Template-competitive inhibitors of HIV-1 reverse transcriptase

Design, synthesis and inhibitory activity

Ke Li, Weiying Lin, Kar Hua Chong, Bob Moore, Michael B. Doughty

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We report the design, synthesis and activity studies on a novel class of template-competitive reverse transcriptase inhibitors (TCRTIs). The TCRTIs are 1,N6-etheno analogues of a series of dATP-based template-competitive DNA polymerase inhibitors synthesized in our laboratory (Moore, B. M.; Jalluri, R.; Doughty, M.B. Biochemistry 1996, 35, 11634). Thus, nucleotides 2-(4-azidophenacyl)thio-1,N6-etheno-2′-deoxyadenosine 5′-triphosphate 1, the tetrafluoro analogue 2-(4-azido-2,3,5,6-tetrafluorophenacyl)thio-1,N6-etheno-2′- deoxyadenosine 5′-triphosphate 2 and its analogues were synthesized by alkylation of 2-thio-1,N6-etheno-2′-deoxyadenosine 5′-monophosphate with the corresponding chloro- or bromo-alkyl halides and converted to the triphosphate. Kinetically, nucleotides 1 and 2 are both competitive inhibitors of reverse transcriptase versus template/primer with K(i)'s of 8.0 and 7.4 μM, respectively, and non-competitive inhibitors versus TTP with K(i)'s of 15 and 10 μM, respectively. Nucleotide 3, which differs from 1 only in that it lacks the etheno group, non-complementary nucleotide triphosphates, and related monophosphates and nucleosides, are completely inactive as inhibitors of reverse transcriptase at concentrations up to 1 mM. Photoinactivation of RT by 1 was both time- and concentration-dependent, and protected by template/primer but not by dNTPs. The concentration-dependent inactivation data gave a KD,app of 17.2 μM and maximum inactivation of 90%, and radiolabeled [β,γ-32P]-1 photoincorporated specifically and covalently into the p66 subunit of RT. Thus the photoinactivation data support our main conclusion from the kinetic data that this class of RT inhibitors are non-substrate and template-competitive.

Original languageEnglish (US)
Pages (from-to)507-515
Number of pages9
JournalBioorganic and Medicinal Chemistry
Volume10
Issue number3
DOIs
StatePublished - Feb 11 2002

Fingerprint

Reverse Transcriptase Inhibitors
Thermodynamic properties
Nucleotides
Nucleic Acid Synthesis Inhibitors
Biochemistry
Alkylation
Nucleosides
Application programs
Kinetics
Human immunodeficiency virus 1 reverse transcriptase
1,N(6)-ethenodeoxyadenosine
triphosphoric acid
2'-deoxyadenosine triphosphate

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

Template-competitive inhibitors of HIV-1 reverse transcriptase : Design, synthesis and inhibitory activity. / Li, Ke; Lin, Weiying; Chong, Kar Hua; Moore, Bob; Doughty, Michael B.

In: Bioorganic and Medicinal Chemistry, Vol. 10, No. 3, 11.02.2002, p. 507-515.

Research output: Contribution to journalArticle

Li, Ke ; Lin, Weiying ; Chong, Kar Hua ; Moore, Bob ; Doughty, Michael B. / Template-competitive inhibitors of HIV-1 reverse transcriptase : Design, synthesis and inhibitory activity. In: Bioorganic and Medicinal Chemistry. 2002 ; Vol. 10, No. 3. pp. 507-515.
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