Polypeptides comprising immunoglobulin single variable domains targeting Glypican-3 and T cell receptor
The present technology aims at providing a novel type of drug for treating a subject suffering from cancer. Specifically, the technology provides polypeptides comprising at least four immunoglobulin single variable domains (ISVDs), characterized in that one ISVD binds to TCR and at least two ISVDs bind to GPC3. The present technology also provides nucleic acids, vectors and compositions.
1 . A polypeptide that comprises an immunoglobulin single variable domain (ISVD) that specifically binds GPC3, wherein said ISVD comprises three complementarity determining regions (CDR1 to CDR3, respectively), and wherein the ISVD comprises:
a) a CDR1 that comprises the amino acid sequence of SEQ ID NO: 7; a CDR2 that comprises the amino acid sequence of SEQ ID NO: 11 or an amino acid sequence with 1 amino acid difference with SEQ ID NO: 11, wherein the amino acid difference occurs at the fifth amino acid of SEQ ID NO: 11, and wherein any amino acid difference that occurs at the fifth amino acid of SEQ ID NO: 11 is selected from the group consisting of a substitution of lysine with glycine and a substitution of lysine with alanine; and a CDR3 that comprises the amino acid sequence of SEQ ID NO: 15,
b) a CDR1 that comprises the amino acid sequence of SEQ ID NO: 8; a CDR2 that comprises the amino acid sequence of SEQ ID NO: 12 or an amino acid sequence with 2 or 1 amino acid difference(s) with SEQ ID NO: 12, wherein the amino acid difference that occurs at the sixth amino acid of SEQ ID NO: 12 is a substitution of glutamate with aspartate, and wherein the amino acid difference that occurs at the ninth amino acid of SEQ ID NO: 12 is a substitution of tyrosine with asparagine; and a CDR3 that comprises the amino acid sequence of GVA,
c) a CDR1 that comprises the amino acid sequence of SEQ ID NO: 32; a CDR2 that comprises the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence with 2 or 1 amino acid difference(s) with SEQ ID NO: 36, wherein the amino acid difference that occurs at the fifth amino acid of SEQ ID NO: 36 is selected from the group consisting of a substitution of lysine with glycine and a substitution of lysine with alanine, and wherein the amino acid difference that occurs at the eleventh amino acid of SEQ ID NO: 36 is a substitution of alanine with arginine; and a CDR3 that comprises the amino acid sequence of SEQ ID NO: 15, or
d) a CDR1 that comprises the amino acid sequence of SEQ ID NO: 33; a CDR2 that comprises the amino acid sequence of SEQ ID NO: 37 or an amino acid sequence with 2 or 1 amino acid difference(s) with SEQ ID NO: 37, wherein the amino acid difference that occurs at the sixth amino acid of SEQ ID NO: 37 is a substitution of glutamate with aspartate, and wherein the amino acid difference that occurs at the ninth amino acid of SEQ ID NO: 37 is a substitution of tyrosine with asparagine; and a CDR3 that comprises the amino acid sequence of GVA.
2 . The polypeptide according to claim 1 , wherein said polypeptide further comprises one or more other groups, residues, moieties or binding units, in which said one or more other groups, residues, moieties or binding units provide the polypeptide with increased half-life, compared to the corresponding polypeptide without said one or more other groups, residues, moieties or binding units.
3 . The polypeptide according to claim 1 , in which said binding unit that provides the polypeptide with increased half-life is an ISVD that binds to human serum albumin, wherein said ISVD that binds to human serum albumin comprises;
a CDR1 that comprises the amino acid sequence of SEQ ID NO: 9, a CDR2 that comprises the amino acid sequence of SEQ ID NO: 13, and a CDR3 that comprises the amino acid sequence of SEQ ID NO: 17; or
a CDR1 that comprises the amino acid sequence of SEQ ID NO: 34, a CDR2 that comprises the amino acid sequence of SEQ ID NO: 38, and a CDR3 that comprises the amino acid sequence of SEQ ID NO: 17.
4 . The polypeptide according to claim 3 , wherein the amino acid sequence of said ISVD binding to human serum albumin exhibits a sequence identity of more than 90% with SEQ ID NO: 5, or wherein said ISVD binding to human serum albumin consists of the amino acid sequence of SEQ ID NO: 5.
5 . A nucleic acid comprising a nucleotide sequence that encodes a polypeptide according to claim 1 .
6 . A host or host cell comprising a nucleic acid according to claim 5 .
7 . A method for producing a polypeptide, said method comprising the step of expressing, in a suitable host cell or host organism or in another suitable expression system, a nucleic acid according to claim 5 .
8 . A composition comprising at least one polypeptide according to claim 1 .
9 . The composition according to claim 8 , which is a pharmaceutical composition which further comprises at least one pharmaceutically acceptable carrier, diluent or excipient and/or adjuvant.
10 . A method of treating cancer, wherein said method comprises administering, to a subject in need thereof, a pharmaceutically active amount of a polypeptide according to claim 1 , wherein the cancer is a liver cancer or a lung cancer.