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Localization of calcitonin mRNA using in situ labeling with reverse transcriptase and PCR

100%

Zabel M. , Ciesielska U. , Wysocka T.

Folia Histochemica et Cytobiologica. Supplement

1996

tom 34

nr 1

Anti-HIV activities and mechanisms of antisense oligonucleotides

86%

Hatta T. , Inagawa T. , Kuwasaki T. , Kinzuka Y. , Takai K. , Yokoyama S. , Nakashima H. , Yamamoto N. , Takaku H.

Biotechnologia

1996

nr 4

Synthesis and anti-HIV properties of novel 6-phenylselenenyl-5-propyluracils

72%

Miazga A. , Felczak K. , Siwecka M. A. , Lipniacki A. , Piasek A. , Kulikowski T.

tom 54

nr 4

863-868

Novel 6-phenylselenenyl-5-propyluracils were synthesized from 5-propyluracil with the use of regioselective synthesis to give 1-[(2-hydroxyethoxy)-methyl]-6-phenylselenenyl-5-propyluracil (6), 1-ethoxymethyl-6-phenylselenenyl-5-propyluracil (9) and 1-benzyloxymethyl-6-phenylselenenyl-5-propyluracil (10). Interaction of these compounds with recombinant HIV-1 reverse transcriptase (RT) was evaluated using a non-isotopic colorimetric method. Compounds 9 and 10 exerted potent HIV RT inhibition (IC500.06 and 0.05 μM respectively) while compound 6 showed moderate inhibition (IC50=3.5 μM). Potent anti-HIV-1 activity in MT-2 cells inoculated by a syncythia-in-ducing HIV-1 (cat #3 strain) laboratory isolate was exerted by compounds 9 and 10 (EC500.62 μM and 0.025 μM, respectively), while compound 6 showed only moderate activity (IC50=4.1 μM). In addition, compound 10 showed very good in vitro therapeutic index (TI > 2046), indicating that it is a potential anti-HIV/AIDS drug.

Template-directed base pairing of 2-chloro-2'-deoxyadenosine catalyzed by AMV reverse transcriptase

72%

Bukowska-Maciejewska A. M. , Kusmierek J. T.

Acta Biochimica Polonica

1998

tom 45

nr 2

2-Chloro-2'-deoxyadenine (2CldA) is used for treatment of several lymphoid malignancies. Since this drug is incorporated into DNA, we have undertaken studies on base pairing of 2-chloroadenine (2ClA). 2CldA phosphoramidite was synthesized and used for preparation of 25-mer templates with 2ClA located at site 21 from the 3'-end. Kinetic parameters (Km and Vmax) for the incorporation of deoxynucleoside-5'-triphosphates by AMV reverse transcriptase opposite the 2ClA template, as well as for the extension of 2ClA·T pair, were determined. The efficiency (Vmax/Km) of incorporation of dGTP, dCTP, and dATP opposite 2ClA is at least one order of magnitude lower than opposite unmodified A. The efficiency of incorporation of dTTP opposite 2ClA is about 30-fold lower than opposite A and extension of 2ClA·T pair is 3-fold lower than of A·T pair. From the analysis of the parameters of dTTP incorporation we conclude that formation of 2ClA·T pair is thermodynamically, but not kinetically controlled. The difference in binding energy (ΔΔG) between 2ClA·T and A·T pairs in the environment of the polymerase active site is 2 kcal/mol. Our results indicate that the presence of 2ClA in DNA slows down replication, but does not lead to base-substitution mutations.