Heat Shock Protein-Binding Peptide Compositions And Methods Of Use Thereof

Abstract

Provided are polypeptides and compositions comprising novel HSP-binding peptides. Such polypeptides and compositions are particularly useful as immunotherapeutics (e.g., cancer vaccines). Also provided are methods of inducing a cellular immune response using such polypeptides and compositions, methods of treating a disease using such polypeptides and compositions, kits comprising such polypeptides and compositions, and methods of making such compositions.

1 .- 126 . (canceled) 
     
     
         127 . A composition comprising a complex of:
 (a) a polypeptide comprising:
 (i) an antigenic peptide comprising one or more major histocompatibility complex (MHC)-binding epitopes, and 
 (ii) an HSP-binding peptide comprising the amino acid sequence of SEQ ID NO: 6; and 
   (b) a purified stress protein.   
     
     
         128 . The composition of  claim 127 , wherein the amino acid sequence of the HSP-binding peptide consists of the amino acid sequence of SEQ ID NO: 6. 
     
     
         129 . The composition of  claim 127 , wherein the HSP-binding peptide is at the C-terminus of the polypeptide. 
     
     
         130 . The composition of  claim 127 , wherein the HSP-binding peptide is linked to the antigenic peptide via a peptide linker, optionally wherein the peptide linker comprises the amino acid sequence of SEQ ID NO: 13. 
     
     
         131 . The composition of  claim 127 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 18. 
     
     
         132 . The composition of  claim 127 , wherein the polypeptide is 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 amino acids in length. 
     
     
         133 . The composition of  claim 127 , wherein the MHC-binding epitope is from a cancer cell and wherein the MHC-binding epitope comprises an amino acid mutation or a gene fusion mutation, optionally wherein
 (a) the amino acid mutation is a substitution, deletion, or insertion mutation, and/or   (b) the amino acid mutation or gene fusion mutation is at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 of the amino acid sequence of the antigenic peptide.   
     
     
         134 . The composition of  claim 127 , wherein the MHC-binding epitope is from a pathogenic microbe, optionally wherein the pathogenic microbe is a virus. 
     
     
         135 . The composition of  claim 127 , wherein the MHC-binding epitope comprises a modified amino acid residue, optionally wherein:
 (a) the modified amino acid residue is a Tyr, Ser, Thr, Arg, Lys, or His that has been phosphorylated on a side chain hydroxyl or amine;   (b) the modified amino acid residue is a mimetic of a Tyr, Ser, Thr, Arg, Lys, or His amino acid that has been phosphorylated on a side chain hydroxyl or amine, optionally wherein the mimetic is a non-hydrolyzable analogue of a phosphorylated residue;   (c) the modified amino acid residue is an Asn that has been glycosylated on a side chain amide, a Ser or Thr that has been glycosylated on a side chain hydroxyl, a Lys or Arg that has been methylated on a side chain amino, a Lys that has been acetylated on a side chain amino, an N-terminal residue that has been acetylated on the α-amino, or a C-terminal residue that has been amidated on the α-carboxyl; and/or   (d) the modified amino acid residue is at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 of the amino acid sequence of the antigenic peptide.   
     
     
         136 . The composition of  claim 127 , wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 38-53. 
     
     
         137 . The composition of  claim 127 , wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 54-69. 
     
     
         138 . The composition of  claim 127 , wherein:
 (a) the stress protein is selected from the group consisting of Hsc70, Hsp70, Hsp90, Hsp110, Grp170, Gp96, Calreticulin, and a mutant or fusion protein thereof, optionally wherein the stress protein is human Hsc70;   (b) the stress protein is a recombinant protein;   (c) the composition comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 different complexes, optionally wherein each of the different complexes comprises the same HSP-binding peptide and a different antigenic peptide, optionally wherein:   
       each one of the polypeptides comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 54-69;
 (d) the amount of the stress protein in the composition is about 10 μg to 600 μg, optionally about 250 μg to 290 μg; 
 (e) the molar ratio of the polypeptide(s) to the stress protein is about 0.5:1 to 5:1, optionally about 1:1 to 2:1, such as 1:1, 1.25:1, or 1.5:1; 
 (f) the total amount of the polypeptide(s) and stress protein in the composition is about 10 μg to 600 μg, optionally about 300 μg; 
 (g) the composition further comprises an adjuvant, optionally wherein the adjuvant comprises:
 a saponin or an immunostimulatory nucleic acid, optionally wherein the adjuvant comprises QS-21, optionally about 10 μg, 25 μg, or 50 μg of the QS-21, and/or 
 a TLR agonist, optionally a TLR4 agonist, TLR5 agonist, TLR7 agonist, TLR8 agonist, and/or TLR9 agonist; 
 
 (h) the composition comprises a pharmaceutically acceptable carrier or excipient; and/or 
 (i) the composition is in a unit dosage form. 
 
     
     
         139 . The composition of  claim 127 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 18, and the stress protein is human Hsc70. 
     
     
         140 . The composition of  claim 139 , wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 38-53. 
     
     
         141 . The composition of  claim 139 , wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 54-69. 
     
     
         142 . The composition of  claim 139 , wherein:
 (a) the composition comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 different complexes, optionally wherein each of the different complexes comprise the same HSP-binding peptide and a different antigenic peptide, optionally wherein:   
       each one of the polypeptides comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 54-69;
 (b) the amount of the stress protein in the composition is about 10 μg to 600 μg, optionally about 250 μg to 290 μg; 
 (c) the molar ratio of the polypeptide(s) to the stress protein is about 0.5:1 to 5:1, optionally about 1:1 to 2:1, such as 1:1, 1.25:1, or 1.5:1; 
 (d) the total amount of the polypeptide(s) and stress protein in the composition is about 10 μg to 600 μg, optionally about 300 μg; 
 (e) the composition further comprises an adjuvant, optionally wherein the adjuvant comprises:
 a saponin or an immunostimulatory nucleic acid, optionally wherein the adjuvant comprises QS-21, optionally about 10 μg, 25 μg, or 50 μg of the QS-21, and/or 
 a TLR agonist, optionally a TLR4 agonist, TLR5 agonist, TLR7 agonist, TLR8 agonist, and/or TLR9 agonist; 
 
 (f) the composition comprises a pharmaceutically acceptable carrier or excipient; and/or 
 (g) the composition is in a unit dosage form. 
 
     
     
         143 . A method of making the composition of  claim 127 , the method comprising mixing the polypeptide with the purified stress protein under suitable conditions such that the purified stress protein binds to the polypeptide. 
     
     
         144 . A method making the composition of  claim 139 , the method comprising mixing the polypeptide with the purified stress protein under suitable conditions such that the purified stress protein binds to the polypeptide. 
     
     
         145 . A method of inducing a cellular immune response to an antigenic peptide in a subject, the method comprising administering to the subject an effective amount of the composition of  claim 127 . 
     
     
         146 . A method of inducing a cellular immune response to an antigenic peptide in a subject, the method comprising administering to the subject an effective amount of the composition of  claim 139 .