Therapeutic anticancer neoepitope vaccine
The present invention relates to an anticancer vaccine which includes polynucleotides or polypeptides, methods of treatment of cancer wherein such an anticancer vaccine is used as well as methods for producing the vaccine. The vaccine includes a polynucleotide with a nucleotide sequence encoding a targeting unit, a dimerization unit, a first linker and an antigenic unit. The antigenic unit includes from 3 to 50 antigenic subunits separated by a second linker with each antigenic subunit having at least a part of a cancer neoepitope sequence. The vaccine can include a polypeptide encoded by the polynucleotide or a dimeric protein with two polypeptides encoded by the polynucleotide.
1 . An anticancer neoepitope composition comprising:
a DNA polynucleotide encoding a polypeptide comprising:
a targeting unit,
a dimerization unit,
a first linker, and
an antigenic unit having n cancer neoepitope sequences, wherein the antigenic unit is comprised of n−1 antigenic subunits, each subunit comprising a cancer neoepitope sequence having a length of 7 to 30 amino acids and a second linker; wherein n is an integer of from 10 to 20.
2 . The anticancer neoepitope composition according to claim 1 , wherein the antigenic unit comprises one copy of each of n different cancer neoepitope sequences.
3 . The anticancer neoepitope composition according to claim 1 , wherein the antigenic unit comprises at least two copies of at least one cancer neoepitope sequence.
4 . The anticancer neoepitope composition according to claim 1 , wherein each cancer neoepitope sequence has identical length.
5 . The anticancer neoepitope composition according to claim 1 , wherein a cancer neoepitope is positioned essentially in a middle of each cancer neoepitope sequence.
6 . The anticancer neoepitope composition according to claim 1 , wherein the antigenic subunits are in an order of more antigenic to less antigenic from the first linker toward a final antigenic subunit.
7 . The anticancer neoepitope composition according to claim 1 , wherein the antigenic subunit(s) comprising the most hydrophobic cancer neoepitope sequences is/are substantially the middle of the antigenic unit and the antigenic subunit(s) comprising the most hydrophilic cancer neoepitope sequences is/are at the ends of the antigenic unit.
8 . The anticancer neoepitope composition according to claim 1 , wherein the second linker is a flexible linker.
9 . The anticancer neoepitope composition according to claim 1 , wherein the second linker is a Serine-Glycine linker.
10 . The anticancer neoepitope composition according to claim 1 , wherein the length of the antigenic unit is from about 300 amino acids to about 1000 amino acids.
11 . The anticancer neoepitope composition according to claim 1 , wherein the cancer neoepitope sequence is from a cancer resulting in a tumor, such as a cancer having a high mutational load, and/or a cancer selected from the group consisting of melanoma, lung cancer, breast cancer, prostate cancer and colon cancer.
12 . The anticancer neoepitope composition according to claim 1 , wherein the DNA polynucleotide is comprised in a vector.
13 . The anticancer neoepitope composition according to claim 1 , wherein the DNA polynucleotide further encodes a signal peptide.
14 . The anticancer neoepitope composition according to claim 1 , wherein the targeting unit: a) comprises antibody binding regions with specificity for cluster of differentiation 14 (CD14), cluster of differentiation 40 (CD40), Toll-like receptor such as Toll-like receptor 2 (TLR-2), Toll-like receptor 4 (TLR-4), and Toll-like receptor (TLR-5), or major histocompatibility complex (MHC) class II protein; or b) is a ligand selected from the group consisting of soluble CD40 ligand, chemokines, such as regulated on activation, normal T-cell expressed and secreted (RANTES) or macrophage inflammatory protein 1α (MIP-1a) and a bacterial antigen, such as flagellin.
15 . The anticancer neoepitope composition according to claim 1 , wherein the dimerization unit comprises a hinge region and another domain that facilitates dimerization, wherein the hinge region and the other domain are connected through a third linker.
16 . The anticancer neoepitope composition according to claim 1 , wherein the polypeptide comprises, in N-terminal to C-terminal order, the targeting unit, the dimerization unit and the antigenic unit.
17 . A method of preparing the anticancer neoepitope composition according to claim 1 , wherein the method comprises a) identifying the neoepitopes to be encoded by the DNA polynucleotide and b) preparing the DNA polynucleotide.
18 . The method according to claim 17 , wherein the neoepitopes in a) are identified by a method comprising i) sequencing the genome or exome of a patient's tumor, ii) identifying tumor neoantigens comprising neoepitopes from said tumor; and iii) selecting neoepitopes based on predicted antigenicity.
19 . A method of treating a cancer in a patient, the method comprising administering to the patient the anticancer neoepitope composition of claim 1 .