Mutant interleukin-2 polypeptides

Patent metadata

Field	Value
Publication number	US-2016208017-A1
Application number	US-201614996789-A
Country	US
Kind code	A1
Filing date	Jan 15, 2016
Priority date	Feb 10, 2011
Publication date	Jul 21, 2016
Grant date	—

Abstract

The present invention generally relates to mutant interleukin-2 polypeptides that exhibit reduced affinity to the α-subunit of the IL-2 receptor, for use as immunotherapeutic agents. In addition, the invention relates to immunoconjugates comprising said mutant IL-2 polypeptides, polynucleotide molecules encoding the mutant IL-2 polypeptides or immunoconjugates, and vectors and host cells comprising such polynucleotide molecules. The invention further relates to methods for producing the mutant IL-2 polypeptides or immunoconjugates, pharmaceutical compositions comprising the same, and uses thereof.

First claim

1 . An immunoconjugate comprising a mutant IL-2 polypeptide and an antigen binding moiety, wherein said mutant IL-2 polypeptide is a human IL-2 molecule comprising the amino acid substitutions F42A, Y45A and L72G according to the numbering relative to the human IL-2 sequence SEQ ID NO: 1; and
wherein said antigen binding moiety is an IgG 1 subclass immunoglobulin molecule specific for FAP, comprising (i) the heavy chain variable region sequence of SEQ ID NO: 41 and the light chain variable region sequence of SEQ ID NO: 39; (ii) the heavy chain variable region sequence of SEQ ID NO: 51 and the light chain variable region sequence of SEQ ID NO: 49; (iii) the heavy chain variable region sequence of SEQ ID NO: 111 and the light chain variable region sequence of SEQ ID NO: 109; (iv) the heavy chain variable region sequence of SEQ ID NO: 143 and the light chain variable region sequence of SEQ ID NO: 141; or (iv) the heavy chain variable region sequence of SEQ ID NO: 151 and the light chain variable region sequence of SEQ ID NO: 149.

2 . The immunoconjugate of claim 1 , wherein said mutant IL-2 polypeptide further comprises the amino acid substitution T3A and/or the amino acid substitution C125A.

3 . The immunoconjugate of claim 1 , wherein said mutant IL-2 polypeptide comprises the sequence of SEQ ID NO: 19.

4 . The immunoconjugate of claim 1 , wherein said mutant IL-2 polypeptide is joined at its amino-terminal amino acid to the carboxy-terminal amino acid of one of the immunoglobulin heavy chains.

5 . The immunoconjugate of claim 1 , wherein the immunoconjugate comprises not more than one mutant IL-2 polypeptide.

6 . The immunoconjugate of claim 1 , wherein the IgG 1 subclass immunoglobulin molecule comprises in the Fc domain a modification promoting heterodimerization of two non-identical immunoglobulin heavy chains.

7 . The immunoconjugate of claim 6 , wherein the modification is a knob-into-hole modification, comprising a knob modification in one of the immunoglobulin heavy chains and a hole modification in the other one of the immunoglobulin heavy chains.

8 . The immunoconjugate of claim 7 , wherein the knob modification comprises the amino acid substitution T366W in one of the two immunoglobulin heavy chains, and the hole modification comprises the amino acid substitutions T366S, L368A and Y407V in the other one of the two immunoglobulin heavy chains according to the EU numbering of Kabat.

9 . The immunoconjugate of claim 8 , wherein the immunoglobulin heavy chain comprising the knob modification additionally comprises the amino acid substitution S354C, and the immunoglobulin heavy chain comprising the hole modification additionally comprises the amino acid substitution Y349C according to the EU numbering of Kabat.

10 . The immunoconjugate of claim 1 , wherein the IgG 1 molecule comprises in its Fc domain one or more amino acid mutation that reduces the binding affinity of the immunoconjugate to an FcγRIIIa, FcγRI or FcγRIIa receptor.

11 . The immunoconjugate of claim 10 , wherein the IgG 1 molecule comprises the amino acid mutations L234A, L235A and P329G according to the EU numbering of Kabat.

12 . The immunoconjugate of claim 1 , comprising (i) a polypeptide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 297, a polypeptide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 299, and a polypeptide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 233; (ii) a polypeptide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 301, a polypeptide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 303, and a polypeptide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 231; or (iii) a polypeptide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 315, a polypeptide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 317, and a polypeptide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 233.

13 . The immunoconjugate of claim 1 , wherein said IgG 1 molecule specific for FAP comprises the heavy chain variable region sequence of SEQ ID NO: 111 and the light chain variable region sequence of SEQ ID NO: 109.

14 . The immunoconjugate of claim 1 , comprising a polypeptide sequence of SEQ ID NO: 301, a polypeptide sequence of SEQ ID NO: 303, and a polypeptide sequence of SEQ ID NO: 231.

15 . The immunoconjugate of claim 1 , consisting essentially of a mutant IL-2 polypeptide and an IgG 1 molecule, joined by a linker sequence.

16 . An isolated polynucleotide encoding the immunoconjugate of claim 1 .

17 . A host cell comprising the polynucleotide of claim 16 .

18 . A method of producing an immunoconjugate comprising a mutant IL-2 polypeptide and an antigen binding moiety, comprising culturing the host cell of claim 17 under conditions suitable for the expression of the immunoconjugate.

19 . An immunoconjugate produced by the method of claim 18 .

20 . A pharmaceutical composition comprising the immunoconjugate of claim 14 and a pharmaceutically acceptable carrier.

21 . A method of treating disease in an individual, comprising administering to said individual a therapeutically effective amount of a composition comprising the immunoconjugate of claim 14 in a pharmaceutically acceptable form.

22 . The method of claim 21 , wherein said disease is cancer.

23 . A method of stimulating the immune system of an individual, comprising administering to said individual an effective amount of a composition comprising the immunoconjugate of claim 14 in a pharmaceutically acceptable form.

Assignees

• Roche Glycart Ag

Inventors

• Ast Oliver
• Bruenker Peter
• Freimoser-Grundschober Anne
• Herter Sylvia
• Hofer Thomas U
• Hosse Ralf
• Klein Christian
• Moessner Ekkehard
• Nicolini Valeria G
• Umana Pablo

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P37/04

  Immunostimulants · CPC title

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title

• A61K47/6849

  the antibody targeting a receptor, a cell surface antigen or a cell surface determinant · CPC title

• C07K16/40 Primary

  against enzymes · CPC title