Long lasting fusion peptide inhibitors of viral infection

Abstract

Peptides exhibiting anti-viral and anti-fusogenic activity are modified to provide greater stability and improved half-life in vivo. The selected peptides include fusion inhibitors DP178 and DP107 and related peptides and analogs thereof. The modified peptides are capable of forming covalent bonds with one or more blood components, preferably a mobile blood component.

1 . A modified anti-viral peptide comprising:
 a peptide that exhibits anti-viral activity against respiratory syncytial virus (RSV), and   a reactive group which is reactive with amino groups, hydroxyl groups, or thiol groups on blood components to form stable covalent bonds.   
     
     
         2 . The modified peptide of  claim 1 , wherein said reactive group is a succinimidyl or a maleimido group. 
     
     
         3 . The modified peptide of  claim 1 , wherein said reactive group is a maleimido group which is reactive with a thiol group on a blood protein. 
     
     
         4 .- 8 . (canceled) 
     
     
         9 . The modified peptide of  claim 1 , wherein said peptide is selected from the group consisting of SEQ ID NO: 10 to SEQ ID NO: 30. 
     
     
         10 . The modified peptide of  claim 1 , wherein said peptide is selected from the group consisting of SEQ ID N0: 14 to SEQ ID N0: 17 and SEQ ID NO: 29. 
     
     
         11 . A modified anti-viral peptide comprising:
 a peptide exhibits antiviral activity against human parainfluenza virus (HPIV), and   a reactive group which is reactive with amino groups, hydroxyl groups, or thiol groups on blood components to form stable covalent bonds.   
     
     
         12 . The modified peptide of  claim 11 , wherein said peptide is selected from the group consisting of SEQ ID N0: 31 to SEQ ID N0: 62. 
     
     
         13 . The modified peptide of  claim 11 , wherein said peptide is selected from the group consisting of SEQ ID NO: 35, SEQ ID N0: 38 to SEQ ID N0: 42, SEQ ID N0: 52 and SEQ ID N0: 58. 
     
     
         14 . A modified anti-viral peptide comprising:
 a peptide exhibits antiviral activity against measles virus (MeV), and   a reactive group which is reactive with amino groups, hydroxyl groups, or thiol groups on blood components to form stable covalent bonds.   
     
     
         15 . The modified peptide of  claim 14 , wherein said peptide is selected from the group consisting of SEQ ID N0: 74 to SEQ ID N0: 86. 
     
     
         16 . The modified peptide of  claim 14 , wherein said peptide is selected from the group consisting of SEQ ID N0: 77, SEQ ID N0: 79, SEQ ID N0: 81 and SEQ ID N0: 84. 
     
     
         17 . A modified anti-viral peptide comprising:
 a peptide exhibits antiviral activity against simian immunodeficiency virus (SIV), and   a reactive group which is reactive with amino groups, hydroxyl groups, or thiol groups on blood components to form stable covalent bonds.   
     
     
         18 . The modified peptide of  claim 17 , wherein said peptide is selected from the group consisting of SEQ ID N0: 63 to SEQ ID N0: 73. 
     
     
         19 . (canceled) 
     
     
         20 . The modified peptide of  claim 11 , wherein said reactive group is a maleimido group which is reactive with a thiol group on a blood protein. 
     
     
         21 .- 22 . (canceled) 
     
     
         23 . The modified peptide of  claim 14 , wherein said reactive group is a maleimido group which is reactive with a thiol group on a blood protein. 
     
     
         24 .- 25 . (canceled) 
     
     
         26 . The modified peptide of  claim 17 , wherein said reactive group is a maleimido group which is reactive with a thiol group on a blood protein. 
     
     
         27 .- 30 . (canceled) 
     
     
         31 . The modified peptide of  claim 11 , wherein said reactive group is a succinimidyl or a maleimido group. 
     
     
         32 . The modified peptide of  claim 14 , wherein said reactive group is a succinimidyl or a maleimido group. 
     
