We have looked for conserved DNA sequences between four herpes simplex virus type 1 (HSV-1) glycoprotein genes encoding gB, gC, gD, and gE and pseudorabies virus (PRV) DNA, HSV-1 DNA fragments representing these four glycoprotein-coding sequences were hybridized to restriction enzyme fragments of PRV DNA by the Southern blot procedure. Specific hybridization was observed only when HSV-1 gB DNA was used as probe. This region of hybridization was localized to a 5.2-kilobase (kb) region mapping at approximately 0.15 map units on the PRV genome. Northern blot (RNA blot) analysis, with a 1.2-kb probe derived from this segment, revealed a predominant hybridizing RNA species of approximately 3 kb in PRV-infected PK15 cells. DNA sequence analysis of the region corresponding to this RNA revealed a single large open reading frame with significant nucleotide homology with the gB gene of HSV-1 KOS 321. In addition, the beginning of the sequenced PRV region also contained the end of an open reading frame with amino acid homology to HSV-1 ICP 18.5, a protein that may be involved in viral glycoprotein transport. This sequence partially overlaps the PRV gB homolog coding sequence. We have shown that the PRV gene with homology to HSV-1 gB encoded the gII glycoprotein gene by expressing a 765-base-pair segment of the PRV open reading frame in Escherichia coli as a protein fused to beta-galactosidase. Antiserum, raised in rabbits, against this fusion protein immunoprecipitated a specific family of PRV glycoproteins of apparent molecular mass 110, 68, and 55 kilodaltons that have been identified as the gII family of glycoproteins. Analysis of the predicted amino acid sequence indicated that the PRV gII protein shares 50% amino acid homology with the aligned HSV-1 gB protein. All 10 cysteine residues located outside of the signal sequence, as well as 4 of 6 potential N-linked glycosylation sites, were conserved between the two proteins. The primary protein sequence for HSV-1 gB regions known to be involved in the rate of virus entry into the cells and cell-cell fusion, as well as regions known to be associated with monoclonal antibody resistance, were highly homologous with the PRV protein sequence. Furthermore, monospecific antibody made against PRV gII immunoprecipitated HSV-1 gB from infected cells. Taken together, these findings suggest significant conservation of structure and function between the two proteins and may indicate a common evolutionary history.

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J. Med. Chem. ester was the sole product. Pure 23 was obtained by hydrolysis with 0.01 N HCl in THF at 25 "C for 15 min.M solution in Krebs bicarbonate buffer saturated with C02 (pH 7.4) at 37 "C and measuring the capacity of this solution at 4-h intervals to cause relaxation of renal mesenteric artery previously contracted by PGF2,. After 24 h, relaxation had decreased by 50%, while PG12 under identical conditions required only 10 min.29 In these experiments, 1 and PG12 showed ECm values of 2.3 f 0.5 X lo4 and 3.3 f 0.5 X lo4 M, respectively.30 Relaxation of bovine coronary artery is uniquely characteristic for PG12 among all the pr~staglandins.~~ In this assay, 1 and PG12 showed ECm = 8.5 f 1.6 X lo4 and 2.8 f 0.6 X respectively.B Comparison of the potency of 1 and PG12 in causing complete inhibition of ADP and arachidonic acid induced aggregation of human platelets showed 1 (EDloo = lo-* M) to be 70% as active as PG12.1932 Intravenous administration of 1 and PGIz in doses of 1 to 2 X mol/kg as a bolus to an anesthetized dog showed the difluoro derivative to be equal in potency to the natural product in lowering blood pressure and decreasing peripheral and increasing renal blood flow.% Only a t the highest levels of 1 was a two-to threefold prolongation of action observed when 1 was compared with equipotent levels of PGIF Since 1 was shown to be completely resistant t o 15-hydroxyprostaglandin dehydrogenase,% rapid excretion either unchanged or after 8and/or P-450 catalyzed oxidation is probable. 10,10-Difluoro-13-dehydro-PGFz, (19) possesses luteo-lytic activity equal to that of PGF2, in a hamster antifertility assay.35In summary, the total synthesis of the prostacyclin analogue 1 is described, which mimicks natural PG12 in all respects so far examined, except for its 150 times greater half-life and its failure to be inactivated by the 15hydroxyprostaglandin dehydrogenase.The half-life of 1 was determined by incubating a (29) Hatano, Y. Department of Pharmacological and Physiological Sciences, University of Chicago, unpublished results. (30) The material prepared and tested in this work represents a 1:l mixture of diastereomers consisting of structure 1 and ita ent-15-epi derivative. To assess the potency of the latter, we prepared and examined ita nonfluorinated analogue for ita effect on the renal mesenteric artery. In contrast to 1, this substance caused contraction at the 10" M level, too high to substantially influence the effecta of 1. Our potency estimates are therefore reported in terms of a 50% content of 1. (31) Dusting, G. J.; Moncada, S.; Vane, J. R. Prostaglandins 1977, 13, 3. ( 3H)-pyrimidinones. Antiviral-and Interferon-Inducing AgentsSir: A survey of the variety of agents capable of stimulating interferon (IF) production demonstrates that most range in size from viruses and bacterial cell walls to synthetic or biopolymers [e.g., dextran and poly(I:C)].' However, there is also a small group of low-molecular-weight compounds which induce IF. Included among these compounds which have been rep...

2-Amino-5-bromo-6-methyl-4-pyrimidinol (U-25,166) induced high levels of circulating interferon in mice when administered either parenterally or orally. Peak titers of interferon were found in the serum between 6 and 12 h after inoculation of the drug. Lower but significant levels of interferon were found in rat serum after administration of U-25, 166 by either the intraperitoneal or oral route, and good levels of circulating interferon were observed in cats after oral treatment. Repeated intraperitoneal doses (50 mg/kg) of U-25, 166 protected mice against intranasal encephalomyocarditis virus challenge. The minimal effective acute oral dose for antiviral activity was approximately 250 mg/kg. This was also the minimal dose that produced detectable levels of interferon. Maximum tolerated doses in mice were four to six times the minimal effective doses. A single oral treatment was protective in mice against challenge virus inoculated 24 h later. The compound protected mice from challenge with high levels of encephalomyocarditis virus, up to 20,000 mean lethal doses. Antiviral activity in mice was retained when certain minor substitutions were made in the U-25,166 structure.

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