High-Throughput Screening and Identification of Human Adenovirus Type 5 Inhibitors.

Human adenovirus infections can develop into diffuse multi-organ diseases in young children and immunocompromised patients, and severe cases can lead to death. However, there are no approved antiviral drugs available to treat adenovirus diseases. In this study, a chemiluminescence-based, high-throughput screening (HTS) assay was developed and applied to screen human adenovirus 5(HAdV5)inhibitors from 1,813 approved drug library and 556 traditional Chinese medicine-sourced small-molecule compounds. We identified three compounds with in vitro anti-HAdV5 activities in the low-micromolar range (EC50 values 0.3-4.5 µM, selectivity index values 20-300) that also showed inhibitory effects on HAdV3. Cardamomin (CDM) had good anti-HAdV5 activity in vitro. Furthermore, three dilutions of CDM (150, 75, and 37.5 mg/kg/d) administered to BALB/c mouse models inhibited HAdV5-fluc infection at 1 day post-infection by 80% (p < 0.05), 76% (p < 0.05), and 58% (p < 0.05), respectively. HE-staining of pathological tissue sections of mice infected with a wildtype adenoviral strain showed that CDM had a protective effect on tissues, especially in the liver, and greatly inhibited virus-induced necrosis of liver tissue. Thus, CDM inhibits adenovirus replication in vivo and in vitro. This study established a high-throughput screening method for anti-HAdV5 drugs and demonstrated CDM to be a candidate for HAdV5 therapy, potentially providing a new treatment for patients infected with adenoviruses.

USC-087 protects Syrian hamsters against lethal challenge with human species C adenoviruses.

Human adenoviruses (AdV) cause generally mild infections of the respiratory and GI tracts as well as some other tissues. However, AdV can cause serious infection in severely immunosuppressed individuals, especially pediatric patients undergoing allogeneic hematopoietic stem cell transplantation, where mortality rates are up to 80% with disseminated disease. Despite the seriousness of AdV disease, there are no drugs approved specifically to treat AdV infections. We report here that USC-087, an N-alkyl tyrosinamide phosphonate ester prodrug of HPMPA, the adenine analog of cidofovir, is highly effective against multiple AdV types in cell culture. USC-087 is also effective against AdV-C6 in our immunosuppressed permissive Syrian hamster model. In this model, hamsters are immunosuppressed by treatment with high dose cyclophosphamide. Injection of AdV-C6 (or AdV-C5) intravenously leads to a disseminated infection that resembles the disease seen in humans, including death. We have tested the efficacy of orally-administered USC-087 against the median lethal dose of intravenously administered AdV-C6. USC-087 completely prevented or significantly decreased mortality when administered up to 4 days post challenge. USC-087 also prevented or significantly decreased liver damage caused by AdV-C6 infection, and suppressed virus replication even when administered 4 days post challenge. These results imply that USC-087 is a promising candidate for drug development against HAdV infections.

[Clinical efficacy of a herbal drug combination in acute viral rhinosinusitis]. / Klinische Wirksamkeit eines pflanzlichen Kombinationspräparates in der Behandlung der akuten viralen Rhinosinusitis.

BACKGROUND: Acute rhinosinusitis is a frequent inflammatory disease of the mucosa of the nose and paranasal sinuses, usually associated with substantial morbidity having considerable socioeconomic impact. A new herbal drug based on a dry extract of a combination of 5 medicinal drugs (Sinupret® extract Dragees) was tested in a confirmatory trial in patients with acute viral rhinosinusitis. METHODS: 386 patients with symptomatic acute viral rhinosinusitis have been treated with the herbal drug combination (daily dosage 3 × 160 mg) or placebo in a double-blind, randomised, placebo-controlled clinical trial for 15 days. Primary efficacy endpoint was the investigator assessed symptom score at the end of therapy (Major Symptom Score, MSSINV). RESULTS: Treatment with verum lead to a statistically significant, clinically relevant improvement of the symptom score (2.07 ± 0.18 [SEM] vs. 3.47 ± 0.28 score points, p = 0.0001; PP: N = 300) compared to placebo at visit 5. The Number Needed to Treat (NNT) was 7 (PP). Adverse events occurred in 9.8% of the patients treated with verum and 14.1% of the patients treated with placebo. No serious adverse event was observed. The investigators assessed the tolerability of the herbal drug combination predominantly as good and very good (96.4% verum, 95.3% placebo). CONCLUSION: The results prove the efficacy and tolerability of the herbal drug in the indication acute viral rhinosinusitis. Especially due to the favorable benefit-risk ratio the drug represents a suitable treatment alternative.

Disseminated adenovirus disease in heart transplant recipient presenting with conjunctivitis.

We report a 65-year-old heart transplant recipient who presented with conjunctivitis, likely acquired from a family member who worked at a daycare center during an outbreak of conjunctivitis. He developed a severe adenoviral pneumonitis, which was successfully treated with intravenous cidofovir combined with a reduction of immunosuppression.

Portuguese winemaking residues as a potential source of natural anti-adenoviral agents.

