Immune-stimulating monoclonal antibodies against human interleukin-2
The invention relates to a human Interleukin-2 (hIL-2) specific monoclonal antibody (mAb), or antigen binding fragment thereof, the binding of which to hIL-2 inhibits binding of hIL-2 to CD25 and the antibody is characterized by any of the parameters: the variable chain of the mAb comprises the amino acid sequence of SEQ ID NO 005 or SEQ ID NO 006; the binding to hIL-2 is characterized by a dissociation constant (K D )≤7.5 nmol/L; the binding to hIL-2 is characterized by an off-rate (K off )≤1×10 −4 s −1 and/or the antibody displays no measurable cross-reactivity to murine IL-2.
1. A human interleukin-2 (hIL-2) specific monoclonal antibody (mAb), or a hIL-2 specific antigen binding fragment thereof comprising a heavy chain complementarity determining region (HCDR) HCDR1 according to SEQ ID NO: 7, HCDR2 according to SEQ ID NO: 8, HCDR3 according to SEQ ID NO: 9, a light chain complementarity determining region (LCDR) LCDR1 according to SEQ ID NO: 10, LCDR2 according to SEQ ID NO: 11, LCDR3 according to SEQ ID NO: 12, wherein binding of said antibody, or antigen binding fragment thereof, to hIL-2 inhibits binding of hIL-2 to CD25.
2. The hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 1 , characterized in that it comprises at least one heavy chain variable region (VH) and/or one light chain variable region (VL) sequence having a sequence identity of ≥90% compared to SEQ ID NOs 019 or 20, and wherein the hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, comprises the HCDR1 according to SEQ ID NO: 7, the HCDR2 according to SEQ ID NO: 8, the HCDR3 according to SEQ ID NO: 9, the LCDR1 according to SEQ ID NO: 10, the LCDR2 according to SEQ ID NO: 11, the LCDR3 according to SEQ ID NO: 12.
3. The hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 1 , wherein the antibody, or antigen binding fragment thereof, displays no measurable cross-reactivity to murine IL-2.
4. The antibody or antigen binding fragment thereof of claim 1 , which binds to a hIL-2 epitope comprising the amino acids K52, P54, K55, T57, R58, T61, F62, K63, Q94, and K96of the amino acid sequence of SEQ ID NO:1.
5. The antibody or antigen binding fragment thereof according to claim 4 , wherein the epitope further comprises any one or more of the amino acids N50, N53, N91, L92, A93, N97 of the amino acid sequence of SEQ ID NO:1.
6. The hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 1 , comprising at least one VH and/or at least one VL sequence, wherein said VH and/or VL sequence comprises SEQ ID NOs 019 or 20.
7. The hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 1 , comprising an amino acid sequence of having a sequence identity of ≥90% to SEQ ID NOs 005 or 006, and wherein the hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, comprises the HCDR1 according to SEQ ID NO: 7, the HCDR2 according to SEQ ID NO: 8, the HCDR3 according to SEQ ID NO: 9, the LCDR1 according to SEQ ID NO: 10, the LCDR2 according to SEQ ID NO: 11, the LCDR3 according to SEQ ID NO: 12.
8. The hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 1 , comprising at least one amino acid sequence of SEQ ID NOs 005 or 006.
9. A therapeutic formulation comprising the hIL-2 monoclonal antibody (mAb), or the antigen binding fragment thereof, according to claim 1 and human interleukin-2 and a pharmaceutically acceptable carrier.
10. A therapeutic formulation comprising a fusion protein and a pharmaceutically acceptable carrier, wherein said fusion protein comprises:
a. an hIL-2 binding polypeptide fragment comprising HCDR1 according to SEQ ID NO: 7, HCDR2 according to SEQ ID NO: 8, HCDR3 according to SEQ ID NO: 9, LCDR1 according to SEQ ID NO: 10, LCDR2 according to SEQ ID NO: 11, LCDR3 according to SEQ ID NO: 12, wherein the hIL-2 binding polypeptide fragment binds to hIL-2 and inhibits the binding of hIL-2 to CD25,
b. a human IL-2 polypeptide fragment having an identity of ≥90% compared to SEQ ID NO 001, and, optionally,
c. an amino acid linker of 1 to 50 amino acids, linking the hIL-2 binding polypeptide fragment to the human IL-2 polypeptide fragment as one single polypeptide chain.
11. The therapeutic formulation of claim 10 , wherein the amino acid linker is 5 to 40 amino acids in length.
12. The therapeutic formulation of claim 10 , wherein the amino acid linker is 10 to 30 amino acids in length.
13. The therapeutic formulation of claim 10 , wherein the amino acid linker is 15 to 25 amino acids in length.
14. The therapeutic formulation of claim 10 , wherein the hIL-2 binding polypeptide fragment is further characterized as:
a. comprising an amino acid sequence having an identity of ≥90% compared to SEQ ID NO: 19 or SEO ID NO: 20, wherein said hIL-2 binding polypeptide fragment comprises the HCDR1 according to SEQ ID NO: 7, the HCDR2 according to SEQ ID NO: 8, the HCDR3 according to SEQ ID NO: 9, the LCDR1 according to SEQ ID NO: 10, the LCDR2 according to SEQ ID NO: 11, the LCDR3 according to SEQ ID NO: 12; and/or
b. displaying no measurable cross reactivity to murine IL-2.
15. A nucleic acid molecule encoding the hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 1 .
16. The nucleic acid molecule of claim 15 , wherein said nucleic acid molecule has ≥90% sequence identity compared to SEQ ID NOs 003 or 004 and wherein said nucleic acid encodes the HCDR1 according to SEQ ID NO: 7, the HCDR2 according to SEQ ID NO: 8, the HCDR3 according to SEQ ID NO: 9, the LCDR1 according to SEQ ID NO: 10, the LCDR2 according to SEQ ID NO: 11, the LCDR3 according to SEQ ID NO: 12.
17. The nucleic acid molecule according to claim 15 , wherein said nucleic acid molecule encodes the hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 2 .
18. The nucleic acid molecule according to claim 15 , wherein said nucleic acid molecule encodes the hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 6 .
19. The nucleic acid molecule according to claim 15 , wherein said nucleic acid molecule encodes the hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 7 .
20. The nucleic acid molecule according to claim 15 , wherein said nucleic acid molecule encodes the hIL-2 specific monoclonal antibody, or antigen binding fragment thereof, according to claim 8 .
21. A vector comprising the nucleic acid molecule of claim 15 .
22. A cell comprising the nucleic acid molecule according to claim 15 or expressing the nucleic acid molecule according to claim 15 .
23. A cell expressing hIL-2 specific monoclonal antibody (mAb), or antigen binding fragment thereof, according to claim 1 .
24. A monoclonal antibody-producing hybridoma cell line that produces the hIL-2 specific antibody, or antibody fragment thereof, of claim 1 .
25. A method of treating cancer or viral infection by (1) selecting a patient having cancer or viral infection and (2) administering a therapeutically effective amount of the antibody, or antigen binding fragment thereof of claim 1 , in combination with a therapeutically effective amount of human interleukin-2.