RESUMO
The solution-phase, parallel synthesis and evaluation of a library of 132 (+)-1,2,9,9a-tetrahydrocyclopropa[c]benz[e]indol-4-one (CBI) analogues of CC-1065 and the duocarmycins containing dimeric monocyclic, bicyclic, and tricyclic heteroaromatic replacements for the DNA-binding domain are described. This systematic study revealed clear trends in the structural requirements for observation of potent cytotoxic activity and DNA alkylation efficiency, the range of which spans a magnitude of > or =10 000-fold. Combined with related studies, these results highlight that the role of the DNA-binding domain goes beyond simply providing DNA-binding selectivity and affinity (10-100-fold enhancement in properties), consistent with the proposal that it contributes significantly to catalysis of the DNA alkylation reaction accounting for as much as an additional 1000-fold enhancement in properties.
Assuntos
Antibióticos Antineoplásicos/síntese química , Antibióticos Antineoplásicos/farmacologia , Leucomicinas/síntese química , Alquilantes/síntese química , Alquilantes/química , Alquilantes/farmacologia , Alquilação/efeitos dos fármacos , Antibióticos Antineoplásicos/química , Sítios de Ligação , Técnicas de Química Combinatória , Ciclopropanos/química , DNA/metabolismo , DNA Viral/efeitos dos fármacos , DNA Viral/metabolismo , Duocarmicinas , Indóis/química , Concentração Inibidora 50 , Leucomicinas/química , Leucomicinas/farmacologia , Pirróis/síntese química , Pirróis/química , Pirróis/farmacologia , Pirrolidinonas/síntese química , Pirrolidinonas/química , Pirrolidinonas/farmacologia , Vírus 40 dos Símios/genética , Relação Estrutura-AtividadeRESUMO
The sequencing of DNA by current procedures involves the use of radioisotopic or fluorescent labels. We propose that stable isotopes can be used as such labels and that the large number of stable isotopes available would allow multiplexing so that many DNA segments could be sequenced simultaneously. We have developed methods to use 57Fe2O3 to synthesize ferrocene and to attach the ferrocene to the 5' end of oligonucleotides. The 57Fe-labeled M13 universal primer functioned normally in a Sanger sequencing procedure. When a 57Fe-labeled oligonucleotide had migrated on a polyacrylamide gel it was readily located on the dried gel by scanning with resonance ionization spectroscopy (RIS) coupled with mass spectrometry. Using a 57Fe-labeled primer in a PCR reaction a 2000-bp DNA was produced that was detected by RIS on nylon membrane after agarose electrophoresis. The rapid analysis features of RIS coupled with the multispectral multiplexing possibilities of stable isotopes should significantly increase the rate of determination of DNA sequences.