Inhibition of the Cellular Rev Response and HIV-1 Replication by 8-Alkyl-2-(4-pyridyl)pyrido[2,3-<i>d</i> pyrimidin-5(8<i>H</i>)-Ones

Ciccarelli, Richard B.; Winter, Laurie A.; Lorenz, Roman R.; Harris, A.; Crawford, Ann; Bailey, T. R.; Singh, Balraj; Hammarskjöld, Marie‐Louise; Rekosh, David; Hughes, Joseph V.

doi:10.1177/095632029400500305

Cited by 7 publications

(3 citation statements)

References 10 publications

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“…In contrast, our previous efforts in this area used a cell-based screen that involved transient transfection with expression vec- tors producing Rev and Rev-dependent HIV GagPol (10). p24 secretion into the medium was used as an end point to measure possible inhibition of Rev function.…”

Section: Discussionmentioning

confidence: 99%

Heterocyclic Compounds That Inhibit Rev-RRE Function and Human Immunodeficiency Virus Type 1 Replication

Shuck-Lee

Chen

Willard

et al. 2008

Antimicrob Agents Chemother

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A cell-based screening assay was performed to identify compounds that inhibited the postintegration stage of the human immunodeficiency virus (HIV) life cycle. This assay utilized a cell line that contains the HIV gag and pol genes expressed in a Rev-dependent fashion. The cell line produces about 10 to 15 ng of p24 per milliliter of medium over a 24-h period in the form of viruslike particles. Any compound that inhibits a postintegration step in the HIV life cycle scores in this assay by decreasing particle production. Forty thousand compounds were screened, and 192 compounds were selected from the original screen because they showed more than 50% inhibition at a 10 M concentration. The cumulative evidence presented in this study strongly suggests that 2 of the 192 compounds work as inhibitors of HIV Rev function. This was determined by a variety of cell-based assays, although the compounds do not interfere with Rev-RRE (Rev response element) binding in vitro. Both compounds inhibit replication of the lab isolate NL4-3 as well as an HIV primary isolate from Brazil (93BR021) and thus are promising leads as therapeutic candidates that target HIV replication through inhibition of Rev function.Most of the current drugs in use for the treatment of AIDS work by targeting the enzymatic activities of the human immunodeficiency virus (HIV) reverse transcriptase or protease, although entry (7) and integrase (13) inhibitors are starting to be used, and presently there is also promising development of other novel targets (51, 59). However, because of the emergence of drug-resistant virus that commonly occurs as the result of treatment, there remains a great need to continue the search for alternative therapies that target other essential viral activities.The Rev protein is absolutely essential for HIV replication (for a review see reference 49). Proviral clones lacking a functional rev gene have no replicative ability, even in established tissue culture cell lines or peripheral blood mononuclear cells (PBMCs). In the absence of Rev, genomic RNA and several other HIV mRNAs cannot exit the nucleus (22,30,42). Thus, viral structural proteins are not made and the infectious cycle cannot continue. It is thus clear that modalities inhibiting the function of Rev could form the basis for therapy against HIV infection and AIDS.Although the Rev/RRE (Rev response element) export pathway is still not fully understood, several important steps have been identified (see reference 49). The pathway starts with the import of Rev into the nucleus (34, 58). Rev then binds specifically to RNA containing the RRE (17,28,30,42,53) and multimerizes on the RRE in a process believed to involve protein-protein as well as protein-RNA interactions (12,14,16,32,36,43,67). The Rev-RRE complex is then recognized by Crm1 (exportin 1; official gene symbol, XPO1) and RAN-GTP (1), which initiates the export process and eventually targets the complex to the nuclear pore, where it interacts with nucleoporins (1,4,25,70). This results in translocation of the com...

