Dehydroepiandrosterone, epiandrosterone and synthetic derivatives inhibit Junin virus replication in vitro

doi:10.1016/j.virusres.2008.03.014

Virus Research

Volume 135, Issue 2, August 2008, Pages 203-212

https://doi.org/10.1016/j.virusres.2008.03.014 Get rights and content

Abstract

In the present paper the in vitro antiviral activity of dehydroepiandrosterone (DHEA), epiandrosterone (EA) and 16 synthetic derivatives against Junin virus (JUNV) replication in Vero cells was studied. DHEA and EA caused a selective inhibition of the replication of JUNV and other members of the Arenaviridae family such as Pichinde virus and Tacaribe virus. The compounds were not virucidal to cell-free JUNV. The impairment of viral replication was not due to an inhibitory effect of the steroids on virus adsorption or internalization. An inhibitory effect of the compounds on JUNV protein synthesis and both intracellular and extracellular virus production was demonstrated. A partial inhibitory action on cell surface expression of JUNV glycoprotein G1 was also detected on DHEA- and EA-treated cultures. Like DHEA and EA, three compounds obtained from EA by chemical synthesis showed selectivity indexes higher than ribavirin, the only antiviral compound that has shown partial efficacy against arenavirus infections.

Introduction

Arenaviruses are enveloped, single stranded, ambisense RNA viruses with a segmented genome consisting of two segments, designated large (L) and small (S). Most arenaviruses induce a persistent infection in their rodent reservoirs, and humans may be accidental hosts who become infected by contact with the carrier rodents or their excreta. Among the 23 arenavirus species known, 5 are associated with viral hemorrhagic fevers: Lassa, Junin, Machupo, Guanarito and Sabia. The danger of pathogenic arenaviruses for human health, linked to their increased emergence in recent years, and the lack of a totally effective chemotherapy for treatment, support their inclusion in the Category A Pathogen List of the Centers for Disease Control and Prevention (CDC) as potential agents of bioterrorism (Damonte and García, 2005).

Junin virus (JUNV) is the etiological agent of Argentine hemorrhagic fever (AHF), an endemo-epidemic disease that affects rural workers of the most fertile areas of the country. The administration of defined doses of convalescent plasma is up today the best therapy against AHF whereas ribavirin is the preferred method of treatment for Lassa fever; however, undesirable side effects were recorded (McCormick et al., 1986, Enria and Maiztegui, 1994).

Previous reports showed that steroids inhibit in vitro replication of the arenaviruses JUNV, Tacaribe virus (TCRV) and Pichinde virus (PICV), but their mechanisms of action are largely yet to be established (Wachsman et al., 2000, Wachsman et al., 2004, Garrido Santos et al., 2003, Castilla et al., 2005). These previous studies prompted us to further investigate the potential anti-arenavirus activity of other steroidal compounds.

Dehydroepiandrosterone (DHEA) and its sulfate ester (DHEA-S) are the most abundant circulating steroid hormones in humans. A number of potentially beneficial effects, such as anti-aging, anti-inflammatory, immunomodulatory, antiviral, anticancer and neurotropic effects, have been attributed to replacement therapy with these compounds based on findings in rodent models, in vitro studies and human populations (Nawata et al., 2002, Labrie et al., 2003, Yoshida et al., 2003). Studies performed to understand the molecular basis of these multi-functional properties led to identify genomic and rapid non-genomic mechanisms of DHEA action. In contrast to other steroid hormones a cell membrane receptor for DHEA has been proposed and DHEA binding to this receptor would trigger non-genomic effects by modulating mitogen-activated protein kinase (MAPK) and the serine/threonine kinase (Akt) cell singnaling pathways (Liu et al., 2007).

Because DHEA is a native steroid that has been used clinically with minimal side effects, its possible use as an antiviral agent against arenaviruses deserves study. However, DHEA administration for extended periods increases circulating testosterone and dihydrotestosterone manifold above normal levels, especially in women, and may cause masculinization (Labrie et al., 2003). Therefore, the evaluation of the antiviral activity of structural related compounds would also be useful in the search of an effective treatment for arenavirus infections. In the present study we examined the ability of DHEA, epiandrosterone (EA) (a DHEA metabolite that lacks of androgenic activity) and 16 synthetic derivatives obtained from DHEA or EA, to selectively inhibit arenavirus replication in cell cultures.