     
         33 . The modified peptide of  claim 17 , wherein said reactive group is a succinimidyl or a maleimido group. 
     
     
         34 . An anti-viral peptide-albumin conjugate comprising:
 a peptide that exhibits anti-viral activity against respiratory syncytial virus (RSV) that comprises an amino acid sequence and a reactive group which is reactive with amino groups, hydroxyl groups, or thiol groups on blood components to form stable covalent bonds; and   albumin, wherein the peptide is covalently bonded to the albumin through the reactive group.   
     
     
         35 . The conjugate of  claim 34 , wherein said reactive group is a maleimido group which is reactive with a thiol group on the albumin. 
     
     
         36 . The conjugate of  claim 34 , wherein said peptide is selected from the group consisting of SEQ ID NO: 10 to SEQ ID NO: 30. 
     
     
         37 . The conjugate of  claim 34 , wherein said peptide is selected from the group consisting of SEQ ID N0: 14 to SEQ ID N0: 17 and SEQ ID NO: 29. 
     
     
         38 . An anti-viral peptide-albumin conjugate comprising:
 a peptide that exhibits anti-viral activity against human parainfluenza virus (HPIV), that comprises an amino acid sequence and a reactive group which is reactive with amino groups, hydroxyl groups, or thiol groups on blood components to form stable covalent bonds; and   albumin, wherein the peptide is covalently bonded to the albumin through the reactive group.   
     
     
         39 . The conjugate of  claim 38 , wherein said reactive group is a maleimido group which is reactive with a thiol group on the albumin. 
     
     
         40 . The conjugate of  claim 38 , wherein said peptide is selected from the group consisting of SEQ ID N0: 31 to SEQ ID N0: 62. 
     
     
         41 . The conjugate of  claim 38 , wherein said peptide is selected from the group consisting of SEQ ID N0: 35, SEQ ID N0: 38 to 42, SEQ ID N0: 52 and SEQ ID N0: 58. 
     
     
         42 . An anti-viral peptide-albumin conjugate comprising:
 a peptide that exhibits anti-viral activity against measles virus (MeV), that comprises an amino acid sequence and a reactive group which is reactive with amino groups, hydroxyl groups, or thiol groups on blood components to form stable covalent bonds; and   albumin, wherein the peptide is covalently bonded to the albumin through the reactive group.   
     
     
         43 . The conjugate of  claim 42 , wherein said reactive group is a maleimido group which is reactive with a thiol group on the albumin. 
     
     
         44 . The conjugate of  claim 42 , wherein said peptide is selected from the group consisting of SEQ ID N0: 74 to SEQ ID N0: 86. 
     
     
         45 . The conjugate of  claim 42  wherein said peptide is selected from the group consisting of SEQ ID N0: 77, SEQ ID N0: 79, SEQ ID N0: 81 and SEQ ID N0: 84. 
     
     
         46 . An anti-viral peptide-albumin conjugate comprising:
 a peptide that exhibits anti-viral activity against simian immunodeficiency virus (SIV), that comprises an amino acid sequence and a reactive group which is reactive with amino groups, hydroxyl groups, or thiol groups on blood components to form stable covalent bonds; and   albumin, wherein the peptide is covalently bonded to the albumin through the reactive group.   
     
     
         47 . The conjugate of  claim 46 , wherein said reactive group is a maleimido group which is reactive with a thiol group on the albumin. 
     
     
         48 . The conjugate of  claim 46 , wherein said peptide is selected from the group consisting of SEQ ID N0: 63 to SEQ ID N0: 73. 
     
     
         49 . A composition comprising the modified anti-viral peptide of  claim 1  and a physiologically acceptable medium. 
     
     
         50 . A composition comprising the modified anti-viral peptide of  claim 11  and a physiologically acceptable medium. 
     
     
         51 . A composition comprising the modified anti-viral peptide of  claim 14  and a physiologically acceptable medium. 
     
