Screening for anti-HIV drugs that can combine virucidal and virustatic activities synergistically.

Abstract: Hepatitis C virus (HCV) infection affects an estimated 185 million people worldwide, with chronic infection often leading to liver cirrhosis and hepatocellular carcinoma. Although HCV is curable, there is an unmet need for the development of effective and affordable treatment options. Through a cell-based high-throughput screen, we identified chlorcyclizine HCl (CCZ), an over-the-counter drug for allergy symptoms, as a potent inhibitor of HCV infection. CCZ inhibited HCV infection in human hepatoma cells and primary human hepatocytes. The mode of action of CCZ is mediated by inhibiting an early stage of HCV infection, probably targeting viral entry into host cells. The in vitro antiviral effect of CCZ was synergistic with other anti-HCV drugs, including ribavirin, interferon-α, telaprevir, boceprevir, sofosbuvir, daclatasvir, and cyclosporin A, without significant cytotoxicity, suggesting its potential in combination therapy of hepatitis C. In the mouse pharmacokinetic model, CCZ showed preferential liver distribution. In chimeric mice engrafted with primary human hepatocytes, CCZ significantly inhibited infection of HCV genotypes 1b and 2a, without evidence of emergence of drug resistance, during 4 and 6 weeks of treatment, respectively. With its established clinical safety profile as an allergy medication, affordability, and a simple chemical structure for optimization, CCZ represents a promising candidate for drug repurposing and further development as an effective and accessible agent for treatment of HCV infection.

Abstract: Diclofenac sodium (Dc), an anti-inflammatory agent, has remarkable inhibitory action both against drug-sensitive and drug-resistant clinical isolates of various Gram-positive and Gram-negative bacteria. The aim of this study was to determine the ability of Dc to protect mice from a virulent Salmonella infection. Dc injected at 1.5 microg/g and 3.0 microg/g mouse body weight significantly protected animals from the lethality of Salmonella infection. As was the case for the in vitro interaction, Dc in combination with streptomycin was even more effective. The non-antibiotic drug Dc has potential for the management of problematic antibiotic-resistant bacterial infections.

Abstract: Diclofenac sodium (Dc) was found to possess antibacterial activity against both drug-sensitive and drug-resistant clinical isolates of Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, and Mycobacterium spp., in addition to its potent anti-inflammatory activity. The time-kill curve study indicates that this non-steroidal drug exhibits bactericidal activity against Listeria, E. coli, and M. tuberculosis. The antibacterial activity of Dc comes, in part, from its ability to inhibit the DNA synthesis of E. coli and L. monocytogenes. Dc could protect murine listeriosis, salmonellosis, and tuberculosis at doses ranged within its maximum recommended human or non-toxic ex-vivo dose. Dc possesses anti-plasmid activity and acts as a ‘helper compound’ in synergistic combination with streptomycin against E. coli and Mycobacterium or gentamicin against Listeria. This review focuses on the possible use of Dc, a non-antibiotic helper compound, in infections and inflammatory conditions, rationalized on the basis of the activities of the compounds.

Abstract: A body of evidence supports an association between Chlamydia pneumoniae and atherosclerosis. Recent prospective, seroepidemiologic studies have refined estimations of relative risk. Advances in diagnostic testing with the polymerase chain reaction have created a potential opportunity to screen for infected individuals. New insights into the pathogenesis of infection with C. pneumoniae have been reported, many of which are relevant to the development of atherosclerotic plaque. Clinical trials have now been initiated and should provide guidance as to the utility of antibiotics in the treatment or prevention of coronary artery disease.

