Compositions and methods for targeted immunomodulatory antibodies and fusion proteins

• US 8,993,524 B2
• Filed: 03/04/2011
• Issued: 03/31/2015
• Est. Priority Date: 03/05/2010
• Status: Active Grant

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Bedi, Atul, Ravi, Rajani
• Primary Examiner(s): Shafer, Shulamith H
• Application Number:
  US13/582,717
• Publication Number:
  US 20130039911A1
• Time in Patent Office: 1,488 Days
• Field of Search: None
• US Class Current: 514/19.2
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/572 cytotoxic response
• CPC Class Codes: A61K 31/704 attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin digitoxin A61K31/7048
• CPC Class Codes: A61K 38/179 for growth factors; for growth regulators
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/495 Transforming growth factor [TGF]
• CPC Class Codes: C07K 14/70532 B7 molecules, e.g. CD80, CD86
• CPC Class Codes: C07K 14/70575 NGF/TNF-superfamily, e.g. CD70, CD95L, CD153, CD154 NGF C07K14/48, TNF C07K14/525
• CPC Class Codes: C07K 14/70578 NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 14/70596 Molecules with a "CD"-designation not provided for elsewhere
• CPC Class Codes: C07K 14/71 : for growth factors; for growth regulators
• CPC Class Codes: C07K 16/28 : against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2812 : against CD4
• CPC Class Codes: C07K 16/2815 : against CD8
• CPC Class Codes: C07K 16/2827 : against B7 molecules, e.g. CD80, CD86
• CPC Class Codes: C07K 16/2851 : against the lectin superfamily, e.g. CD23, CD72
• CPC Class Codes: C07K 16/2863 : against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/2866 : against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/30 : from tumour cells
• CPC Class Codes: C07K 19/00 : Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: C07K 2317/33 : Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/526 : CH3 domain
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction

Abstract

The present invention is based on the seminal discovery that targeted immunomodulatory antibodies and fusion proteins can counter act or reverse immune tolerance of cancer cells. Cancer cells are able to escape elimination by chemotherapeutic agents or tumor-targeted antibodies via specific immunosuppressive mechanisms in the tumor microenvironment and such ability of cancer cells is recognized as immune tolerance. Such immune suppressive mechanisms include immunosuppressive cytokines (for example, Transforming growth factor beta (TGF-β) and regulatory T cells and/or immunosuppressive myeloid dendritic cells (DCs). By counteracting tumor-induced immune tolerance, the present invention provides effective compositions and methods for cancer treatment, optional in combination with another existing cancer treatment. The present invention provides strategies to counteract tumor-induced immune tolerance and enhance the antitumor efficacy of chemotherapy by activating and leveraging T cell-mediated adaptive antitumor immunity against resistant or disseminated cancer cells.

Claims

1. An isolated molecule comprising a targeting moiety fused with an immunomodulatory moiety, wherein:

(a) the targeting moiety specifically binds Cytotoxic T-lymphocyte associated antigen-4 (CTLA-4); and

(b) the immunomodulatory moiety comprises an amino acid sequence of the extracellular domain of Transforming growth factor-beta receptor (TGF-β

R).

2. The molecule of claim 1, wherein the targeting moiety comprises an antibody, antibody fragment, scFv, or Fc-containing polypeptide that specifically binds CTLA-4.

3. The molecule of claim 1, wherein the immunomodulatory moiety comprises an amino acid sequence of the extracellular domain of Transforming growth factor-beta receptor selected from the group consisting of SEQ ID NO:

79-91.

4. The molecule of claim 3, wherein the immunomodulatory moiety comprises an amino acid sequence of the extracellular domain of Transforming growth factor-beta receptor II (TGF-β

RII).

5. The molecule of claim 4, wherein the immunomodulatory moiety comprises the amino acid sequence corresponding to SEQ ID NO:

87 or a binding fragment thereof.

6. The molecule of claim 5, wherein the molecule comprises the amino acid sequence corresponding to SEQ ID NO:

5 or a binding fragment thereof.

7. The molecule of claim 6, wherein the molecule comprises the amino acid sequence corresponding to SEQ ID NO:

5 or a binding fragment thereof and the amino acid sequence corresponding to SEQ ID NO;

74 or a binding fragment thereof.

8. A method of treating a neoplastic disease comprising:

administration to a subject in need thereof one or more molecules of claim 1, wherein the neoplastic disease is bladder cancer or melanoma.

9. The method of claim 8, wherein the subject is administered in combination with another anticancer therapy.

10. The method of claim 8, wherein the anticancer therapy comprises a chemotherapeutic molecule, antibody, small molecule kinase inhibitor, hormonal agent, ionizing radiation, ultraviolet radiation, cryoablation, thermal ablation, or radiofrequency ablation.

11. The method of claim 8, wherein the subject is administered in combination with a vaccine.

12. The method of claim 8, wherein the subject is human.

13. The isolated molecule of claim 1, further comprising a linker at the C_H3 region of the Fc.

14. The isolated molecule of claim 13, wherein the linker is (GGGGS)n (SEQ ID NO:

104), and wherein n is 1, 2, 3, 4, 5, 6, 7 or 8.

15. The isolated molecule of claim 13, wherein the linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:

104).