Detection of Inhibition of Bovine Viral Diarrhea Virus by Aromatic Cationic Molecules

Abstract

Bovine viral diarrhea virus (BVDV) is an economically significant pathogen of cattle and a problematic contaminant in the laboratory. BVDV is often used as an in vitro model for hepatitis C virus during drug discovery efforts. Aromatic dicationic molecules have exhibited inhibitory activity against several RNA viruses. Thus, the purpose of this research was to develop and apply a method for screening the aromatic cationic compounds for in vitro cytotoxicity and activity against a noncytopathic strain of BVDV. The screening method evaluated the concentration of BVDV in medium and cell lysates after 72 h of cell culture in the presence of either a 25 or 5 M concentration of the test compound. Five of 93 screened compounds were selected for further determination of inhibitory (90 and 50%) and cytotoxic (50 and 10%) concentration endpoints. The screening method identified compounds that exhibited inhibition of BVDV at nanomolar concentrations while exhibiting no cytotoxicity at 25 M concentrations. The leading compounds require further investigation to determine their mechanism of action, in vivo activity, and specific activity against hepatitis C virus.Bovine viral diarrhea virus (BVDV), which is the prototype of the Pestivirus genus of the Flaviviridae family, causes early embryonic death, abortion, teratogenesis, respiratory problems, chronic wasting syndrome, and immune system dysfunction in cattle throughout the world. Acute infections of immunocompetent cattle with different strains of BVDV have caused mortality rates of 17 to 32% (8,15,31). Vaccines have provided inconsistent protection against infection with BVDV (36).In addition to being a pathogen of cattle, BVDV can be a problematic contaminant in the laboratory. Two biotypes of BVDV (noncytopathic and cytopathic), which are based on their effects in cell cultures, are recognized (3). Noncytopathic biotypes of BVDV have been identified in commercially available lots of fetal bovine serum despite testing by the manufacturer (6, 45). Consequently, BVDV has been identified in commercially available bovine, canine, feline, and primate cell lines (16,19), human viral vaccines for measles-mumps-rubella (18, 21), and interferons for human use (20). Viral contamination of biologicals for human use may be the reason BVDV-specific antibodies have been found in some samples of human serum (17). No antiviral pharmaceuticals are currently available for controlling BVDV in the laboratory or on the farm.Hepatitis C virus (HCV), a member of the Hepacivirus genus of the family Flaviviridae, is a major cause of human liver disease throughout the world. The World Health Organization estimates that 170 million people are chronically infected with HCV (2). The organization of the HCV genome encoding the proteins for viral replication is very similar to that of BVDV (with the exception of the 5Ј-terminal protease) (4). The inability to propagate HCV efficiently by cell culture has caused researchers to adopt BVDV as a viral model for HCV (2). Because of the relatedness of BVDV...

Effect of Hydrogen Bonding on the Pyramidalization of the Amino Group: Structure of 3,4-diaminobenzamidinium Chloride

“…1 The most studied DNA minor groove binders are aromatic amidines with cationic amidine moiety at terminal part of molecules. A large number of compounds from this class which comprise an amidino substituted benzimidazole ring show a potent antimicrobial, [2][3][4][5] antiviral 6,7 and anticancer activity. [8][9][10] The most convenient method of benzimidazole synthesis includes the condensation of 3,4-diaminobenzamidine with aldehyde in the presence of a suitable oxidative reagent such as: 1,4-benzoquinone, 10,11 sodium bisulfate, 12,13 ceric ammonium nitrate 14 or nitrobenzene.…”

The Synthesis of Dicationic Extended Bis-Benzimidazoles

“…The compounds were inactive against E. coli, but showed good anti-Bacillus activity as indicated in Table 1. The compounds were also tested for mammalian cell in vitro cytotoxicity using CV1 Vero cells with the XTT cytotoxicity test (Table 1) [7]. A desirable therapeutic index (2 log differential) was noted for 3c.…”

“…[13]. In vitro cell toxicity tests were performed as described in the literature [7] using CV1 cells cultured in Glutamax RPMI medium (Invitrogen) plus 5% fetal bovine serum (Atlanta Biologicals), 100 U of penicillin G/mL, 100 µg streptomycin/mL and 0.25 µg of amphotericin B/mL. XTT was from Sigma.…”

Molecular Modeling of Small Molecules as BVDV RNA-Dependent RNA Polymerase Allosteric Inhibitors

“…Discovery of small molecule inhibitors of BVDV RdRp as a potential therapeutic target has been reported in the literature with various scaffolds, such as imidazopyridines, [7][8][9][10][11] benzimidazole derivatives, 12,13 arylazoenamines, 14,15 indole derivatives, 16 γ-carboline derivatives, 17 thiosemicabarzone, 18 diphenylmethane, 19,20 and aromatic cationic molecules. 21,22 The majority of anti-BVDV inhibitors could also be taken as an accurate measurement of antiviral activity against HCV or other Pestiviruses (e.g., CSFV, BDV) and Flaviviruses (e.g., YFV, WNV, DENV) in the same family for the purpose of exploiting approved treatments. However, the direct relationship between anti-BVDV activity and HCV blocking activity is not known, if BVDV was used as a surrogate system for HCV.…”