Abstract: The NS2B-NS3 protease has been identified as an attractive target for drug development against Zika virus (ZIKV) and combined drug repurposing and structure-based virtual screening has improved the development of antiviral drugs. In this study, we performed a structure-based virtual screening of 1861 Food and Administration (FDA) approved drugs available in DrugBank by the selection and docking validation of crystal structure of ZIKV NS2B-NS3 protease (PDB ID 5H4I ) using Glide and DOCK 6 software. The antihistaminic chlorcyclizine (Grid score -24.8 kcal/mol) exhibited the most promising interaction with NS2B-NS3 protease in comparison to crystallography ligand (Grid score -15.6 kcal/mol) by interaction to Tyr161 by hydrophobic interactions in the binding site of NS2B-NS3 which is recognized as an important amino acid in substrate molecular recognition. Cytotoxicity and global antiviral activity assay in Vero cells by MTT method showed that chlorcyclizine reduced the ZIKV induced cytopathic effect (EC50 of 69.0 ± 7.3 μM and SI = 1.9), and explicit molecular dynamics simulations implemented on a NAMD program indicated great stability of chlorcyclizine in protease binding site, suggesting the repurposing of chlorcyclizine as a promising finding in anti-ZIKV drug development.

Abstract: We previously described a set of four strains of Salmonella typhimurium designed for detecting the various types of mutagens, and showed their utility in detecting a wide variety of carcinogens as mutagens. The lipopolysaccharide that normally coats these bacteria is a barrier to penetration of mutagens to the cell membrane. The set of tester strains has been improved by adding a mutation (rfa: deep rough) that results in a deficient lipopolysaccharide. The techniques for using these strains for detecting mutagens are presented and the tests are shown to be extremely sensitive and convenient. The specificity of frameshift mutagenesis is clarified. As adjuncts to the test with the four strains, we describe a test that compares mutagenic killing in deep rough strains with and without DNA excision repair, and a test using forward mutagenesis in a deep rough strain lacking excision repair.

Abstract: This book presents the latest information from leading authorities involved in ongoing AIDS research and developments. Featured in this new edition are chapters on the laboratory diagnosis of the HIV infection; TB and other bacterial infections; cytomegaloviruses and other infections; Kaposi's sarcoma in AIDS; neurological manifestation in the HIV infected patient; counselling of the HIV-infected individual, their families and partners; and risk exposure to the health care worker.

Abstract: Mice infected with the LP-BM5 murine leukemia virus (MuLV) develop an immunodeficiency syndrome (murine AIDS) and an encephalopathy characterized by impaired spatial learning and memory. Because platelet-activating factor (PAF) has been implicated in the pathogenesis of HIV-associated dementia complex, brain PAF levels were measured in LP-BM5 MuLV-infected mice. PAF levels in cerebral cortex and hippocampus were significantly increased at 6 and 12 weeks after LP-BM5 MuLV inoculation, whereas significant increases in striatal and cerebellar PAF levels were observed only at 12 weeks after inoculation. Administration of the NMDA antagonist MK-801 significantly reduced the increased PAF levels in the cerebral cortex and hippocampus of LP-BM5 MuLV-infected mice. These results indicate that the LP-BM5 MuLV-induced increases in brain PAF levels are the results of NMDA receptor activation and are consistent with the hypothesis that elevated CNS PAF levels contribute to the behavioral deficits observed in LP-BM5 MuLV-infected mice.

Abstract: Publisher Summary This chapter describes the technique of correlative assays The correlative assays are used quite frequently to monitor the titers of fermentation liquors during culture development and during the isolation process The first step in the development of a correlative assay involves the testing of the compounds of interest against a broad spectrum of different microorganisms If a strong enough activity is found, a correlative microbiological assay can be developed to replace the original assay The chapter presents a list of microorganisms used in the correlative testing program and summarizes the specific cultivation conditions for each of the microorganisms used in the program A specific example is used to demonstrate the method of establishing the correlation between the primary biological activity and the correlative antimicrobial activity The most sensitive among the microorganisms used in this program appears to be Lactobacillus casei It is inhibited by more than 50% of the samples tested The next most sensitive microorganism was Bacillus subtilis, particularly when cultivated in a completely synthetic growth medium The chapter also discusses several advantages of microbiological assays