Selective Inhibition of HIV Replication by Adriamycin in Macrophages But Not in Lymphocytes

Bergamini, Alberto; Perno, Carlo Federico; Balzarini, Jan; Capozzi, M.; Marinelli, Leonardo; Milanese, G.; Pesce, Caterina Delfina; Caliò, R; Rocchi, G

doi:10.1089/aid.1992.8.1239

Cited by 12 publications

(6 citation statements)

References 19 publications

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“…Doxorubicin has also been shown to inhibit HIV replication in monocyte-macrophage cells, although this effect was limited to inhibition of initial infection rather than decreased viral replication in chronically infected cells (43). Therefore, it is likely that doxorubicin has some effect on the HIV-1 reverse-transcribed DNA integration into the cellular genome, as suggested by the authors (43), rather than having a downstream effect on the viral replication cycle at the doses tested. It should be noted, however, that since the levels of the protease, reverse transcriptase, and integrase proteins were not measured relative to the levels of p24 (capsid protein), a direct effect of doxorubicin on frameshifting in monocyte-macrophage cells has not been demonstrated.…”

Section: Resultsmentioning

confidence: 94%

Section: Resultsmentioning

confidence: 99%

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Selection and Characterization of Small Molecules That Bind the HIV-1 Frameshift Site RNA

2009

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HIV-1 requires a −1 translational frameshift to properly synthesize the viral enzymes required for replication. The frameshift mechanism is dependent upon two RNA elements, a seven-nucleotide slippery sequence (UUUUUUA) and a downstream RNA structure. Frameshifting occurs with a frequency of ~5%, and increasing or decreasing this frequency may result in a decrease in viral replication. Here, we report the results of a high-throughput screen designed to find small molecules that bind to the HIV-1 frameshift site RNA. Out of 34,500 compounds screened, 202 were identified as positive hits. We show that one of these compounds, doxorubicin, binds the HIV-1 RNA with low micromolar affinity (K d = 2.8 μM). This binding was confirmed and localized to the RNA using NMR. Further analysis revealed that this compound increased the RNA stability by approximately 5 °C and decreased translational frameshifting by 28% (±14%), as measured in vitro.Twenty-five years after its initial identification, human immunodeficiency virus type 1 (HIV-1) continues to be a major health concern across much of the world. Although many anti-HIV-1 drugs have been developed, the virus is often able to evade these therapies owing to its high mutation rate. In addition to new classes of drugs being designed against the traditional targets (protease and reverse transcriptase) (1,2), recent research has focused on targeting the −1 programmed translational frameshift that occurs between the gag and pol reading frames (3)(4)(5)(6)(7)(8). This frameshift allows for translation of the pol genes (encoding the protease, reverse transcriptase, and integrase enzymes) in the form of a Gag-Pol fusion polyprotein via the evasion of a stop codon located at the end of the gag gene. Without this frameshift, only the Gag polyprotein (matrix, capsid, nucleocapsid, and p6 structural proteins) will be produced. The structural and enzymatic proteins are found in approximately a 20:1 molar ratio as a result of a frameshift efficiency of approximately 5% (9-15). This stoichiometry is required for appropriate packaging of virus particles, and an increase or decrease in the frameshift efficiency has been found to significantly decrease the production of infectious virions (4,15,16).The frameshift event is programmed by two cis-acting RNA elements between the gag and pol genes: a seven-nucleotide slippery sequence (UUUUUUA) and a highly conserved stemloop structure immediately downstream (9,17). This stem-loop structure has been shown to induce ribosomal pausing, which is required for frameshifting, and its stability has been correlated with the efficiency of ribosomal frameshifting (18,19). At the base of the stem-loop, the frameshift site RNA contains a conserved GGA bulge. This sequence has been hypothesized to interact with the translational machinery during frameshifting (20), by virtue of its predicted positioning near the mRNA entrance channel, which is located 13-15 nucleotides of the P site codon (21). Due to the distance the mRNA must traverse through the r...

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Section: Resultsmentioning

confidence: 94%

Section: Resultsmentioning

confidence: 99%

Selection and Characterization of Small Molecules That Bind the HIV-1 Frameshift Site RNA

2009

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“…As described above, cytostatic and cytotoxic effects of test compounds in cell-based assays are often mistakenly interpreted as representative of apparent antiviral activity since the production of the relevant virus is reduced by the inherent cytostatic and cytotoxic effects of the test compounds (39,40). Indeed, none of such toxic agents (i.e., daunorubicin and adriamycin) have proven to be of clinical utility as antiviral agents.…”

Section: Discussionmentioning

confidence: 99%

GRL-0920, an Indole Chloropyridinyl Ester, Completely Blocks SARS-CoV-2 Infection

