Antiviral activity of SK&F 21681 against herpes simplex virus.

SK&F 21681 (3,10-dimethyl-10-H-s-triazolo[4',3':2,3]-as-triazino-[ 5,6-b]indole) is an inhibitor of the growth of herpes simplex viruses types 1 and 2 at a concentration of 60 micrograms/ml. It inhibits the synthesis of the viral DNA and the formation of virus particles, although the viral polypeptide synthesis is not significantly affected by this compound. Mutants of herpes simplex viruses types 1 and 2 which are able to grow in the presence of SK&F 21681 were isolated. They induced normal levels of thymidine kinase and DNA polymerase activities in the infected cells and did not show resistance to either 9-[2-hydroxyethoxymethyl] guanine or phosphonoacetic acid.

Antiviral compounds. 1. Structure-activity relationship of some antiviral enediones derived from aldehydo sugars.

A series of aldehydo sugars was subjected to condensation reactions with active methylene compounds. Acetylacetone was condensed with 2,4-O-benzylidene-3,5-O-dibenzoyl-D-ribose (1), 2,4:3,5-O-dibenzylidene-D-ribose (6), 2,3,4,5-tetraacetyl-D-ribose (7), and 2,3,4,5,6-pentaacetyl-D-glucose (9) to yield 3-ylidene-2,4-pentanedione derivatives 2, 11, 12, and 13, respectively. Sugar derivatives 1 and 6 were also condensed with benzoylacetone to give 14 and 18, with acetoacetanilide to give 16 and 19, with malononitrile to give 17 and 20, and with alpha-(gamma-butyrolactonylidene)triphenylphosphorane to give 21 and 22, respectively. Condensation of 1 with dibenzoylmethane gave 15. The double bond in compounds 2 and 11 was saturated by hydrogenation to give 23 and 24. All alpha, beta-unsaturated carbonyl compounds obtained exhibited antiviral activity and cytotoxicity. Compound 11 was found to have the most significant and selective antiviral activity against herpes simplex virus.

Characterization of an herpes simplex virus type 2 mutant, which is resistant to acycloguanosine and causes fusion of BSC1 cells.

A mutant of herpes simplex virus type 2, which induces low levels of thymidine-kinase activity in infected BSC1 cells and consequently able to grow in the presence of acycloguanosine, was isolated. This mutant has also been shown to cause fusion of BSC1 cells. In BSC1 cells, co-infected with the wild-type strain and the mutant, the yield of each of the two viruses was normal but the rounding and aggregation of cells observed, resembled that found in wild-type infected cultures. When the mixed infection was performed in the presence of acycloguanosine (100 micrometers), the growth of the two virus strains was inhibited, as well as the cytopathic effect in the cultures. It is suggested that under these conditions, the thymidine-kinase which was induced in the infected cells by the wild-type strain, phosphorylated acycloguanosine and the activated drug formed, inhibited the growth of the two viruses by interference in their DNA syntheses.

Inhibition of vaccinia virus maturation by zinc chloride.

Zinc chloride (0.1 mM) inhibited by 96.4% the growth of vaccinia virus in HeLa cells. Approximately 50% inhibition in formation of particles that sedimented in sucrose gradients similarly to vaccinia virions occurred in the presence of zinc ions. Whereas the synthesis of the viral deoxyribonucleic acid was not affected by zinc chloride, a decrease in the overall synthesis of viral polypeptides and inhibition of the cleavage of precursors to the core polypeptides were observed.

Interaction of Herpes simplex virus with human cell lines at various stages of lymphoid differentiation.

The growth of herpes simplex virus (HSV) type 1 in human lymphoid cell lines, arrested at various stages of differentiation, was studied. The synthesis of viral DNA and antigens and the production of infectious virus were followed in null, B and T leukemia-lymphoma cell lines. It was found that while "nondifferentiated" null cells and cells differentiating along the B pathway, even at very early stages of differentiation, supported HSV growth, cell lines of T origin at all stages of differentiation were generally nonpermissive. The failure of T cells to support the growth of HSV did not result from inefficient adsorption to the host cells. However, viral DNA synthesis was not detected, and a significant reduction in the ability of synthesize HSV antigens was observed. It is suggested that the block in the growth cycle of the virus occurs at a stage after adsorption of the virion to the cells, but prior to synthesis of its DNA and proteins.

Antiviral activity of tunicamycin on herpes simplex virus.

Tunicamycin (0.5 microgram/ml) significantly lowers (2 to 3 log10) the infectious yield of herpes simplex virus type 1 grown in chicken embryo fibroblasts and in BSC1 cells. Although virus particles are formed and the synthesis of the viral deoxyribonucleic acid is only partially affected by the antibiotic, the glycosylation of herpesvirus glycopeptides is amost completely inhibited. The morphology of virus particles made in the presence of tunicamycin is similar to that of intact virus particles, as demonstrated by electron microscopy. This suggests that the absence of the carbohydrate side-chain from the viral glycopeptides does not affect the overall integrity of the virion but decreases very significantly the infectivity of these particles.

