De-immunized Shiga toxin a subunit effector polypeptides for applications in mammals

A polynucleotide encoding a cell-targeting molecule comprising a protein, wherein the protein consists of:

• (a) a binding region capable of specifically binding an extracellular target biomolecule physically coupled to the surface of a cell, wherein the extracellular target biomolecule is not CD38;
• (b) a Shiga toxin effector polypeptide comprising an amino acid sequence having at least 90% identity to amino acids 1 to 251 of SEQ ID NO:
• 1,
• wherein the amino acid sequence comprises at least four endogenous B-cell epitope regions, and further comprises:
• i) a plurality of disrupted endogenous B-cell epitope regions, wherein the disrupted endogenous B-cell epitope regions contain the following amino acid substitutions:
• S45 of SEQ ID NO:
• 1 to I, R55 of SEQ ID NO:
• 1 to L, 157 of SEQ ID NO:
• 1 to F, P59 of SEQ ID NO:
• 1 to F, E60 of SEQ ID NO:
• 1 to T, E61 of SEQ ID NO:
• 1 to L, G110 of SEQ ID NO:
• 1 to A, R188 of SEQ ID NO:
• 1 to A, R248 of SEQ ID NO:
• 1 to A and R251 of SEQ ID NO:
• 1 to A;
• and ii) the amino acid substitution C242 of SEQ ID NO:
• 1 to S;
• and
• wherein the amino acid sequence comprises an asparagine at the amino acid residue corresponding to position 75 of SEQ ID NO:
• 1, a tyrosine at the amino acid residue corresponding to position 77 of SEQ ID NO:
• 1, a tyrosine at the amino acid residue corresponding to position 114 of SEQ ID NO:
• 1, a glutamate at the amino acid residue corresponding to position 167 of SEQ ID NO:
• 1, an arginine at the amino acid residue corresponding to position 170 of SEQ ID NO:
• 1, an arginine at the amino acid residue corresponding to position 176 of SEQ ID NO:
• 1, and a tryptophan at the amino acid residue corresponding to position 203 of SEQ ID NO:
• 1;
• and
• (c) a linker between the binding region and the Shiga toxin effector polypeptide.