Section snippets

Compounds

DHEA, EA and ribavirin were purchased from Sigma–Aldrich Chemical Co. The compounds D1 to D4 (obtained from DHEA) and E1 to E12 (obtained from EA) were prepared in our laboratory. They were purified by flash chromatography on silica and identified by electron impact mass spectra (EI-MS), IR and ¹H and ¹³C NMR spectroscopy. EI-MS were measured in a VG Trio-2 or in a Shimadzu QP-5000 mass spectrometer at 70 eV by direct inlet. IR spectra were recorded in KBr pellets on a Nicolet Magna IR 550 FT

Antiviral activity of DHEA and EA against JUNV, PICV and TCRV

First we assessed cell viability of Vero cell cultures treated for 24 h at 37 °C with different concentrations of DHEA or EA by the MTT method and the CC₅₀ values for each compound were calculated (Table 1). Then, antiviral activity of DHEA and EA against the arenaviruses JUNV (strains XJCl3 and IV4454), TCRV and PICV was examined by a virus yield inhibition assay. Infected Vero cells were incubated, immediately after virus adsorption, in MM containing different DHEA or EA non-cytotoxic

Discussion

In the present study we demonstrated that the natural steroids DHEA, EA and three synthetic derivatives have a selective inhibitory action against arenaviruses. DHEA and EA did not exhibit virucidal activity even at concentrations five times higher than their EC₅₀ values. The inhibitory action of DHEA and EA along a single cycle of JUNV replication could not be attributed to a failure on virus adsorption or internalization. A strong inhibitory effect of these steroids on JUNV protein synthesis

Acknowledgements

We are grateful to Dr. A. Sanchez for his generous gift of monoclonal antibodies. This work was supported by grants from Agencia Nacional de Promoción Científica y Técnica (ANPCYT) PICT 12307/02 and from Universidad de Buenos Aires, UBA, X 046. E.G. Acosta is a fellow from the Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET, Argentina.

References (31)

V. Castilla et al.
Antiviral mode of action of a synthetic brassinosteroid against Junin virus replication
Antiviral Res.
(2005)
E. De Clercq
Antiviral drugs in current clinical use
J. Clin. Virol.
(2004)
G. Defaye et al.
Synthesis of 15β-hydroxylated derivatives of dehydroisoandrosterone and isoandrosterone
Tetrahedron Lett.
(1978)
D.A. Enria et al.
Antiviral treatment of Argentine hemorrhagic fever
Antiviral Res.
(1994)
R.M. Loria
Immune up-regulation and tumor apoptosis by androstene steroids
Steroids
(2002)
H. Nawata et al.
Mechanism of action of antiaging DHEA-S and the replacement of DHEA-S
Mech. Ageing Dev.
(2002)
C. Ruiz-Jarabo et al.
Lethal mutagenesis of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV)
Virology
(2003)
M. Tortorici et al.
Arenavirus nucleocapsid protein displays a transcriptional antitermination activity in vivo
Virus Res.
(2001)
S. Yoshida et al.
Anti-proliferative action of endogenous dehydroepiandrosterone metabolites on human cancer cell lines
Steroids
(2003)
W. Bradley et al.
Dehydroepiandrosterone inhibits replication of feline immunodeficiency virus in chronically infected cells
Vet. Immunol. Immunopathol.
(1995)

V. Castilla et al.

The entry of Junin virus into Vero cells

Arch. Virol.

(1994)

C. Chang et al.

Antiviral effect of dehydroepiandrosterone on Japanese encephalitis virus infection

J. Gen. Virol.

(2005)

S. Cooray

The pivotal role of phosphatidylinositol 3-kinase-Akt signal transduction in virus survival

J. Gen. Virol.

(2004)

E. Damonte et al.

Perspectives for the therapy against arenavirus infections

K. Diallo et al.

Inhibition of human immunodeficiency virus type-1 (HIV-1) replication by immunor (IM28), a new analog of dehydroepiandrosterone

Nucleosids Nucleotides Nucleic Acids

(2000)

Cited by (26)

