Inhibition of Human Rhinovirus-Induced Cytokine Production by AG7088, a Human Rhinovirus 3C Protease Inhibitor

Zalman, Leora S.; Brothers, Mary A.; Dragovich, Peter S.; Zhou, Ru; Prins, Thomas J.; Worland, Stephen T.; Patick, Amy K.

doi:10.1128/aac.44.5.1236-1241.2000

Cited by 54 publications

(44 citation statements)

References 39 publications

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“…These data extend the work of Zalman and colleagues (32), who examined the requirement of 3C protease for airway epithelial cell responses. In BEAS-2B cells, treatment with AG7088, an inhibitor of 3C protease, inhibited the production of IL-6 and IL-8.…”

Section: Discussionsupporting

confidence: 72%

Rhinovirus 16 3C Protease Induces Interleukin-8 and Granulocyte-Macrophage Colony-Stimulating Factor Expression in Human Bronchial Epithelial Cells

et al. 2004

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Rhinovirus (RV), a member of the Picornaviridae family, accounts for many virus-induced asthma exacerbations. RV induces airway cell chemokine expression both in vivo and in vitro. Because of the known interactions of proteases with cellular functions, we hypothesized that RV 3C protease is sufficient for cytokine up-regulation. A cDNA encoding RV16 3C protease was constructed by PCR amplification and transfected into 16HBE14oϪ human bronchial epithelial cells. 3C protease induced expression of both IL-8 and GM-CSF, as well as transcription from both the IL-8 and GM-CSF promoters. 3C expression also induced activator protein 1 and NF-B transcriptional activation. Finally, mutation of IL-8 promoter AP-1 and NF-B promoter sequences significantly reduced 3C-induced responses. Together, these data suggest expression of RV16 3C protease is RVs are the most common upper respiratory pathogens, inducing the majority of common colds worldwide. Viral infections trigger asthma in 80 to 85% of asthma exacerbations in children and 44% in adults (1, 2) RV, a member of the Picornaviridae family of small, positive-stranded RNA viruses, accounts for much of virus-induced asthma exacerbations (1, 2). Cytokines are elaborated in vivo in the nasal mucosa, and most likely also from bronchial epithelium, after RV infection. Neutrophils, eosinophils, and lymphocytes increase in the nasal secretions of RV-infected patients compared with uninfected controls (3, 4).The cytokines IL-8 and GM-CSF have been implicated in the pathogenesis of asthma. IL-8 is a potent activator and chemoattractant of neutrophils (5) and eosinophils (6), and GM-CSF promotes the differentiation and survival of recruited eosinophils (7). Increased levels of IL-8, GM-CSF (8), and ICAM-1 (9), the receptor of 90% of RV serotypes, as well as IL-1 and IL-6, have been found in the nasal secretions of individuals infected with RVs (10 -12). In bronchial mucosal biopsies obtained after experimental infection with RV, Bardin and colleagues (13) noted a significant increase in submucosal lymphocytes and epithelial eosinophils, which, in asthmatics, persisted into convalescence. Although increases in cytokine secretion in the nasal mucosa does not prove increased expression in the epithelium in the lower airways, taken together the above data strongly suggest that cytokine elaboration is increased in the RV-infected lower airway epithelium.RV infection also induces cytokine production and adhesion molecule expression in cell culture. Divergent findings regard-

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Section: Discussionsupporting

confidence: 72%

Rhinovirus 16 3C Protease Induces Interleukin-8 and Granulocyte-Macrophage Colony-Stimulating Factor Expression in Human Bronchial Epithelial Cells

et al. 2004

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“…Initial sequence comparisons of the 3C protease-coding regions from six HRV serotypes (15,28,38,57,58,63), together with the experimentally derived three-dimensional structure of 3C protease (41), indicate significant homology in the substrate-inhibitor binding site, including strict conservation of the three amino acid residues that comprise the catalytic triad (His 40, Glu 71, and Cys 147). Consistent with this finding, rupintrivir (formerly AG7088), a novel, irreversible inhibitor of 3C protease (10-13, 61) has demonstrated broad-spectrum, potent in vitro antiviral activity against all picornaviruses tested, including 48 HRV serotypes, 4 HEV strains, and 46 untyped field isolates of HRV (34,46,64). Furthermore, recent data demonstrating the ability of rupintrivir to moderate illness severity and reduce viral load in human subjects following experimental HRV infection provide proof of concept for the mechanism of 3C protease inhibition (25).…”

mentioning

confidence: 49%

Conservation of Amino Acids in Human Rhinovirus 3C Protease Correlates with Broad-Spectrum Antiviral Activity of Rupintrivir, a Novel Human Rhinovirus 3C Protease Inhibitor

