(II) complexes. The nitrene radical complexes exhibit C(sp3)-H activation and amination reactivity.
RESUMEN
Oligodeoxynucleotides (ODN) containing CpG motifs (CpG) act as modulators that bias the immune response towards a Th1-dominant phenotype. To investigate this effect further, we examined the protective effects of a covalently linked conjugate between CpG-ODN and HA-2kd antigen in mice infected with influenza A virus. The conjugated form of CpG-ODN and HA-2kd was more efficient in regulating influenza A virus than the unconjugated mixture of CpG-ODN and HA-2kd. The antigen-conjugated CpG-ODN induced an immune response with a Th1-dominant cytokine pattern characterized by the secretion of high levels of HA-2kd-specific interferon-gamma and IgG2a (Th1), which were only slightly induced by HA-2kd alone. These findings support the use of CpG-ODN-Ag conjugates as novel Ag-specific immunomodulators and suggest that CpG-ODN-HA-2kd might be a promising immune therapy for patients with influenza virus.
Asunto(s)
Antivirales/farmacología , Islas de CpG , Inmunoterapia/métodos , Virus de la Influenza A/metabolismo , Oligodesoxirribonucleótidos/farmacología , Infecciones por Orthomyxoviridae/prevención & control , Animales , Antígenos/química , Antígenos CD8/biosíntesis , Línea Celular , Citocinas/metabolismo , Perros , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo , Inmunoglobulina G/química , Inflamación , Interferón gamma/metabolismo , Ratones , Ratones Endogámicos BALB C , Oligodesoxirribonucleótidos/química , Oligonucleótidos/farmacología , Péptidos/química , Fenotipo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Células TH1/inmunología , Células TH1/virología , Factores de TiempoRESUMEN
Oligodeoxynucleotides with RNA cleavage activity 1) were conjugated with amines and peptides by solid phase fragment condensation (SPFC). It was found that 29 mer DNA enzyme conjugated with spermine at its 5'-end showed higher affinity to the target RNA sequence and 40 times higher activity of cleavage than native DNA enzyme. It is also to be noted that conjugate DNA enzymes showed increased resistance against nuclease digestion.
Asunto(s)
ADN/química , Oligodesoxirribonucleótidos/química , Secuencia de Bases , Catálisis , Hibridación de Ácido Nucleico , EsperminaAsunto(s)
Proteínas Nucleares , Oligonucleótidos/síntesis química , Factores de Transcripción , Secuencia de Aminoácidos , Proteína con Homeodominio Antennapedia , ADN sin Sentido/síntesis química , ADN sin Sentido/química , Proteínas de Drosophila/química , Proteínas de Drosophila/genética , Productos del Gen tat/química , Productos del Gen tat/genética , Proteínas de Homeodominio/química , Proteínas de Homeodominio/genética , Hidróxidos , Indicadores y Reactivos , Datos de Secuencia MolecularRESUMEN
Development of a novel method for the synthesis of DNA conjugates is described. Oligonucleotides were successfully conjugated with a variety of functional molecules on a solid phase (Solid Phase Fragment Condensation) using an amino, a hydroxyl, a thiol, and a carboxyl group. DNA-peptide conjugate was obtained as a pure from by a single RPHPLC purification approximately in 20% yield. Moreover, it was demonstrated that the present method was effective for the preparation of conjugate molecules, DNA-sugar, DNA-polyamine, DNA-lipid and so on. The study to create new intelligent DNAs by accumulation various biofunctions on the molecule by SPFC is now in progress in our laboratory.
Asunto(s)
ADN/síntesis química , Secuencia de Bases , ADN/química , Indicadores y Reactivos , Oligodesoxirribonucleótidos/química , Fragmentos de Péptidos/químicaAsunto(s)
ADN Catalítico/metabolismo , ADN/metabolismo , ARN/metabolismo , Ribonucleasas/metabolismo , Secuencia de Bases , Sitios de Unión , ADN/química , Estabilidad de Enzimas , Ingeniería Genética , Oligodesoxirribonucleótidos/química , Oligorribonucleótidos/química , ARN/química , ARN Catalítico/metabolismo , Especificidad por SustratoRESUMEN
Various types of DNA-peptide conjugates were synthesized by solid phase fragment condensation (SPFC). DNA-LNS (nuclear localizing signal) peptide conjugate was proved to be delivered and localized into cellular nucleus and exhibited higher antisense inhibitory effect against telomerase than antisense phosphorothioate DNA. In contrast, DNAzymes conjugated with NES (nuclear export signal) peptide was shown to be taken up and localized in cytoplasm. Inhibitory effect of the conjugate DNAzyme against BCR-ABL tyrosine kinase was evaluated to be more significant than the native DNAzyme.
Asunto(s)
ADN/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Péptidos/farmacología , Secuencia de Aminoácidos , ADN/química , Humanos , Células Jurkat , Péptidos/químicaRESUMEN
CpG oligonucleotides have recently been highlighted as an immunomodulator that biases toward a Th1-dominant phenotype. To substantiate the synergism between CpG-ODNs and Ag, we introduced a linked conjugate between CpG-ODNs and Ag and the antiviral effect of antigen-conjugated CpG-ODNs therapy following an infection of mice with the influenza virus A. We found that the conjugate inhibited the influenza virus infection more efficiently than the coadministration of the unconjugated CpG-ODNs and the Ag mixture. CpG-ODNs is effective in immunity induction and will be useful in treating virus infections.
