Stabilized Soluble Pre-Fusion RSV F Polypeptides

Abstract

Described are stable pre-fusion respiratory syncitial virus (RSV) F polypeptides, immunogenic compositions comprising the polypeptides and uses thereof for the prevention and/or treatment of RSV infection.

1 . A vector comprising a nucleic acid molecule encoding a recombinant pre-fusion respiratory syncytial virus (RSV) Fusion (F) mutant polypeptide comprising a mutation of the amino acid residue N/T at position 67 into I and/or a mutation of the amino acid residue S at position 215 into P, wherein the amino acid positions are given in reference to the sequence of RSV F protein from the A2 strain (SEQ ID NO: 1), and the vector is an Ad26 vector or an Ad35 vector. 
     
     
         2 . The vector of  claim 1 , wherein the mutant polypeptide comprises at least one further mutation selected from the group consisting of:
 (a) a mutation of the amino acid residue on position 46;   (b) a mutation of the amino acid residue on position 77;   (c) a mutation of the amino acid residue on position 80;   (d) a mutation of the amino acid residue on position 92;   (e) a mutation of the amino acid residue on position 175;   (f) a mutation of the amino acid residue on position 184;   (g) a mutation of the amino acid residue on position 185;   (h) a mutation of the amino acid residue on position 201;   (i) a mutation of the amino acid residue on position 209;   (j) a mutation of the amino acid residue on position 421;   (k) a mutation of the amino acid residue on position 426;   (l) a mutation of the amino acid residue on position 465;   (m) a mutation of the amino acid residue on position 486;   (n) a mutation of the amino acid residue on position 487; and   (o) a mutation of the amino acid residue on position 508.   
     
     
         3 . The vector of  claim 2 , wherein the at least one further mutation is selected from the group consisting of:
 (a) a mutation of the amino acid residue S on position 46 into G;   (b) a mutation of the amino acid residue K on position 77 into E;   (c) a mutation of the amino acid residue K on position 80 into E;   (d) a mutation of the amino acid residue E on position 92 into D;   (e) a mutation of the amino acid residue N on position 175 into P;   (f) a mutation of the amino acid residue G on position 184 into N;   (g) a mutation of the amino acid residue V on position 185 into N;   (h) a mutation of the amino acid residue K on position 201 into Q;   (i) a mutation of the amino acid residue K on position 209 into Q;   (j) a mutation of the amino acid residue K on position 421 into N;   (k) a mutation of the amino acid residue N on position 426 into S;   (l) a mutation of the amino acid residue K on position 465 into E or Q;   (m) a mutation of the amino acid residue D on position 486 into N;   (n) a mutation of the amino acid residue E on position 487 into Q, N or I; and   (o) a mutation of the amino acid residue K on position 508 into E.   
     
     
         4 . The vector of  claim 1 , wherein the mutant polypeptide comprises a mutation of the amino acid residue N/T at position 67 into I and a mutation of the amino acid residue S at position 215 into P. 
     
     
         5 . A vector comprising a nucleic acid molecule encoding a recombinant pre-fusion respiratory syncytial virus (RSV) Fusion (F) mutant polypeptide, wherein the mutant polypeptide comprises a mutation of the amino acid residue N/T at position 67 into I, a mutation of the amino acid residue S at position 215 into P, a mutation of the amino acid residue K at position 66 into E, a mutation of the amino acid residue I at position 76 into V, and a mutation of the amino acid residue D at position 486 into N, wherein the amino acid positions are given in reference to the sequence of RSV F protein from the A2 strain (SEQ ID NO: 1), and the vector is an Ad26 vector or an Ad35 vector. 
     
     
         6 . The vector of  claim 5 , wherein the mutant polypeptide comprises a truncated F1 domain and an F2 domain. 
     
     
         7 . The vector of  claim 6 , wherein the mutant polypeptide comprises a heterologous trimerization domain linked to the truncated F1 domain. 
     
     
         8 . The vector of  claim 7 , wherein the heterologous trimerization domain comprises SEQ ID NO: 4. 
     
     
         9 . A vector comprising a nucleic acid molecule encoding a recombinant pre-fusion respiratory syncytial virus (RSV) Fusion (F) mutant polypeptide, wherein the mutant polypeptide comprises a mutation of the amino acid residue N/T at position 67 into I, a mutation of the amino acid residue S at position 215 into P, a mutation of the amino acid residue S at position 46 into G, a mutation of the amino acid residue K at position 66 into E, and a mutation of the amino acid residue I at position 76 into V, wherein the amino acid positions are given in reference to the sequence of RSV F protein from the A2 strain (SEQ ID NO: 1), and the vector is an Ad26 vector or an Ad35 vector. 
     
     
         10 . The vector of  claim 9 , wherein the mutant polypeptide comprises a truncated F1 domain and an F2 domain. 
     
     
         11 . The vector of  claim 10 , wherein the mutant polypeptide comprises a heterologous trimerization domain linked to the truncated F1 domain. 
     
     
         12 . The vector of  claim 11 , wherein the heterologous trimerization domain comprises SEQ ID NO: 4. 
     
     
         13 . A composition comprising the vector of  claim 1 , and a pharmaceutically acceptable carrier or excipient. 
     
     
         14 . A composition comprising the vector of  claim 5 , and a pharmaceutically acceptable carrier or excipient. 
     
     
         15 . A composition comprising the vector of  claim 8 , and a pharmaceutically acceptable carrier or excipient. 
     
     
         16 . A composition comprising the vector of  claim 9 , and a pharmaceutically acceptable carrier or excipient. 
     
     
         17 . A composition comprising the vector of  claim 12 , and a pharmaceutically acceptable carrier or excipient. 
     
     
         18 . A method of inducing an immune response against RSV F protein in a subject, the method comprising administering to the subject the vector of  claim 1  in an amount to induce an immune response against RSV F protein in the subject. 
     
     
         19 . A method of inducing an immune response against RSV F protein in a subject, the method comprising administering to the subject the vector of  claim 5  in an amount to induce an immune response against RSV F protein in the subject. 
     
     
         20 . A method of inducing an immune response against RSV F protein in a subject, the method comprising administering to the subject the vector of  claim 8  in an amount to induce an immune response against RSV F protein in the subject. 
     
     
         21 . A method of inducing an immune response against RSV F protein in a subject, the method comprising administering to the subject the vector of  claim 9  in an amount to induce an immune response against RSV F protein in the subject. 
     
     
         22 . A method of inducing an immune response against RSV F protein in a subject, the method comprising administering to the subject the vector of  claim 12  in an amount to induce an immune response against RSV F protein in the subject. 
     
     
         23 . A method of prophylaxing and/or treating RSV infection in a subject, the method comprising administering to the subject the vector of  claim 1 , to prophylax against and/or treat the subject for RSV infection. 
     
     
         24 . A method of prophylaxing and/or treating RSV infection in a subject, the method comprising administering to the subject the vector of  claim 5 , to prophylax against and/or treat the subject for RSV infection. 
     
     
         25 . A method of prophylaxing and/or treating RSV infection in a subject, the method comprising administering to the subject the vector of  claim 8 , to prophylax against and/or treat the subject for RSV infection. 
     
     
         26 . A method of prophylaxing and/or treating RSV infection in a subject, the method comprising administering to the subject the vector of  claim 9 , to prophylax against and/or treat the subject for RSV infection. 
     
     
         27 . A method of prophylaxing and/or treating RSV infection in a subject, the method comprising administering to the subject the vector of  claim 12 , to prophylax against and/or treat the subject for RSV infection.