A sensitive plaque inhibition technique for assay of antibodies to influenza virus: use to detect previous antigenic priming with influenza viruses.

Plaque reduction test performed in MDCK cells was found to be a more sensitive assay than hemagglutination inhibition, for revealing previous immunological experience with influenza A virus in the population. Based on the preimmunization antibody levels, determined by plaque reduction tests, it was possible to distinguish between previously primed and unprimed subjects among vaccinees.

Formation of vaccinia virus DNA-protein complex in the presence of isatin beta thiosemicarbazone (IBT).

The association of newly synthesized vaccinia virus DNA with proteins in infected HeLa cells was followed. A shift from a high density to a low density complex occurred between 3 and 6 h after infection. This process was not affected by isatin beta thiosemicarbazone (IBT), an inhibitor of pox virus growth. At 22 h after infection in the absence of IBT, mature virions of high density were observed; however, in the presence of the drug, high density DNA-protein complexes, which lack the two main virus core polypeptides, were formed.

Inhibition of vaccinia virus growth by the nucleoside analogue 1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide (virazole, ribavirin).

Virazole or Ribavirin (1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide) inhibits the growth of vaccinia virus at a concentration ode to a certain extent in the presence of Virazole, the DNA fails to acquire resistance to deoxyribonuclease and virus particles are not formed. Reversibility of the antiviral effect occurs when the drug is washed out from the infected cultures or when guanosine at an equimolar concentration is added.

The effect of tilorone hydrochloride on the growth of several animal viruses in tissue cultures.

Tilorone hydrochloride, at a concentration of 10 mug/ml inhibits the growth of herpes simplex virus type 1 in BS-C-1 cells. The growth of vaccinia virus in BS-C-1 cells is partially inhibited; however, six viruses containing RNA, including four members of the togavirus group grown in chick fibroblasts, are not affected by the drug. The inhibition of the growth of herpes virus by tilorone hydrochloride is greater when the multiplicity of infection is lower than 1 p.f.u/cell and when the drug is added early in the course of infection.

Inhibition of herpesvirus deoxyribonucleic acid and protein synthesis by tilorone hydrochloride.

Tilorone hydrochloride at a concentration of 10 mug/ml very efficiently inhibited herpes simplex virus growth in BSC1 cells when the virus is infected at a low multiplicity of infection. The adsorption of the virus was not affected by the drug, and the penetration of the deoxyribonucleic acid of the input virus into the cytoplasm and nuclei proceeded normally when tilorone hydrochloride was present. However, newly synthesized viral deoxyribonucleic acid was not detectable under these conditions, there was a remarkable decrease in the rate of viral polypeptide synthesis, and virus particles were not formed. The inhibition of herpesvirus growth by tilorone hydrochloride was absolutely dependent on the presence of the drug in the cultures. Pretreatment of the cells with the drug did not result in resistance to herpesvirus infection after the removal of the drug.

Inhibition of the growth of hemadsorption 2 virus by three acyl derivatives of amino acids.

Three acyl derivatives of amino acids, dicarbobenzoxy-l-lysine sodium, carbobenzoxy-l-aspartic acid-beta-benzyl ester potassium, and N-3-phenylpropionyl-S-benzyl-l-cysteine potassium inhibit the growth of parainfluenza 1 (hemadsorption 2) virus. The growth of simian virus 40, vaccinia, poliomyelitis type 1, Semliki Forest, Eastern equine encephalitis, and Western equine encephalitis viruses was not affected by these compounds. Four other acyl derivatives of amino acids did not inhibit the growth of any of the viruses tested.

Isolation and characterization of a gamma-thiochromanone-4-thiosemicarbazone-resistant mutant of vaccinia virus.

gamma-Thiochromanone-4-thiosemicarbazone (TCT) inhibits the growth of vaccinia virus in BSCl cells by interfering with viral maturation. A mutant of the virus (TCT(R)) which is resistant to this drug was isolated. This mutant also exhibits resistance to another thiosemicarbazone related compound, isatin beta-thiosemicarbazone (IBT). There is a good correlation between the cross-resistance of the two mutants IBT(R) and TCT(R) to TCT and IBT, respectively, and the similar antipoxvirus activity of these two thiosemicarbazone-related compounds.

Synthesis of vaccinia virus polypeptides in the presence of hydroxyurea.

Hydroxyurea (5 x 10(-3) M) inhibits the development of vaccinia virus in HeLa cells early after infection and prevents the synthesis of viral deoxyribonucleic acid. In the presence of hydroxyurea, late viral polypeptides are not formed, as observed by electrophoresis on polyacrylamide gels.