Bispecific binding agents for modulating biological activity
Methods for improving the biological and pharmaceutical properties of bispecific binding agents are described herein where the bispecific binding agent are able to target cells by a high affinity binding domain to a first cell surface marker that does not induce a significant biological effect and a low affinity binding domain that binds specifically to a second cell surface marker, causing a significant and desired biological effect. Compositions of such bispecific binding agents, uses for them, and kits containing them are also provided.
1. A method for treating a malignancy comprising administering to a patient an effective amount of a bispecific binding agent capable of modulating biological activity of target cancer cells that have a first and a second target antigen on their exterior surface, wherein
(i) said first and second target-antigens do not share a common ligand,
(ii) said second target antigen is a receptor tyrosine kinase, and
(iii) said bispecific binding agent has a first binding domain that is an antibody that binds to the first target antigen with a dissociation constant (Kd) for the first target antigen of 10 −7 M or less and a second binding domain that is an antibody that binds to the second target antigen with a Kd for the second target antigen that is at least 10 times greater than the Kd of the first binding domain, and
when the first target antigen of said first binding domain is ErbB2 (HER2), the second target antigen for said second binding domain is not ErbB3 (HER3); and,
wherein said binding of said first antigen to said first binding domain does not modulate biological activity of said first target antigen, but said binding of said second binding domain to said second target antigen modulates the biological activity of said second target antigen, and the biological activity results in either an inhibition of target cell proliferation or death of the target cell.
2. A method of claim 1 , wherein the antibodies are diabodies, single chain Fvs, disulfide stabilized Fvs, or combinations thereof, and are connected directly or by a linker.
3. A method of claim 1 , wherein the second target antigen is a tumor-associated antigen, cytokine receptor, or growth factor receptor.
4. A method of claim 1 , wherein said first target antigen is selected from the group consisting of carcinoembryonic antigen (CEA), ErbB2, EGFR, Lewis Y , MUC-1, EpCAM, CA125, prostate specific membrane antigen (PSMA), and TAG72.
5. A method of claim 1 , wherein the second target antigen is selected from the group consisting of ErbB3, ErbB4 , any of FGF receptors 1-4, HGF receptor, IGF1-R, PDGF receptors alpha and beta, and C-KIT.
6. A method of claim 1 , wherein the Kd of the first binding domain for the first target antigen is between 10 −8 M and 10 −12 M.
7. A method of claim 1 , wherein the Kd of the second binding domain to the second target antigen is at least 20 times greater than the Kd of the first binding domain to the first target antigen.
8. A method of claim 1 , wherein the Kd of the second binding domain to the second target antigen is at least 50 times greater than the Kd of the first binding domain to the first target antigen.
9. A method of claim 1 wherein the patient is a human patient, and the target cell is a cell in the patient.