A library of the 8000 tripeptides derivable from coded amino acids was prepared in 20 sets of 400 using solid phase synthesis on a benzhydrylamine resin. The peptide mixtures, as C-terminal amides, were screened for inhibition of secreted alkaline phosphatase expression in a cellular (COS) system wherein a transfected SeAP gene construct was under control of the HIV-1 LTR promoter, activated by the product of a cotransfected HIV Tat gene construct. Thus, YPG-NH2 was discovered as an inhibitor of HIV-1 Tat function and then shown to block HIV replication in a CD4+ T-cell line (CEM) with IC50 = 35μM.
References
1.
BrayA.M.MaejiN.J., and GeysenH.M. (1990) The simultaneous multiple production of solution phase peptides; assessment of the Geysen method of simultaneous peptide synthesis. Tetrahedron Lett31: 5811–5814.
2.
DaytonA.I.SodroskiJ.G.RosenC.A.GohW.C., and HaseltineW.A. (1986) The frans-activator gene of the human T cell lymphotropic virus type III is required for replication. Cell44(6): 941–947.
3.
FisherA.G.FeinbergM.B.JosephsS.F.HarperM.E.MarselleL.M.ReyesG.GondaM.A.AldoviniA.DeboukC.GalloR.C., and Wong-StaalF. (1986) The frans-activator gene of HTLV-III is essential for virus replication. Nature320: 367–371.
4.
FodorS.P.A.ReadJ.L.PirrungM.C.StryerL.LuA.T., and SolasD. (1991) Light-directed spatially addressable parallel chemical synthesis. Science251: 767–773.
5.
HoughtenR.A.PinillaC.BlondelleS.E.AppelJ.R.DooleyC.T., and CuervoJ.H. (1991) Generation and use of synthetic peptide combinatorial libraries for basic research and drug discovery. Nature354: 84–86.
6.
HsuM.-C.SchuttA.D.HollyM.SliceL.W.ShermanM.I.RichmanD.D.PotashM.J., and VolskyD.J. (1991) Inhibition of HIV replication in acute and chronic infections in vitro by a TAT antagonist. Science254: 1799–1802.
7.
HsuM.-C.DhingraU.EarleyJ.V.HollyM.KeithD.NalinC.RichouA.R.SchuttA.D.TarnS.Y.PotashM.J.VolskyD.J., and RichmanD.D. (1993) Inhibition of Type 1 Human Immunodeficiency Virus replication by a Tat antagonist to which the virus remains sensitive after prolonged exposure in vitro. Proc Natl Acad Sci Usa90: 6395–6399.
LamK.S.SalmonS.E.HershE.M.HrubyV.J.KazmierskiW.M., and KnappR.J. (1991) A new type of synthetic peptide library for identifying ligand-binding activity. Nature354: 82–84.
10.
ZuckermannR.N.KerrJ.M.SianiM.A.BanvilleS.C., and SantiD.V. (1992) Identification of highest-affinity ligands by affinity selection from equimolar peptide mixtures generated by robotic synthesis. Proc Natl Acad Sci USA89: 4505–4509.
