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Arenaviruses and filoviruses are capable of causing hemorrhagic fever syndrome in humans. Limited therapeutic and/or prophylactic options are available for humans suffering from viral hemorrhagic fever. In this report, we demonstrate that pre-treatment of host cells with the kinase inhibitors genistein and tyrphostin AG1478 leads to inhibition of infection or transduction in cells infected with Ebola virus, Marburg virus, and Lassa virus. In all, the results demonstrate that a kinase inhibitor cocktail consisting of genistein and tyrphostin AG1478 is a broad-spectrum antiviral that may be used as a therapeutic or prophylactic against arenavirus and filovirus hemorrhagic fever.

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The vaccinia virus-encoded heterodimer responsible for poly(A) tail elongation comprises a polyadenylylation catalytic subunit (VP55) and associated processivity factor (VP39). We show that monomeric VP39's affinity for RNA homopolymers follows the hierarchy poly(I) >poly(U) >>poly(G) >poly(A) >poly(C), that the heterodimer interacts stably with 40-45 nucleotide nucleic acid segments, and that its homopolymer preference for polyadenylylation priming is comparable to the VP39 affinity hierarchy (above). For oligonucleotide ligands possessing the previously-identified (rU)2-(N)25-rU heterodimer-binding motif, the heterodimer's affinity and base-type preference are mediated via both the (rU)2and rU portions, with the greater contribution coming from (rU)2. VP39's R107 sidechain contributes to specificity at the downstream rU. Substitution of each ribouridylate of the motif with either ribothymidine or 4-thiodeoxythymidine indicated that the downstream rU interacts with both heterodimer subunits, whereas the upstream (rU)2interacts only with VP55. A 'crosslinking SELEX' approach indicated VP39-base proximity around position -10 of a 4-thioribouridine/deoxycytidine ligand pool. Upon incubating the heterodimer with a panel of identical-sequence oligonucleotides derivatized with azidophenacyl bromide at various phosphate positions, those derivatized at positions -11 to -21 photocrosslinked to both subunits in a coordinated manner. This region may therefore pass through a 'cleft' or enclosed 'channel' at the subunit interface.

Irradiation of Purines 6187 filtrates concentrated by vacuum distillation through a 14" column. The residue is subjected to vacuum distillation through a 5" column. The desired aminoalcohol distils as a viscous oil, b.p. 104-106°( 0.1 mm.), yield 56.3 g., (90.8%), re26D 1.5087. 4-Morpholino-2-butynyl N-Methylpipecolinate.-Into a 500-cc. 3-necked round bottom flask equipped with stirrer, reflux condenser, Dean-Stark water separator (CaCla tube) and heating mantle is placed a solution of 31.4 g. (0.20 mole) of methyl N-methylpipecolinate and 31.0 g. (0.20 mole) of 4-morpholino-2-butyn-l-ol in 325 cc. of re-heptane; 0.5 g. of NaOMe is added and the mixture refluxed. The methanol produced during the transesterification will separate from the heptane in the water separator. Two additional 0.3-g. portions of NaOMe may be required to complete the reaction. The reaction mixture is concentrated by slowly dis-tilling off approximately 50% of the heptane. The residue is chilled, filtered and the balance of the heptane removed by vacuum distillation through a 14" column. The residue is subjected to vacuum distillation through a 3" column. The desired ester boils at 149-151°( 0.25 mm.), yield 41.6 g. (74.3%), re26D 1.5012.4-Morpholino-2-butynyl N-Methylpipecolinate Dimethobromide.-To a solution of 14.0 g. (0.05 mole) of 4-morpholino-2-butynyl-N-methylpipecolinate (0.05 mole) in 80 cc. of isopropyl alcohol is added 19.0 g. (0.20 mole) of methyl bromide. The mixture is refluxed under anhydrous conditions for 3 hr. and then chilled. The solid is filtered off and recrystallized from the minimum amount of hot ethanol, m.p. 208-210°dec., yield 20.8 g. (88.5%).