     
         52 . A composition comprising the modified anti-viral peptide of  claim 17  and a physiologically acceptable medium. 
     
     
         53 . A composition comprising the conjugate of  claim 34  and a physiologically acceptable medium. 
     
     
         54 . A composition comprising the conjugate of  claim 38  and a physiologically acceptable medium. 
     
     
         55 . A composition comprising the conjugate of  claim 42  and a physiologically acceptable medium. 
     
     
         56 . A composition comprising the conjugate of  claim 46  and a physiologically acceptable medium. 
     
     
         57 . A method of inhibiting or reducing membrane fusion between respiratory syncytial virus (RSV) and a cell, comprising contacting the RSV with a modified peptide of  claim 1 . 
     
     
         58 . A method of inhibiting or reducing membrane fusion between human parainfluenze virus (HPIV) and a cell, comprising contacting the HPIV with a modified peptide of  claim 11 . 
     
     
         59 . A method of inhibiting or reducing membrane fusion between measles virus (MeV) and a cell, comprising contacting the MeV with a modified peptide of  claim 14 . 
     
     
         60 . A method of inhibiting or reducing membrane fusion between simian immunodeficiency virus (SIV) and a cell, comprising contacting the SIV with a modified peptide of  claim 17 . 
     
     
         61 . A method of inhibiting or reducing membrane fusion between respiratory syncytial virus (RSV) and a cell, comprising contacting the RSV with a conjugate of  claim 34 . 
     
     
         62 . A method of inhibiting or reducing membrane fusion between human parainfluenze virus (HPIV) and a cell, comprising contacting the HPIV with a conjugate of  claim 38 . 
     
     
         63 . A method of inhibiting or reducing membrane fusion between measles virus (MeV) and a cell, comprising contacting the MeV with a conjugate of  claim 42 . 
     
     
         64 . A method of inhibiting or reducing membrane fusion between simian immunodeficiency virus (SIV) and a cell, comprising contacting the SIV with a conjugate of  claim 46 . 
     
     
         65 . A method of making a conjugate of  claim 34 , comprising:
 contacting a modified peptide of  claim 1  with albumin to form an anti-viral peptide-albumin conjugate.   
     
     
         66 . The method of  claim 65 , wherein the modified anti-viral peptide is made by combining an anti-RSV peptide having a free amino group, N-[γ-maleimidobutyryloxy]succinimide ester (GMBS) and trietylamine or maleimidopropionic acid (MPA) to form a maleimide containing group at the free amino group, to form the modified anti-viral peptide. 
     
     
         67 . A method of making a conjugate of  claim 38 , comprising:
 contacting a modified peptide of  claim 11  with albumin to form an anti-viral peptide-albumin conjugate.   
     
     
         68 . The method of  claim 67 , wherein the modified anti-viral peptide is made by combining an anti-HPIV peptide having a free amino group, N-[γ-maleimidobutyryloxy]succinimide ester (GMBS) and trietylamine or maleimidopropionic acid (MPA) to form a maleimide containing group at the free amino group, to form the modified anti-viral peptide. 
     
     
         69 . A method of making a conjugate of  claim 42 , comprising:
 contacting a modified peptide of  claim 14  with albumin to form an anti-viral peptide-albumin conjugate.   
     
     
         70 . The method of  claim 69 , wherein the modified anti-viral peptide is made by combining an anti-MeV peptide having a free amino group, N-[γ-maleimidobutyryloxy]succinimide ester (GMBS) and trietylamine or maleimidopropionic acid (MPA) to form a maleimide containing group at the free amino group, to form the modified anti-viral peptide. 
     
     
         71 . A method of making a conjugate of  claim 46 , comprising:
 contacting a modified peptide of  claim 17  with albumin to form an anti-viral peptide-albumin conjugate.   
     