To date there are no licensed systemic or topical treatments in Europe or the USA for adenovirus infections. In the present paper, we evaluate the effect of a polyphenol-based grape extract (NE) obtained from Portuguese white-winemaking by-products, and Resveratrol in pure form, on adenovirus type 5 infection. For this purpose, recombinant adenovirus vectors (Ad-5) and a human-derived cell line (293) were used as models. The NE and Resveratrol at the used concentrations do not induce cell cytotoxicity or direct virucidal activity; however, they reduce 4.5 and 6.5 log (TCID(50)/ml) on total infectious Ad-5 production, respectively. The capacity of Ad-5 replication upon removal of NE and Resveratrol after 24 h post infection was also evaluated. In contrast to Resveratrol, the highest evaluated NE concentration inhibits irreversibly the Ad-5 replication. These results provide useful information for the use of NE and Resveratrol as potential sources of promising natural antiviral agents on Ad-5 infection.

Clinical and in vitro evaluation of cidofovir for treatment of adenovirus infection in pediatric hematopoietic stem cell transplant recipients.

Post-hematopoietic stem cell transplantation (HSCT) adenovirus infections were identified in 31 of 204 consecutive pediatric HSCT patients, 18 of whom had severe manifestations of infection. Cidofovir treatment led to clinical improvement in 8 of 10 patients with severe infection and to virologic clearance in 9 patients. In vitro susceptibility to cidofovir was demonstrated in 12 clinical adenovirus isolates. Cidofovir is a promising treatment option for this population.

[New medical treatment for viral conjunctivitis].

Around one million people in Japan are suffering from adenoviral conjunctivitis every year and it is recognized as one of the major pathogens of nosocomial infection. Several complications, such as corneal erosion and conjunctival pseudomembrane, are observed in some of the cases and corneal sube- pithelial opacity may bring visual impairment. Moreover, no specific anti-adenoviral agent has been discovered and an effective treatment has not been established for adenoviral infection. We have researched new medical treatment for viral conjunctivitis based on recent findings in adenoviral conjunctivitis. Firstly, anti-adenoviral activity was evaluated in vitro in agents which could possibly act as anti-adenoviral drugs. Twelve candidates, such as zalcitabin, interferon beta, etc., were selected among antiviral drugs, adenoviral receptor inhibitors, natural products and anti-inflammatory drugs. Remarkable anti-adenoviral effect was observed in zalcitabin, sanilbudine, interferon beta and anti-osteopontin peptide. Two anti-human immunodeficiency virus (HIV) drugs with anti-adenoviral activity, zalcitabin and sanilbudine, are nucleoside reverse transcriptase inhibitors, but, in contrast, non-nucleoside reverse transcriptase inhibitors and protease inhibitors were ineffective against adenovirus. Interferon beta and anti-osteopontin peptide displayed anti-adenoviral effects by absorption inhibition. Secondly, side effects caused by possible anti-adenoviral eye drops, including cidofovir whose development as eye drops against eyeball and ocular adnexa had been suspended, were analyzed in a white rabbit model. In animals given cidofovir locally, significant narrowing of lacrimal canaliculus, redness of eyelid and conjunctival injection was observed, but obstruction of the lacrimal duct was not found. Although zalcitabin and sanilbudine eye drops induced eyelid redness, no change was observed in the lacrimal route and conjunctiva. In animals treated by cidofovir, inflammation histologically suggesting mainly allergic change was observed. These results indicate that these four drugs are possible candidate for safe eye drops against adenoviral conjunctivitis. These four agents are divided into two categories, inhibitors of adenoviral replication, zalcitabin and sanilbudine; and suppressors of adenoviral infection, interferon beta and anti-osteopontin peptide. It is expected that eye drops for specific treatment of adenoviral conjunctivitis are going to be available in the near future following investigation of therapeutic effect in adenoviral infected animals and clinical trials in humans.

239 cases of high fever in viral upper respiratory infection (URI) treated with xiang shi qing jie (XSQJ) bag tea.

XSQJ bag tea was used to treat 239 cases of high fever caused by viral upper respiratory infection, widely spreading in summer and autumn. For comparison, a control group treated by penicillin combined with Ganmao Qingre Chongji or Banlangen Chongji was set up. The results showed the effect of XSQJ bag tea on the time of abating fever, resolving symptoms and physical signs were significantly better than that of the control group.

[Effect of antioxidants on the HeLa cell infection process in culture]. / Vliianie antioksidantov na protsess infitsirovaniia kletok HeLa v kul'ture.

A certain role in the outcome of virus infection of cells was assumed to belong to cell membrane lipids peroxidation. The influence of known inhibitors of lipid peroxidation, beta-naphthol and ionol, on HeLa cells infection with human adenovirus type 2 was studied. These antioxidants in certain concentrations were found to be capable of effectively inhibiting virus infection of HeLa cell cultures. At the same time, beyond these concentration levels these antioxidants did not inhibit but stimulated virus infection of HeLa cell cultures. It is concluded that inhibition of virus infection of cells by antioxidants in moderate concentrations is due to their blocking of membrane lipids peroxidation. The opposite effect of antioxidants in much higher or lower concentrations may be due to their membranotropic effect which enhances the development of virus infection. The results of the study attest to the possible use of antioxidants in appropriate concentrations for control of virus infection.