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Section: Discussionmentioning

confidence: 99%

Heterocyclic Compounds That Inhibit Rev-RRE Function and Human Immunodeficiency Virus Type 1 Replication

Shuck-Lee

Chen

Willard

et al. 2008

Antimicrob Agents Chemother

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“…A small-molecule inhibitor of Rev was discovered at Sterling-Winthrop (now Sanofi-Winthrop), using a 96-well plate assay to measure Rev function in transfected cells (108). The assay measured production of the p24 protein from the HIV gag-pol gene as a result of Rev expression, in COS-1 cells.…”

Section: Screening Approachesmentioning

confidence: 99%

“…At Oncogene Science, Inc., we are carrying out high throughput screening seeking Rev inhibitors, using a cell-based assay similar to the one used at Sterling-Winthrop (108). This program is in the early phases, and we hope to contribute new entities with new activities.…”

Section: Screening Approachesmentioning

confidence: 99%

Inhibition of the HIV Rev transactivator A new target for therapeutic intervention

Heguy¹

1997

Front Biosci

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The viral transactivator Rev is essential for HIV replication, since it allows the nuclear export of unspliced and partially spliced viral mRNAs that encode the structural proteins. Rev is an RNA binding protein that interacts with a highly structured RNA element, the RRE, found within the envelope sequences. This viral protein also interacts with cellular proteins, termed nucleoporins, and acts as an adaptor between the viral mRNAs and the cellular nuclear export machinery. Both interactions are specific, and required for Rev function. Because of its crucial role in the HIV replication cycle, and its novel mechanism of action, Rev represents an ideal target for therapeutic intervention. This review describes the efforts towards Rev inhibition. Gene therapy approaches, including the expression of trans-dominant mutants and RNA decoys, as well as antisense therapies and small molecule inhibitors of Rev-RRE binding or Rev interaction with the cellular machinery will be discussed. INTRODUCTIONHIV-1, the etiologic agent of Acquired Immunodeficiency Syndrome (AIDS), displays a complex regulation of __________________________________________ Received 6/2/97 Accepted 6/4/97 1 To whom correspondence should be addressed at: Oncogene Science, Inc., 106 Charles Lindbergh Blvd., Uniondale, NY 11553 Tel: 516-222-0023 Fax: 516-222-0114 E-mail: aheguy@oncogene.com viral gene expression during its life cycle. Unlike many "simple" retroviruses (i.e. avian and murine leukemia viruses), which express only three viral genes, the genome of HIV-1 encodes nine genes whose expression patterns are tightly regulated during the HIV-1 replication cycle (Figure 1, for reviews see 1, 2, 3). In the infected host cell, HIV expresses over 20 distinct mRNA species (reviewed in 4). The early stage of regulation of the HIV-1 life cycle is marked by the appearance of the viral regulatory molecules Tat, Rev, and Nef, encoded by the fully spliced 2 kb class of viral mRNAs. The late viral life cycle gene expression is characterized by the cytoplasmic appearance of the 4 kb class of single spliced and 9.2 kb unspliced mRNAs, that encode the proteins required for the assembly of infectious virions. The viral transactivator Rev allows this transition into the late cycle (5, 6), and is therefore essential for viral replication. In effect, proviral mutants that do not express Rev fail to produce structural proteins and therefore cannot form new infectious viral particles (5, 7).Because of its essential role in HIV replication, Rev constitutes an excellent target for therapeutic intervention. Its mode of action and specific interactions with its target RNA and cellular proteins have been extensively studied and elegantly elucidated, and this body of knowledge adds to the attractiveness of Rev as a target. The purpose of this article is to briefly review the latest developments on Rev, and how this knowledge can be used for development of anti-viral strategies, as well as to

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Practical high throughput screening

Reichman

Harris²

1997

Annual Reports in Combinatorial Chemistry and Molecular Diversity

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Inhibition of the Cellular Rev Response and HIV-1 Replication by 8-Alkyl-2-(4-pyridyl)pyrido[2,3-d pyrimidin-5(8H)-Ones

Cited by 7 publications

References 10 publications

Heterocyclic Compounds That Inhibit Rev-RRE Function and Human Immunodeficiency Virus Type 1 Replication

Heterocyclic Compounds That Inhibit Rev-RRE Function and Human Immunodeficiency Virus Type 1 Replication

Inhibition of the HIV Rev transactivator A new target for therapeutic intervention

Practical high throughput screening

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