     
         72 . The method of  claim 71 , wherein the modified anti-viral peptide is made by combining an anti-SIV peptide having a free amino group, N-[γ-maleimidobutyryloxy]succinimide ester (GMBS) and trietylamine or maleimidopropionic acid (MPA) to form a maleimide containing group at the free amino group, to form the modified anti-viral peptide. 
     
     
         73 . A method of treating or preventing respiratory syncytial virus (RSV) infection in a subject, comprising
 administering a modified peptide of  claim 1  to a subject having or at risk of RSV infection, thereby treating or preventing the infection.   
     
     
         74 . The method of  claim 73 , wherein the subject has brochiolitis. 
     
     
         75 . The method of  claim 73 , wherein the subject has pneumonia. 
     
     
         76 . The method of  claim 73 , wherein the modified peptide is conjugated with albumin in vivo. 
     
     
         77 . The method of  claim 73 , wherein the modified peptide is conjugated with albumin ex vivo. 
     
     
         78 . A method of treating or preventing respiratory syncytial virus (RSV) infection in a subject, comprising
 administering a conjugate of  claim 34  to a subject having or at risk of RSV infection, thereby treating or preventing the infection.   
     
     
         79 . The method of  claim 78 , wherein the subject has brochiolitis. 
     
     
         80 . The method of  claim 78 , wherein the subject has pneumonia. 
     
     
         81 . A method of treating or preventing human parainfluenza virus (HPIV) infection in a subject, comprising
 administering a modified peptide of  claim 11  to a subject having or at risk of HPIV infection, thereby treating or preventing the infection.   
     
     
         82 . The method of  claim 81 , wherein the subject has respiratory tract disease. 
     
     
         83 . The method of  claim 81 , wherein the subject has croup. 
     
     
         84 . The method of  claim 81 , wherein the subject has brochiolitis. 
     
     
         85 . The method of  claim 81 , wherein the subject has pneumonia. 
     
     
         86 . The method of  claim 81 , wherein the modified peptide is conjugated with albumin in vivo. 
     
     
         87 . The method of  claim 81 , wherein the modified peptide is conjugated with albumin ex vivo. 
     
     
         88 . A method of treating or preventing human parainfluenza virus (HPIV) infection in a subject, comprising
 administering a conjugate of  claim 38  to a subject having or at risk of HPIV infection, thereby treating or preventing the infection.   
     
     
         89 . The method of  claim 88 , wherein the subject has respiratory tract disease. 
     
     
         90 . The method of  claim 88 , wherein the subject has croup. 
     
     
         91 . The method of  claim 88 , wherein the subject has brochiolitis. 
     
     
         92 . The method of  claim 88 , wherein the subject has pneumonia. 
     
     
         93 . A method of treating or preventing measles virus (MeV) infection in a subject, comprising
 administering a modified peptide of  claim 14  to a subject having or at risk of MeV infection, thereby treating or preventing the infection.   
     
     
         94 . The method of  claim 93 , wherein the modified peptide is conjugated with albumin in vivo. 
     
     
         95 . The method of  claim 93 , wherein the modified peptide is conjugated with albumin ex vivo. 
     
     
         96 . A method of treating or preventing measles virus (MeV) infection in a subject, comprising
 administering a conjugate of  claim 42  to a subject having or at risk of MeV infection, thereby treating or preventing the infection.   
     
     
         97 . A method of treating or preventing simian immunodeficiency virus (SIV) infection in a subject, comprising
 administering a modified peptide of  claim 17  to a subject having or at risk of SIV infection, thereby treating or preventing the infection.   
     
     
         98 . The method of  claim 97 , wherein the modified peptide is conjugated with albumin in vivo. 
     
     
         99 . The method of  claim 97 , wherein the modified peptide is conjugated with albumin ex vivo. 
     
     
         100 . A method of treating or preventing simian immunodeficiency virus (SIV) infection in a subject, comprising
 administering a conjugate of  claim 46  to a subject having or at risk of SIV infection, thereby treating or preventing the infection.