Enhanced immunogenicity of tumor associated antigens by addition of alphagal epitopes
The invention relates to methods and compositions for causing the selective targeting and killing of tumor cells. The present invention describes prophylactic or therapeutic cancer vaccines based on purified TAA proteins or TAA-derived synthetic peptides altered by chemical, enzymatic or chemo-enzymatic methods to introduce αGal epitopes or αGal glycomimetic epitopes, in order to allow for enhanced opsonization of the antigen by natural anti-αGal antibodies to stimulate TAA capture and presentation, thereby inducing a humoral and cellular immune response to the TAA expressed by a tumor. The animal's immune system thus is stimulated to produce tumor specific cytotoxic cells and antibodies which will attack and kill tumor cells present in the animal.
1. An antitumor vaccine composition effective in a patient comprising: a purified and isolated tumor-associated antigen (TAA) protein or peptide, said TAA protein or peptide comprising a sequence of 7-20 contiguous amino acids comprised in the amino acid sequence of a TAA protein and at least one flanking sequence comprising one or more amino acids selected from the group consisting of: lysine, cysteine, homocysteine, serine, threonine, and glutamine, said TAA protein or peptide further having two or more αGal epitopes covalently bound to said flanking sequence by means of a cross-linking agent.
2. The antitumor vaccine of claim 1 , wherein the TAA protein or peptide comprises an affinity purification tag.
3. The antitumor vaccine of claim 1 wherein the TAA protein or peptide is synthetic.
4. The antitumor vaccine of claim 1 , wherein said TAA is expressed by said patient to be treated.
5. An antitumor vaccine composition effective in a patient comprising: a TAA-derived peptide comprising a sequence of 7-20 contiguous amino acids comprised in the amino acid sequence of a TAA protein, and further comprising at least one flanking sequence comprising one or more amino acids selected from the group consisting of: lysine, cysteine, homocysteine, serine, threonine, and glutamine, and further having two or more αGal epitopes covalently bound to said flanking sequence, by means of a cross-linking agent.
6. The antitumor vaccine of claim 5 , wherein the peptide is of a length of 6-50 amino acids.
7. The antitumor vaccine of claim 5 , wherein the TAA is expressed by said patient to be treated.
8. The antitumor vaccine of claim 1 , wherein the two or more αGal epitopes are independently trisaccharides of formula Galα1-3Galβ1-4Glc, or Galα1-3Galβ1-4GlcNAc, wherein R 1 is any linear or branched alkyl group of 1 to 30 carbon atoms, wherein one or more carbon atoms in said alkyl group can be substituted by O, S, or N and wherein one or more hydrogens can be substituted by hydroxyl, carbonyl, alkyl, sulphydryl or amino groups, and wherein R 2 is an amino or sulphydryl reacting group, and a carrier.
9. An antitumor vaccine composition effective in a patient comprising: a TAA-derived peptide comprising an amino acid sequence consisting of the amino acid sequence of any one of SEQ ID NOs: 20-74 and at least one flanking sequence comprising one or more amino acids selected from the group consisting of: lysine, cysteine, homocysteine, serine, threonine, and glutamine, said TAA protein or peptide further having two or more αGal epitopes covalently bound to said flanking sequence by means of a cross-linking agent.