Construction of bridged polycyclic systems via radical cyclizations. Uncovering of a novel carbocyclization-ring expansion sequence.

[reaction: see text]. While exploring the application of radical cyclizations to the construction of bridged polycyclic systems, we have discovered a novel tandem process consisting of an endo-cyclization of a vinyl radical onto a terminal alkene followed by a one-carbon ring expansion promoted by the resulting secondary radical.

Straightforward Construction of Fused 6,7,5-Tricarbocyclic Systems by Tandem [5 + 2]/[4 + 2] Cycloadditions.

One-pot coupling of an intramolecular thermal [5C + 2C] pyrone-alkene cycloaddition to a [4C + 2C] Diels-Alder reaction provides immediate access to 6,7,5-tricarbocyclic systems bearing a 1,4-oxa-bridge in the seven-membered carbocycle. The transformation entails the net formation of four carbon-carbon bonds and creates three new cycles and a minimum of five new stereocenters. Preliminary attempts to open the oxa-bridge by reaction of the adducts with samarium diiodide and trimethylsilyl triflate led to relatively unexpected deoxygenated and aromatized products.

Use of persistent infections with vaccinia virus recombinants to introduce alterations in foreign proteins: an application to HIV-1 env protein.

With the aim of generating a virus-cell system to introduce alterations in proteins of interest--which may be of use in studies of their biological functions--we established a persistent infection on a B-lymphoma cell line (A20.2J) with vaccinia virus (VV) recombinants. As a model, we used a vaccinia virus recombinant expressing the human immunodeficiency virus HIV-1 env gene. In this unique virus-cell system, we found that it is possible to introduce several structural and functional alterations in the env protein with passage numbers. From passage 10-20, two new env products emerged: an uncleaved gp160 and a glycoprotein fragment of 110 kDa. The uncleaved gp160 exhibit interesting properties as an immunogen. This protein forms stable oligomers, is not released from the cells, cannot fuse CD4+ presenting HeLa cells and activates a stronger cellular immune response than the parental cleaved env. In contrast, the 110 kDa product is a poor immunogen, since it lacks the gp41 domain, cannot form oligomers, accumulates intracellularly and cannot fuse CD4+ cells. In the persistently infected cells we have also found alterations in another heterologous protein-beta-galactosidase-a gene inserted in the same locus of VV as the env gene. This alteration resulted in a truncation of the (beta-galactosidase protein from 125 kDa to about 70 kDa. A similar size truncation of env and of beta-galactosidase was observed in many of the isolated VV recombinants.