Photocatalytic Three-Component Synthesis of 3-Heteroarylbicyclo[1.1.1]pentane-1-acetates
2023, Organic Letters
Herein, we report a visible-light-induced three-component reaction involving [1.1.1]propellane, diazoates, and various heterocycles for the synthesis of 3-heteroarylbicyclo[1.1.1]pentane-1-acetates. Throughout this reaction, the radicals generated from diazoate species react with [1.1.1]propellane in an addition reaction to form bicyclo[1.1.1]pentane (BCP) radicals that subsequently react with heterocycles, leading to the formation of 1,3-disubstituted BCP acetates. Notably, this methodology exhibits excellent functional group compatibility, high atom economy, and mild reaction conditions, thus facilitating suitable synthetic access to 1,3-disubstituted BCP acetates.
Screening of novel synthetic derivatives of dehydroepiandrosterone for antivirals against flaviviruses infections
2022, Virologica Sinica
Citation Excerpt :
However, persistent administration of DHEA leads to masculinization in women (Labrie et al., 2003). Previous studies have shown structural analogs of DHEA, which were used as potential therapeutic agents against human immunodeficiency virus 1, feline immunodeficiency virus, Junin virus, coxsackie B virus, herpes simplex type 2, vesicular stomatitis virus, JEV, and influenza virus (Acosta et al., 2008; Daigle and Carr, 1998; Pedersen et al., 2003). From primary screening, two analogs HAAS-AV3026 and HAAS-AV3027 displayed strong antiviral activity against JEV and ZIKV at non-cytotoxic concentrations and also exhibited higher CC50 values both in BHK-21 and Vero cell lines, and were chosen as the lead compound for downstream studies.
Flaviviruses are important arthropod-borne pathogens that represent an immense global health problem. Their unprecedented epidemic rate and unpredictable clinical features underscore an urgent need for antiviral interventions. Dehydroepiandrosterone (DHEA) is a natural occurring adrenal-derived steroid in the human body that has been associated in protection against various infections. In the present study, the plaque assay based primary screening was conducted on 32 synthetic derivatives of DHEA against Japanese encephalitis virus (JEV) to identify potent anti-flaviviral compounds. Based on primary screening, HAAS-AV3026 and HAAS-AV3027 were selected as hits from DHEA derivatives that exhibited strong antiviral activity against JEV (IC₅₀ = 2.13 and 1.98 μmol/L, respectively) and Zika virus (ZIKV) (IC₅₀ = 3.73 and 3.42 μmol/L, respectively). Mechanism study indicates that HAAS-AV3026 and HAAS-AV3027 do not exhibit inhibitory effect on flavivirus binding and entry process, while significantly inhibit flavivirus infection at the replication stage. Moreover, indirect immunofluorescence assay, Western blot analyses, and quantitative reverse transcription-PCR (qRT-PCR) revealed a potent antiviral activity of DHEA derivatives hits against JEV and ZIKV in terms of inhibition of viral infection, protein production, and viral RNA synthesis in Vero cells. Taken together, our results may provide a basis for the development of new antivirals against flaviviruses.
Synthesis and biological evaluation of D-ring fused 1,2,3-thiadiazole dehydroepiandrosterone derivatives as antitumor agents
2016, European Journal of Medicinal Chemistry
Citation Excerpt :
Dehydroepiandrosterone (DHEA) also known as androstenolone or prasterone, is the most abundant circulating steroid hormone in humans [11]. DHEA and its sulfate ester also have a variety of potential biological effects, such as anti-aging [12], anti-inflammatory [13], immunomodulatory [14], antiviral [15,16], antidepressant [17] and anticancer effects [18,19]. As heterocycles were fused with the D-ring of DHEA, the antitumor activity was greatly improved.
A series of D-ring fused 1,2,3-thiadiazole DHEA derivatives were synthesized and investigated for their activity against the growth of various tumor cell lines using the sulforhodamine B (SRB) assay. It is amazing that for these compounds, T47D cell line was much more sensitive than other tumor cell lines. The most potent saturated N-heterocyclic derivatives showed similar antitumor effect with the positive control compound ADM (adriamycin) on T47D cells, that was 44–60 folds more potent than the lead compound DHEA. Most compounds with potent antitumor activity displayed low toxicity on normal human fibroblasts (HAF). Especially compound 25 (CH33) showed an IC₅₀ of 0.058 μM on T47D cells and its selectivity index (SI) between HAF and T47D was 364, which was 214 folds better than ADM (SI = 1.7). The apoptosis, colony formation and transwell migration assays of 25 were performed on T47D cell line. The primary mechanism study showed that 25 caused a dose-dependent induction of apoptosis, and induced phosphorylation of EphA2 and EphB3 in T47D cells. The in vivo antitumor effect of 25 was also observed in T47D tumor-bearing mice without obvious toxicity.
Sterol analogues with diamide side chains interfere with the intracellular localization of viral glycoproteins
2012, Biochemical and Biophysical Research Communications
Citation Excerpt :
