Kinetic studies with the non-nucleoside HIV-1 reverse transcriptase inhibitor U-88204E.

The bis(heteroaryl)piperazine U-88204E is a potent inhibitor of HIV-1 reverse transcriptase (RT) and possesses excellent anti-HIV activity in HIV-1-infected lymphocytes grown in tissue culture. Enzymatic kinetic studies of the RNA- and DNA-dependent DNA polymerases of RT were carried out in order to determine whether the inhibitor interacts directly with the template:primer or deoxyribonucleotide triphosphate (dNTP) binding sites of the polymerase. The experimental results were analyzed using steady-state or Briggs-Haldane kinetics, by assuming that the template:primer binds to the enzyme first followed by the dNTP and that the polymerase functions processively. The results of the analysis show that the inhibitor acts as a mixed to noncompetitive inhibitor with respect to both the template:primer and the dNTP binding sites. The potency of U-88204E on the RNA-directed DNA polymerase activity depends on the base composition of the template:primer. The Ki values for the poly(rC):(dG)10-directed reactions were at least 7 times lower than the ones for reactions directed by poly(rA):(dT)10. The inhibitor did not inhibit the RNase H function of HIV-1 RT nor did it impair the RNA-directed DNA polymerase activity of HIV-2 RT. These data thus demonstrate the unique specificity of U-88204E for HIV-1 RT.

[1]  J. Chou,et al.  Steady-state kinetic studies with the non-nucleoside HIV-1 reverse transcriptase inhibitor U-87201E. , 1993, The Journal of biological chemistry.

[2]  J. Coleman,et al.  Conformational changes of HIV reverse transcriptase subunits on formation of the heterodimer: correlation with kcat and Km. , 1992, Biochemistry.

[3]  J. Reardon.  Human immunodeficiency virus reverse transcriptase: steady-state and pre-steady-state kinetics of nucleotide incorporation. , 1992, Biochemistry.

[4]  D. Chattopadhyay,et al.  Resolution of microheterogeneity associated with recombinant HIV-1 heterodimeric reverse transcriptase. , 1992, Protein expression and purification.

[5]  Cheng Yung-Chi,et al.  HIV-1 reverse transcriptase inhibition by a dipyridodiazepinone derivative: BI-RG-587. , 1992 .

[6]  T. J. Dueweke,et al.  The binding of a novel bisheteroarylpiperazine mediates inhibition of human immunodeficiency virus type 1 reverse transcriptase. , 1992, The Journal of biological chemistry.

[7]  L. Resnick,et al.  Nonnucleoside reverse transcriptase inhibitors that potently and specifically block human immunodeficiency virus type 1 replication. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[8]  W. Schleif,et al.  Pyridinone derivatives: specific human immunodeficiency virus type 1 reverse transcriptase inhibitors with antiviral activity. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[9]  E. De Clercq,et al.  Potent and selective inhibition of human immunodeficiency virus type 1 (HIV-1) by 5-ethyl-6-phenylthiouracil derivatives through their interaction with the HIV-1 reverse transcriptase. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[10]  C. Tan,et al.  Functional characterization of RNA-dependent DNA polymerase and RNase H activities of a recombinant HIV reverse transcriptase. , 1991, Biochemistry.

[11]  J. Proudfoot,et al.  A novel dipyridodiazepinone inhibitor of HIV-1 reverse transcriptase acts through a nonsubstrate binding site. , 1991, Biochemistry.

[12]  E. De Clercq,et al.  An antiviral target on reverse transcriptase of human immunodeficiency virus type 1 revealed by tetrahydroimidazo-[4,5,1-jk] [1,4]benzodiazepin-2 (1H)-one and -thione derivatives. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[13]  R. Ruprecht,et al.  N-carboxymethylchitosan-N,O-sulfate as an anti-HIV-1 agent. , 1991, Biochemical and biophysical research communications.

[14]  M. Baba,et al.  Potential anti-AIDS agents. Synthesis and antiviral activity of naphthalenesulfonic acid derivatives against HIV-1 and HIV-2. , 1991, Journal of medicinal chemistry.

[15]  J. Adams,et al.  Inhibition of HIV-1 replication by a nonnucleoside reverse transcriptase inhibitor. , 1990, Science.

[16]  K. Neet,et al.  Demonstration of a slow conformational change in liver glucokinase by fluorescence spectroscopy. , 1990, The Journal of biological chemistry.

[17]  C. Merril,et al.  Sodium pentosan polysulfate (PPS), an anti-HIV agent also exhibits synergism with AZT, lymphoproliferative activity, and virus enhancement. , 1990, AIDS research and human retroviruses.

[18]  E. De Clercq,et al.  Dextran sulfate and other polyanionic anti-HIV compounds specifically interact with the viral gp120 glycoprotein expressed by T-cells persistently infected with HIV-1. , 1990, Virology.

[19]  M. Deibel,et al.  Denaturation/refolding of purified recombinant HIV reverse transcriptase yields monomeric enzyme with high enzymatic activity. , 1990, AIDS research and human retroviruses.

[20]  Erik De Clercq,et al.  Potent and selective inhibition of HIV-1 replication in vitro by a novel series of TIBO derivatives , 1990, Nature.

[21]  K. Chou.  Applications of graph theory to enzyme kinetics and protein folding kinetics. Steady and non-steady-state systems. , 2020, Biophysical chemistry.

[22]  R T Walker,et al.  Highly specific inhibition of human immunodeficiency virus type 1 by a novel 6-substituted acyclouridine derivative. , 1989, Biochemical and biophysical research communications.

[23]  K. Moelling,et al.  Inhibition of human immunodeficiency virus type 1 RNase H by sulfated polyanions , 1989, Journal of virology.

[24]  K. Chou.  Graphic rules in steady and non-steady state enzyme kinetics. , 1989, The Journal of biological chemistry.

[25]  L. Resnick,et al.  Nucleotide dimers suppress HIV expression in vitro. , 1988, AIDS research and human retroviruses.

[26]  E. De Clercq,et al.  Sulfated polysaccharides are potent and selective inhibitors of various enveloped viruses, including herpes simplex virus, cytomegalovirus, vesicular stomatitis virus, and human immunodeficiency virus , 1988, Antimicrobial Agents and Chemotherapy.

[27]  Samuel H. Wilson,et al.  Studies on the mechanism of human immunodeficiency virus reverse transcriptase. Steady-state kinetics, processivity, and polynucleotide inhibition. , 1988, The Journal of biological chemistry.

[28]  E. De Clercq,et al.  Pentosan polysulfate, a sulfated oligosaccharide, is a potent and selective anti-HIV agent in vitro. , 1988, Antiviral research.

[29]  E. Clercq.  Suramin in the treatment of AIDS: mechanism of action. , 1987 .

[30]  K. Chou,et al.  Role of the protein outside active site on the diffusion-controlled reaction of enzymes , 1982 .

[31]  C Kuo-Chen.  The kinetics of the combination reaction between enzyme and substrate. , 1976, Scientia Sinica.