       Document 0436
 DOCN  M9640436
 TI    Nuclear localization signal of HIV-1 as a novel target for therapeutic
       intervention.
 DT    9604
 AU    Dubrovsky L; Ulrich P; Nuovo GJ; Manogue KR; Cerami A; Bukrinsky M;
       Picower Institute for Medical Research, Manhasset, New York; 11030, USA.
 SO    Mol Med. 1995 Jan;1(2):217-30. Unique Identifier : AIDSLINE MED/96091345
 AB    BACKGROUND: Human immunodeficiency virus type 1 (HIV-1) is a lentivirus
       and shares with other members of this retroviral subfamily the ability
       to replicate in nondividing cells, in particular, cells of the
       monocyte/macrophage lineage. This feature relies on the presence of a
       specific nuclear localization signal (NLS) within the viral matrix
       protein (MA p17), which to some degree can be complemented by the
       activity of the viral vpr gene product. The MA p17 NLS ensures efficient
       transportation of the viral preintegration complex into the nucleus of
       an infected macrophage and confers persistence of HIV-1 in quiescent T
       cells, and therefore presents an attractive target for therapeutic
       intervention. MATERIALS AND METHODS: Nuclear localization signals (NLS)
       in general and the HIV-1 MA p17 NLS in particular are characterized by a
       stretch of positively charged amino acids including one or more lysine
       residues. A series of compounds potentially capable of binding and
       reacting with lysine by forming Schiff base adducts was synthesized. Our
       special consideration was to make compounds that would preferentially
       bind to two closely contiguous amino functions, as opposed to isolated
       single lysine residues. We assumed that this approach might specifically
       target the compound to NLS while affecting other regions less, thus
       reducing nonspecific cytotoxicity. Antiviral activity was assessed in
       primary monocytes and in peripheral blood lymphocytes (PBL) infected
       with HIV-1ADA strain. Viral replication was monitored by reverse
       transcriptase (RT) activity in the supernatant. Efficiency of nuclear
       importation of the viral preintegration complex was estimated by the
       formation of 2-LTR circle forms of HIV-1 DNA and also by in situ PCR
       techniques. RESULTS: Arylene bis(methyl ketone) compounds with a
       nitrogenous third subsituent, especially a pyrimidinic side-chain,
       inhibited HIV-1 replication in human monocytes at an IC50 as low as 1
       nM. These compounds did not block HIV-1 replication in peripheral blood
       lymphocyte cultures. The inhibitory effect observed in monocyte cultures
       appeared in the context of markedly reduced nuclear importation of viral
       DNA in the presence of the drug. No cytotoxic effects of the compounds
       was observed in vitro at concentrations as high as 10 microM. An
       amidinohydrazone derivative of the most active compound was about 100
       times less active than the parent, indicating that carbonyl groups were
       instrumental in the antiviral effect. CONCLUSIONS: These early results
       suggest that retroviral replication in nondividing cells is susceptible
       to pharmaceutical intervention targeted against the NLS activity of
       HIV-1 proteins in the viral preintegration complex. The compounds
       described efficiently block translocation of viral DNA to the nuclei of
       infected primary monocytes, and inhibit viral replication. This
       inhibition is effective only in nondividing cells and is not seen in
       proliferating cultures, such as activated PBLs. Thus, drugs that target
       HIV-1 NLS may be useful to specifically block the macrophage arm of HIV
       infection and could thereby be of value in treating macrophage-specific
       manifestations of HIV disease, such as HIV-1 dementia. In combination
       with other drugs, potential therapeutics exploiting this target may also
       help to control the progression of HIV-1 infection and disease.
 DE    Antiviral Agents/CHEMICAL SYNTHESIS/*PHARMACOLOGY  Cell
       Nucleus/*VIROLOGY  Cells, Cultured  Dose-Response Relationship, Drug
       Human  HIV Core Protein p24/ANALYSIS  HIV Long Terminal Repeat
       HIV-1/DRUG EFFECTS/GENETICS/*PHYSIOLOGY  Lymphocytes/*VIROLOGY
       Monocytes/*VIROLOGY  Pyrimidines/CHEMICAL SYNTHESIS/*PHARMACOLOGY
       RNA-Directed DNA Polymerase/ANALYSIS  Support, Non-U.S. Gov't  Support,
       U.S. Gov't, P.H.S.  *Virus Replication/DRUG EFFECTS  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