Hattori

Higshi-Kuwata

Raghavaiah

et al. 2020

mBio

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We assessed various newly generated compounds that target the main protease (Mpro) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and various previously known compounds reportedly active against SARS-CoV-2, employing RNA quantitative PCR (RNA-qPCR), cytopathicity assays, and immunocytochemistry. Here, we show that two indole-chloropyridinyl-ester derivatives, GRL-0820 and GRL-0920, exerted potent activity against SARS-CoV-2 in cell-based assays performed using VeroE6 cells and TMPRSS2-overexpressing VeroE6 cells. While GRL-0820 and the nucleotide analog remdesivir blocked SARS-CoV-2 infection, viral breakthrough occurred. No significant anti-SARS-CoV-2 activity was found for several compounds reportedly active against SARS-CoV-2 such as lopinavir, nelfinavir, nitazoxanide, favipiravir, and hydroxychroloquine. In contrast, GRL-0920 exerted potent activity against SARS-CoV-2 (50% effective concentration [EC50] = 2.8 μM) and dramatically reduced the infectivity, replication, and cytopathic effect of SARS-CoV-2 without significant toxicity as examined with immunocytochemistry. Structural modeling shows that indole and chloropyridinyl of the derivatives interact with two catalytic dyad residues of Mpro, Cys145 and His41, resulting in covalent bonding, which was verified using high-performance liquid chromatography–mass spectrometry (HPLC/MS), suggesting that the indole moiety is critical for the anti-SARS-CoV-2 activity of the derivatives. GRL-0920 might serve as a potential therapeutic for coronavirus disease 2019 (COVID-19) and might be optimized to generate more-potent anti-SARS-CoV-2 compounds. IMPORTANCE Targeting the main protease (Mpro) of SARS-CoV-2, we identified two indole-chloropyridinyl-ester derivatives, GRL-0820 and GRL-0920, active against SARS-CoV-2, employing RNA-qPCR and immunocytochemistry and show that the two compounds exerted potent activity against SARS-CoV-2. While GRL-0820 and remdesivir blocked SARS-CoV-2 infection, viral breakthrough occurred as examined with immunocytochemistry. In contrast, GRL-0920 completely blocked the infectivity and cytopathic effect of SARS-CoV-2 without significant toxicity. Structural modeling showed that indole and chloropyridinyl of the derivatives interacted with two catalytic dyad residues of Mpro, Cys145 and His41, resulting in covalent bonding, which was verified using HPLC/MS. The present data should shed light on the development of therapeutics for COVID-19, and optimization of GRL-0920 based on the present data is essential to develop more-potent anti-SARS-CoV-2 compounds for treating COVID-19.

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“…In the search for drugs to repurpose for COVID-19 disease, we suggest doxorubicin and paclitaxel be considered. These drugs have not been proposed hitherto although they have been evaluated for antiviral activity, particularly with respect to inhibition of viral helicase ( Ash and Diekema, 1987 ; Bergamini et al., 1992 ; Borowski et al., 2002 ; Briguglio et al., 2011 ). Paclitaxel also suppressed inflammation in a murine model of bacterial pneumonia ( Mirzapoiazova et al., 2007 ).…”

Section: Discussionmentioning

confidence: 99%

Vulnerabilities of the SARS-CoV-2 Virus to Proteotoxicity—Opportunity for Repurposed Chemotherapy of COVID-19 Infection

et al. 2020

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The global pandemic of COVID-19 disease caused by infection with the SARS-CoV-2 coronavirus, has produced an urgent requirement and search for improved treatments while effective vaccines are developed. A strategy for improved drug therapy is to increase levels of endogenous reactive metabolites for selective toxicity to SARS-CoV-2 by preferential damage to the viral proteome. Key reactive metabolites producing major quantitative damage to the proteome in physiological systems are: reactive oxygen species (ROS) and the reactive glycating agent methylglyoxal (MG); cysteine residues and arginine residues are their most susceptible targets, respectively. From sequenced-based prediction of the SARS-CoV-2 proteome, we found 0.8-fold enrichment or depletion of cysteine residues in functional domains of the viral proteome; whereas there was a 4.6-fold enrichment of arginine residues, suggesting SARS-CoV-2 is resistant to oxidative agents and sensitive to MG. For arginine residues of the SARS-CoV-2 coronavirus predicted to be in functional domains, we examined which are activated toward modification by MG – residues with predicted or expected low pK a by neighboring group in interactions. We found 25 such arginine residues, including 2 in the spike protein and 10 in the nucleoprotein. These sites were partially conserved in related coronaviridae: SARS-CoV and MERS. Finally, w e identified drugs which increase cellular MG concentration to virucidal levels: antitumor drugs with historical antiviral activity, doxorubicin and paclitaxel. Our findings provide evidence of potential vulnerability of SARS-CoV-2 to inactivation by MG and a scientific rationale for repurposing of doxorubicin and paclitaxel for treatment of COVID-19 disease, providing efficacy and adequate therapeutic index may be established.

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Selective Inhibition of HIV Replication by Adriamycin in Macrophages But Not in Lymphocytes

Cited by 12 publications

References 19 publications

Selection and Characterization of Small Molecules That Bind the HIV-1 Frameshift Site RNA

Selection and Characterization of Small Molecules That Bind the HIV-1 Frameshift Site RNA

GRL-0920, an Indole Chloropyridinyl Ester, Completely Blocks SARS-CoV-2 Infection

Vulnerabilities of the SARS-CoV-2 Virus to Proteotoxicity—Opportunity for Repurposed Chemotherapy of COVID-19 Infection

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