Method of developing a vaccine using peptide-poly IC complexes
The invention describes the development of more potent peptide vaccines to prevent or treat infections or cancer. Small synthetic peptides from the known sequences of viral, bacterial, parasitic or tumor antigens are modified so they can spontaneously form complexes with a synthetic nucleic acid, such as Poly IC, that functions as an immunological adjuvant. The peptide-nucleic acid complexes are dramatically more immunogenic as compared to the separate components. The procedure for developing the vaccine involves the conjugation of a synthetic peptide containing a C residue to poly-K using a bi-functional cross-linking reagent (SMCC). The peptide/poly-K complex was then formulated with CMC and poly-IC to produce a self-adjuvant vaccine that was 36-fold more effective as compared to the same peptide administered mixed with the same adjuvant (but not complexed to it).
1. A vaccine comprising:
an immune adjuvant wherein the immune adjuvant is a synthetic nucleic acid inducing a cell-mediated response and selected from the group consisting of polyinosinic:polycytidylic acid (Poly-IC), Poly-IC/LC, Poly-K, Poly-IC12U, and carboxymethyl cellulose (CMC); and
a modified peptide comprising:
a T cell epitope for infectious agents or tumor antigens wherein the T cell epitope is between 8 to 15 amino acids in length;
a short hydrophobic amino acid linker; and
at least one arginine (R) residue wherein the at least one arginine residue is appended to an amino or a carboxy terminal end of the T-cell epitope;
wherein the modified peptide consists of the T-cell epitope, the short hydrophobic amino acid linker, and the at least one arginine residue without any additional amino acids;
wherein interaction of the immune adjuvant and the modified peptide form a complex.
2. The vaccine of claim 1 , wherein the at least one arginine residue is added to the amino terminal end of the T-cell epitope.
3. The vaccine of claim 1 , wherein between about 3 to about 10 of the arginine residues are added to the T-cell epitope.
4. The vaccine of claim 1 , wherein the immune adjuvant is complexed to the modified peptide via covalent bonds.
5. The vaccine of claim 1 , wherein a chemical cross-linking reagent is used to complex the immune adjuvant to the at least one modified peptide.
6. A vaccine comprising:
an immune adjuvant wherein the immune adjuvant is selected from the group consisting of polyinosinic:polycytidylic acid (Poly-IC), Poly-IC/LC, Poly-K, Poly-IC12U, and carboxymethyl cellulose (CMC); and
a modified peptide comprising:
a known T cell epitope for infectious agents or tumor antigens wherein the T cell epitope is between 8 to 15 amino acids in length; and
between about 3 to about 20 hydrophobic residues wherein the hydrophobic residues are appended to an amino or a carboxy end of the I-cell epitope;
wherein the hydrophobic residues are selected from the group consisting of hydrophobic residues having aromatic rings, small linear hydrophobic amino acid sequences having at least 6 amino acid residues, and lipids;
wherein interaction of the immune adjuvant and the modified peptide form a complex.
7. The vaccine of claim 6 , wherein the hydrophobic residues are aromatic rings selected from the group consisting of phenylalanine (F) and tryptophan (W).
8. The vaccine of claim 6 , wherein the hydrophobic residues are added to the amino terminal end of the T-cell epitope.
9. The vaccine of claim 6 , further comprising the modified peptide being further modified by the addition of at least 1 arginine (R) residue.
10. A vaccine comprising:
an immune adjuvant wherein the immune adjuvant is selected from the group consisting of polyinosinic:polycytidylic acid (Poly-IC), Poly-IC/LC, and Poly-IC12U; and
a modified peptide comprising:
a known T cell epitope for infectious agents or tumor antigens wherein the T cell epitope is between 8 to 15 amino acids in length;
at least one hydrophilic amino acid linker; and
a cysteine (C) residue wherein the cysteine residue is appended to an amino or a carboxy end of the T-cell epitope;
wherein the modified peptide consists of the T-cell epitope, the at least one hydrophilic amino acid linker and the cysteine residue without any additional amino acids;
wherein the modified peptide is conjugated via covalent bonds to a polymer selected from the group consisting of Poly-K, Poly-IC and carboxymethyl cellulose (CMC) using a chemical cross-linking reagent (CLR) to form a conjugate;
wherein interaction of the immune adjuvant and the conjugate form a complex.
11. The vaccine of claim 10 , wherein a chemical cross-linking reagent (CLR) is used to complex the immune adjuvant to the at least one modified peptide.
12. The vaccine of claim 11 , wherein the CLR is succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC).
13. The vaccine of claim 10 , wherein the cysteine residue is added to the amino terminal end of the T-cell epitope.