Cleavable vaccine compositions and uses thereof and methods of making and using the same
View Patent ↗The invention relates to methods for making vaccines using linkages that are cleavable under lysosomal processing conditions, and vaccine compositions obtained therefrom. In some embodiments, the vaccines comprise a tumor antigen, an immunogenic carrier and a linker covalently linking the tumor antigen and the immunogenic carrier by a thio ether linkage. Vaccines of preferred embodiments can be used against a cellular proliferative disease that is characterized by the tumor antigen.
1. A method of making a cancer vaccine which comprises
conjugating an immunogenic carrier comprising keyhole limpet hemocyanin (KLH) with a partially-reduced tumor-specific idiotypic immunoglobulin using a linker comprising a first reactive moiety which is maleimide and a second reactive moiety under conditions which permit the formation of a thio-ether bond between the partially-reduced tumor-specific idiotypic immunoglobulin and the first reactive moiety and a covalent bond between the immunogenic carrier and the second reactive moiety to give the cancer vaccine,
wherein the thio-ether bond between the linker and the partially-reduced tumor-specific idiotypic immunoglobulin is cleavable under lysosomal processing conditions, and
wherein the tumor-specific idiotypic immunoglobulin is associated with a B-cell proliferative disease selected from the group consisting of B-cell lymphoma, leukemia, multiple myeloma and light chain amyloidosis.
2. The method according to claim 1 , wherein the partially-reduced tumor-specific idiotypic immunoglobulin is obtained using a mild reducing agent selected from the group consisting of dithiothrietol (DTT), 2-mercaptoethanol, and 2-mercaptoethylamine.
3. The method according to claim 1 , wherein the partially-reduced tumor-specific idiotypic immunoglobulin is about 50% reduced.
4. The method according to claim 1 , wherein the partially-reduced tumor-specific idiotypic immunoglobulin is provided in a solution comprising phosphate-buffered saline and EDTA.
5. The method according to claim 1 , wherein the tumor-specific idiotypic immunoglobulin is from a malignant B cell.
6. The method according to claim 5 , wherein the tumor-specific idiotypic immunoglobulin is derived from an idiotype-producing hybridoma.
7. The method according to claim 5 , wherein the tumor-specific idiotypic immunoglobulin is recombinantly generated using DNA.
8. The method according to claim 1 , wherein the efficacy of the vaccine is independent of the duration of the conjugation step.
9. The cancer vaccine made by the method of claim 1 .
10. A cancer vaccine comprising a partially-reduced tumor-specific idiotypic immunoglobulin conjugated to an immunogenic carrier comprising keyhole limpet hemocyanin (KLH) by a bifunctional linker having at least one thio-ether bond between a maleimide of the bifunctional linker and the immunogenic carrier or the partially-reduced tumor-specific idiotypic immunoglobulin, wherein the thio-ether bond is cleavable under lysosomal processing conditions, and
wherein the tumor-specific idiotypic immunoglobulin is associated with a B-cell proliferative disease selected from the group consisting of B-cell lymphoma, leukemia, multiple myeloma and light chain amyloidosis.
11. A method of treating a cancer in a patient which comprises
administering to the patient a therapeutic amount the cancer vaccine according to claim 10 , wherein the cancer is a B-cell proliferative disease.
12. The method according to claim 11 , further comprising administering to the patient simultaneously or sequentially an immunologic adjuvant.
13. The method according to claim 12 , wherein the adjuvant is selected from the group consisting of GM-CSF, a toll-like receptor (TLR) agonist, poly I:C, and CpG DNA.
14. The method according to claim 11 , wherein the cancer expresses one or more tumor-specific idiotypic immunoglobulins.
15. A cancer vaccine which comprises
a tumor-specific idiotypic immunoglobulin;
an immunogenic carrier comprising keyhole limpet hemocyanin (KLH);
a bifunctional linker having a first moiety which is maleimide and a second moiety;
wherein the tumor-specific idiotypic immunoglobulin is conjugated to the immunogenic carrier by the bifunctional linker, and
wherein the first moiety forms a thio-ether bond with the tumor-specific idiotypic immunoglobulin or the immunogenic carrier, said thio-ether bond is cleavable under lysosomal processing conditions, and
wherein the tumor-specific idiotypic immunoglobulin is associated with a B-cell proliferative disease selected from the group consisting of B-cell lymphoma, leukemia, multiple myeloma and light chain amyloidosis.
16. The cancer vaccine of claim 15 , wherein the cancer vaccine is effective against a B cell lymphoma in a subject when administered to the subject.