We have also found that compounds 1 and 2 (Fig. 1) exhibit anti herpes simplex virus 1 (HSV-1) activity in vitro and ameliorate the signs of murine herpetic stromal keratitis [5,6]. Similar studies performed in our laboratory have shown that dehydroepiandrosterone (3) and some synthetic derivatives display a broad spectrum of antiviral action against Junin virus, vesicular stomatitis virus (VSV) and adenovirus [7–9]. The actual knowledge about the antiviral action of these plant and animal steroids reveals that cellular targets are an important component of this activity, including modulation of cell signaling pathways [10].
The need to develop novel antiviral agents encouraged us to assess the antiviral activity of synthetic sterol analogues with a diamide side chains. Cytotoxicity and antiviral activity of a family of azasterol previously synthesized was evaluated against herpes simplex virus 1 (HSV-1) (KOS and B2006) and vesicular stomatitis virus (VSV). This family of compounds was extended by the synthesis of novel analogs using an Ugi multicomponent reaction and their ability to inhibit viral multiplication was also evaluated. The results show that some of the compounds tested exert an antiviral activity. Besides, the effect of the azasterols on the intracellular localization of viral glycoproteins was examined. Strikingly, alteration on the glycoprotein D (gD) of HSV-1 fluorescence pattern was observed with both the antiherpetic compounds and the inactive azasterols.
In vitro antiviral activity of dehydroepiandrosterone, 17 synthetic analogs and ERK modulators against herpes simplex virus type 1
2012, Antiviral Research
Citation Excerpt :
Several investigations are being made in order to identify DHEA analogs that keep its beneficial effects without being able to be transformed in sexual hormones. Many analogs have been tested as antiviral compounds in vitro and in vivo (Pedersen et al., 2003; Henderson et al., 1992; Diallo et al., 2000; Mavoungou et al., 2005; Acosta et al., 2008; Romanutti et al., 2009, 2010) and previous reports have shown in vitro antiherpetic activity for other steroidal compounds, specially against HSV-1 (Wachsman et al., 2000, 2004; Talarico et al., 2002). In order to regulate viral replication and host gene responses, many viruses are known to manipulate host-signaling machinery, including the mitogen-activated protein kinases (MAPKs) pathway.
In the present study the in vitro antiviral activity of dehydroepiandrosterone (DHEA) and 17 synthetic derivatives against herpes simplex type 1 (HSV-1) was determined. DHEA, epiandrosterone (EA), two synthetic DHEA analogs and three synthetic EA analogs showed a selective inhibitory effect on HSV in vitro multiplication. DHEA and E2, a synthetic derivative of EA, were not found to be virucidal to cell-free HSV-1 and did not impair virus adsorption or penetration. We determined that treatment with both compounds decreased viral protein synthesis. Moreover, inhibitory effect of DHEA and E2 on extracellular viral titer was stronger than the inhibition found on total viral infectivity, suggesting that the antiherpetic activity of these compounds may also be in part due to an inhibition in virus formation and release.
Since DHEA is a known Raf/MEK/ERK signaling pathway activator, we studied the role of this pathway on HSV-1 infection. ERK1/2 phosphorylation was stimulated in HSV-1 infected cultures. UO126, a Raf/MEK/ERK signaling pathway inhibitor, impaired viral multiplication, while anisomycin, an activator of this pathway, enhanced it.
Treatment with DHEA 6 h before infection enhanced HSV-1 multiplication. On the contrary, pre-treatment with E2, which does not modulate Raf/MEK/ERK signaling pathway, did not produce an increase of viral replication. Taking together these results, the antiviral activity of DHEA seems to occur via a mechanism independent of its ability to modulate ERK phosphorylation.
Screening and verification of antiviral compounds against HSV-1 using a method based on a plaque inhibition assay
2023, BMC Infectious Diseases

View all citing articles on Scopus

View full text

Dehydroepiandrosterone, epiandrosterone and synthetic derivatives inhibit Junin virus replication in vitro

Abstract

Introduction

Section snippets

Compounds

Antiviral activity of DHEA and EA against JUNV, PICV and TCRV

Discussion

Acknowledgements

Antiviral Res.

J. Clin. Virol.

Tetrahedron Lett.

Antiviral Res.

Steroids

Mech. Ageing Dev.

Virology

Virus Res.

Steroids

Dehydroepiandrosterone inhibits replication of feline immunodeficiency virus in chronically infected cells

Vet. Immunol. Immunopathol.

The entry of Junin virus into Vero cells

Arch. Virol.

Antiviral effect of dehydroepiandrosterone on Japanese encephalitis virus infection

J. Gen. Virol.

The pivotal role of phosphatidylinositol 3-kinase-Akt signal transduction in virus survival

J. Gen. Virol.

Perspectives for the therapy against arenavirus infections

Inhibition of human immunodeficiency virus type-1 (HIV-1) replication by immunor (IM28), a new analog of dehydroepiandrosterone

Nucleosids Nucleotides Nucleic Acids