5T4-TARGETED IMMUNOFUSION MOLECULE AND METHODS
Immunofusion molecules useful for 5T4-targeted therapy. The immunofusion molecules include the 5T4 anti-gen-binding portion of an anti-5T4 antibody engineered into a single chain form and fused to a cytotoxic payload, such as, human pancreatic RNase (“HPRN”). The RNase portion of the single immunofusion peptide may be fused to a polyglutamic acid (polyE) tail. A pharmaceutical composition includes an immunofusion molecule including a 5T4 antigen-binding portion and HPRN and methods of administering the composition to an animal in need.
1 . An immunofusion molecule comprising an antigen-binding portion and an RNase portion in a single chain peptide, wherein:
a) the polypeptide sequence of the antigen-binding portion comprises SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:97 or SEQ ID NO:98, and
b) the polypeptide sequence of the RNase portion comprises any one of SEQ ID NO:13 to SEQ ID NO:20.
2 . The immunofusion molecule of claim 1 , wherein the antigen-binding portion and RNase portion are fused by a polypeptide linker.
3 . The immunofusion molecule of claim 2 , wherein the polypeptide linker has a polypeptide sequence according to any one of SEQ ID NO:9 to SEQ ID NO:12.
4 . The immunofusion molecule of claim 1 , further comprising a CathepsinB substrate sequence fused to N-terminus or C-terminus of the RNase portion.
5 - 6 . (canceled)
7 . The immunofusion molecule of claim 4 , further comprising at least ten glutamate residues fused to the C-terminus of the CathepsinB substrate sequence.
8 . (canceled)
9 . An immunofusion molecule comprising an antigen-binding portion and a RNase portion in a single-chain peptide, wherein the polypeptide sequence of the single-chain peptide comprises the polypeptide sequence according to any one of SEQ ID NO:32 to SEQ ID NO:95.
10 . The immunofusion molecule of claim 1 , wherein the single chain peptide further comprises an N-terminal signal sequence designated as SEQ ID NO:2.
11 . The immunofusion molecule of claim 1 , wherein the single-chain peptide forms a monomer of a dimer or a tetramer upon expression in a cell.
12 - 13 . (canceled)
14 . The immunofusion molecule of claim 9 , wherein the polypeptide sequence of the single-chain peptide is modified by insertion, substitution or deletion of one or more amino acids and the immunofusion molecule binds the 5T4-antigen.
15 . An isolated polynucleotide encoding a single-chain peptide comprising the polypeptide sequence according to any one of SEQ ID NO:32 to SEQ ID NO:95.
16 - 25 . (canceled)
26 . A method of treating a disease or disorder involving the expression of the 5T4 antigen comprising administering to an animal in need of such treatment a therapeutically effective amount of an immunofusion molecule comprising an antigen-binding portion and a RNase portion in a single chain peptide, wherein the polypeptide sequence of the antigen-binding portion comprises SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:97 or SEQ ID NO:98, and the polypeptide sequence of the RNase portion comprises any one of SEQ ID NO:13 to SEQ ID NO:20.
27 . The method of claim 26 , wherein the disease or disorder is cancer selected from the group consisting of colorectal cancer, bladder cancer, gastric cancer, breast cancer, lung cancer and prostate cancer.
28 - 32 . (canceled)
33 . An isolated polypeptide designated as SEQ ID NOs:21, 22, 97 or 98.
34 . An isolated polypeptide designated as SEQ ID NOs:101, 102 or 103.
35 . An immonufusion molecule comprising SEQ ID NO:103.