Binford

Maldonado

Weady

et al. 2005

Antimicrob Agents Chemother

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142

116

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The picornavirus 3C protease is required for the majority of proteolytic cleavages that occur during the viral life cycle. Comparisons of published amino acid sequences from 6 human rhinoviruses (HRV) and 20 human enteroviruses (HEV) show considerable variability in the 3C protease-coding region but strict conservation of the catalytic triad residues. Rupintrivir (formerly AG7088) is an irreversible inhibitor of HRV 3C protease with potent in vitro activity against all HRV serotypes (48 of 48), HEV strains (4 of 4), and untyped HRV field isolates (46 of 46) tested. To better understand the relationship between in vitro antiviral activity and 3C protease-rupintrivir binding interactions, we performed nucleotide sequence analyses on an additional 21 HRV serotypes and 11 HRV clinical isolates. Antiviral activity was also determined for 23 HRV clinical isolates and four additional HEV strains. Sequence comparison of 3C proteases (n ‫؍‬ 58) show that 13 and 11 of the 14 amino acids that are involved in side chain interactions with rupintrivir are strictly conserved among HRV and HEV, respectively. These sequence analyses are consistent with the comparable in vitro antiviral potencies of rupintrivir against all HRV serotypes, HRV isolates, and HEV strains tested (50% effective concentration range, 3 to 183 nM; n ‫؍‬ 125). In summary, the conservation of critical amino acid residues in 3C protease and the observation of potent, broad-spectrum antipicornavirus activity of rupintrivir highlight the advantages of 3C protease as an antiviral target.The picornavirus family consists of over 200 medically important viruses, including human rhinoviruses (HRV) and human enteroviruses (HEV). HRV, comprising over 100 different serotypes, are a major cause of mild upper respiratory infections (reviewed in references 4, 8, and 40). Although HRV infections are usually mild and self-limiting, they can also be associated with exacerbation of disease in individuals with underlying respiratory disorders (4, 48). The HEV include over 70 viruses that are associated with diverse clinical syndromes ranging from mild, self-limiting infections to fulminant and potentially fatal disease (3,52,53). Earlier clinical studies with agents that inhibit virus attachment and/or uncoating (e.g., tremacamra, a soluble intracellular adhesion molecule-1, pirodavir, and pleconaril) and nonspecific antiviral agents such as ␣-2␤ interferon have demonstrated that prevention and early treatment of HRV colds could provide clinical benefit (3, 18-23, 53). Recently, a retrospective analysis of two multicenter clinical trials demonstrated that pleconaril, a capsid-function inhibitor (17), significantly reduced the duration and severity of picornavirus-induced colds (24). To date, however, no antiviral agents have been approved for the prevention or treatment of HRV infection.We have focused our antiviral strategy on the inhibition of 3C protease, a viral enzyme that is absolutely required for the proteolytic cleavage of viral precursor polyproteins to functional p...

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“…Recognition of double-stranded RNA by protein kinase R has been proposed as another possible mechanism leading to cytokine production, but there is no clear evidence at this time for involvement protein kinase R in cytokine induction after RV infection (15). Also, RV 3C protease was reported to be important in NF-B activation and cytokine production from RV-infected epithelial cells (13,66). In monocytes/M, transient degradation of IB was shown to take place and suggested to be important for MCP-1 production (21).…”

Section: Discussionmentioning

confidence: 99%

Rhinovirus Replication in Human Macrophages Induces NF-κB-Dependent Tumor Necrosis Factor Alpha Production

Laza-Stanca

Stanciu

Message

et al. 2006

J Virol

100

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Rhinoviruses (RV) are the major cause of acute exacerbations of asthma and chronic obstructive pulmonary disease (COPD). Rhinoviruses have been shown to activate macrophages, but rhinovirus replication in macrophages has not been reported. Tumor necrosis factor alpha (TNF-␣) is implicated in the pathogenesis of acute exacerbations, but its cellular source and mechanisms of induction by virus infection are unclear. We hypothesized that rhinovirus replication in human macrophages causes activation and nuclear translocation of NF-B, leading to TNF-␣ production. Using macrophages derived from the human monocytic cell line THP-1 and from primary human monocytes, we demonstrated that rhinovirus replication was productive in THP-1 macrophages, leading to release of infectious virus into supernatants, but was limited in monocytederived macrophages, likely due to type I interferon production, which was robust in monocyte-derived but deficient in THP-1-derived macrophages. Similar to bronchial epithelial cells, only small numbers of cells supported complete virus replication. We demonstrated RV-induced activation of NF-B and colocalization of p65/NF-B nuclear translocation with virus replication in both macrophage types. The infection induced TNF-␣ release in a time-and dose-dependent, RV serotype-and receptor-independent manner and was largely (THP-1 derived) or completely (monocyte derived) dependent upon virus replication. Finally, we established the requirement for NF-B but not p38 mitogen-activated protein kinase in induction of TNF-␣. These data suggest RV infection of macrophages may be an important source of proinflammatory cytokines implicated in the pathogenesis of exacerbations of asthma and COPD. They also confirm inhibition of NF-B as a promising target for development of new therapeutic intervention strategies.

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Inhibition of Human Rhinovirus-Induced Cytokine Production by AG7088, a Human Rhinovirus 3C Protease Inhibitor

Cited by 54 publications

References 39 publications

Rhinovirus 16 3C Protease Induces Interleukin-8 and Granulocyte-Macrophage Colony-Stimulating Factor Expression in Human Bronchial Epithelial Cells

Rhinovirus 16 3C Protease Induces Interleukin-8 and Granulocyte-Macrophage Colony-Stimulating Factor Expression in Human Bronchial Epithelial Cells

Conservation of Amino Acids in Human Rhinovirus 3C Protease Correlates with Broad-Spectrum Antiviral Activity of Rupintrivir, a Novel Human Rhinovirus 3C Protease Inhibitor

Rhinovirus Replication in Human Macrophages Induces NF-κB-Dependent Tumor Necrosis Factor Alpha Production

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