Cytotoxic conjugates comprising a chemokine receptor targeting agent

• US 7,166,702 B1
• Filed: 12/02/1999
• Issued: 01/23/2007
• Est. Priority Date: 07/22/1998
• Status: Expired due to Fees

• Current Assignee: Osprey Pharmaceuticals USA, Inc. (Osprey Pharmaceuticals Limited)
• Original Assignee: Osprey Pharmaceuticals Limited
• Inventors: Coggins, Philip J., McDonald, John R.
• Primary Examiner(s): Landsman; Robert S.
• Application Number:
  US09/453,851
• Time in Patent Office: 2,609 Days
• Field of Search: 530/402, 530/387.1, 514/2
• US Class Current: 530/351
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: A61K 47/642 the peptide or protein in the drug conjugate being a cytokine, e.g. IL2, chemokine, growth factors or interferons being the inactive part of the conjugate
• CPC Class Codes: A61K 47/6425 the peptide or protein in the drug conjugate being a receptor, e.g. CD4, a cell surface antigen, i.e. not a peptide ligand targeting the antigen, or a cell surface determinant, i.e. a part of the surface of a cell
• CPC Class Codes: A61K 47/6811 the drug being a protein or peptide, e.g. transferrin or bleomycin
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C07K 14/521 Chemokines
• CPC Class Codes: C07K 2318/20 Antigen-binding scaffold molecules wherein the scaffold is not an immunoglobulin variable region or antibody mimetics
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/01 containing a localisation/targetting motif
• CPC Class Codes: C07K 2319/55 containing a fusion with a toxin, e.g. diphteria toxin
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids

Abstract

Conjugates containing as a ligand chemokine receptor targeting agents, such as chemokines, and a targeted agent, such as a toxin are provided. These conjugates are used to treat inflammatory responses associated with activation, proliferation and migration of immune effector cells, including leukocyte cell types, neutrophils, macrophages, and eosinophils. The conjugates provided herein are used to lessen or inhibit these processes to prevent or at least lessen the resulting secondary effects. In particular, the conjugates can be used to target toxins to receptors on secondary tissue damage-promoting cells. The ligand moiety can be selected to deliver the cell toxin to such secondary tissue damage-promoting cells as mononuclear phagocytes, leukocytes, natural killer cells, dendritic cells, and T and B lymphocytes, thereby suppressing the proliferation, migration, or physiological activity of such cells. Among preferred conjugates are fusion proteins having a chemokine, or a biologically active fragment thereof, as the ligand moiety linked to a cell toxin via a peptide linker of from 2 to about 60 amino acid residues.

Claims

1. A conjugate, comprising a targeted agent and a chemokine receptor targeting agent, or a portion thereof, wherein the conjugate binds to a chemokine receptor present on activated leukocytes resulting in internalization of the targeted agent in cells bearing the receptor.

2. The conjugate of claim 1, wherein the leukocytes are selected basophils, neutrophils, eosinophils, and combinations of any two or more thereof.

3. The conjugate of claim 1, wherein the chemokine and targeted agent are linked directly.

4. The conjugate of claim 1, wherein the chemokine and targeted agent are joined via a linker.

5. The conjugate of claim 4, wherein the linker comprises a peptide or a polypeptide or is a chemical linker.

6. The conjugate of claim 4, wherein the linker is a peptide or an amino acid.

7. A conjugate of claim 1, wherein the chemokine is selected from the group consisting of eotaxin, stromal derived factor-1β

, SDF-1α

, monocyte chemotactic protein-1 (MCP-1), MCP-3, macrophage inhibitory protein 1α

(MIP-1α

), MIP-1β and

Regulated on Activation, Normal T cell Expressed and Secreted (RANTES) protein.

8. A conjugate of claim 1 selected from among OPL98104, OPL98112, OPL98108, OPL98102, OPL98110, OPL98106, OPL98101, OPL98109, OPL98105, OPL98103, OPL98111 and OPL98107.

9. The conjugate of claim 1, wherein the chemokine effects binding to a receptor selected from the group consisting of CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, XCR1 and CX3CR-1, whereby the conjugate is internalized into a cell bearing the receptor.

10. The conjugate of claim 1, wherein the targeted agent is a toxin, and the toxin is a ribosome inactivating protein, a bacterial toxin or a fragment thereof that retains the ribosome inactivating or toxin activity.

11. The conjugate of claim 4, wherein the linker is selected from the group consisting of N-succinimidyl(4-iodoacetyl)-aminobenzoate, sulfosuccinimydil(4-iodoacetyl)-aminobenzoate, 4-succinimidyl-oxycarbonyl-α

-(2-pyridyldithio)toluene, sulfosuccinimidyl-6-(α

-methyl-α

-(pyridyldithiol)-toluamido) hexanoate, N-succinimidyl-3-(-2-pyridyldithio)-proprionate, succinimidyl6(3(-(-2-pyridyldithio)-proprionamido)hexanoate, sulfosuccinimidyl6(3(-(-2-pyridyldithio)-propionamido) hexanoate, 3-(2-pyridyldithio)-propionyl hydrazide, Ellman'"'"'s reagent, dichlorotriazinic acid, and S-(2-thiopyridyl)-L-cysteine.

12. The conjugate of claim 1, wherein the targeted agent is a toxin.

13. The conjugate of claim 12, wherein the toxin is a polypeptide.

14. A conjugate, comprising a targeted agent and a chemokine receptor targeting agent, or a portion thereof, wherein the conjugate binds to a chemokine receptor present on immune cells resulting in internalization of the targeted agent in cells bearing the receptor, wherein:

the chemokine receptor is selected from among chemokine receptors expressed on immune cells to which IL 8, GCP 2, GRP γ

, ENA 78, PBP, CTAP III, NAP 2, LAPF 4, MIG, IP 10, SDF 1α

, SDF 1β

, SDF 2, MCP 1, MCP 2, MCP 3, MCP 4, MCP 5, MIP 1α

, MIP 1β

, MIP 1γ

, MIP 2, MIP 2α

, MIP 3α

, MIP 3β

, MIP 4, MIP 5, MDC, HCC 1, eotaxin 1, eotaxin 2, I 309, SCYA17, Tim 1, TARC, RANTES, DC CK 1, lymphotactin, Tim 1, ALP, lungkine or fractalkine bind; and

the chemokine is selected from among IL 8, GCP 2, GRP γ

, ENA 78, PBP, CTAP III, NAP 2, LAPF 4, MIG, IP 10, SDF 1α

, SDF 1β

, SDF 2, MCP 1, MCP 2, MCP 3, MCP 4, MCP 5, MIP 1α

, MIP 1β

, MIP 1γ

, MIP 2, MIP 2α

, MIP 3α

, MIP 3β

, MIP 4, MIP 5, MDC, HCC 1, eotaxin 1, eotaxin 2, I 309, SCYA17, Tim 1, TARC, Regulated on Activation, Normal T cell Expressed and Secreted (RANTES) protein (RANTES), DC CK 1, lymphotactin, Tim 1, ALP, lungkine and fractalkine.

15. The conjugate of claim 14, wherein the chemokine is selected from among eotaxin-1, eotaxin-2, MCP-1, MCP-3, SDF-1β

and RANTES.

16. The conjugate of claim 14, wherein the targeted agent is a toxin that, upon internalization into a cell, alters metabolism or gene expression in the cell, regulates or alters protein synthesis, inhibits proliferation or kills he cell.

17. The conjugate of claim 15, wherein the targeted agent is a toxin that, upon internalization into a cell, alters metabolism or gene expression in the cell, regulates or alters protein synthesis, inhibits proliferation or kills he cell.