Compositions and methods for rendering tumor cells susceptible to CD8+ T cell-mediated killing

Patent metadata

Field	Value
Publication number	US-10183985-B2
Application number	US-201715403593-A
Country	US
Kind code	B2
Filing date	Jan 11, 2017
Priority date	Sep 14, 2012
Publication date	Jan 22, 2019
Grant date	Jan 22, 2019

Abstract

The present invention provides an immunoconjugate having the formula:

T-c-E n -c-Fc n or T-c-Fc n -c-E n ;

wherein, T is a single chain variable portion fragment of a monoclonal antibody (scFv) directed to a target protein, polypeptide, or fragment thereof, which is highly expressed on cancer cells; E is two or more foreign immunogenic CD8 + T cell antigenic epitopes; c is a peptide or polypeptide fragment thereof, capable of being cleaved by a specific protease; and Fc is two or more Fc portions of an IgG antibody. Nucleic acid sequences encoding the same and vectors containing said nucleic acid sequences are also provided. Methods of making the immunoconjugate, along with methods of making target cells susceptible to CTL mediated cell killing, and methods for treatment of cancers are also provided.

First claim

The invention claimed is:

1. A method for making a tumor cell susceptible to CD8 + T cell killing, comprising contacting one or more tumor cells with an effective amount of an immunoconjugate having the formula:
T-c-E n -c-Fc n ;
wherein
T is a single chain variable portion fragment of a monoclonal antibody (scFv) directed to a target protein, polypeptide, or fragment thereof, which is highly expressed on cancer cells, and selected from the group consisting of scFv directed to mesothelin, epidermal growth factor receptor (EGFR), NKG2D, or Her2/neu protein;
E n is two or more foreign immunogenic CD8 + T cell antigenic epitopes derived from ovalbumin, Epstein-Barr virus, cytomegalovirus, human papilloma virus, and influenza wherein n is 1 to 10;
c is a peptide or polypeptide fragment thereof, capable of being cleaved by by furin, MMP1 or MMP9; and
Fc n is two or more Fc portions of an IgG antibody wherein n is 1 to 10.

2. A method for making a tumor cell susceptible to CD8 + T cell killing, comprising contacting one or more tumor cells with an effective amount of an immunoconjugate having the formula:
T-c-Fc n -c-E n ;
wherein
T is a single chain variable portion fragment of a monoclonal antibody (scFv) directed to a target protein, polypeptide, or fragment thereof, which is highly expressed on cancer cells, and selected from the group consisting of scFv directed to mesothelin, epidermal growth factor receptor (EGFR), NKG2D, or Her2/neu protein;
E n is two or more foreign immunogenic CD8 + T cell antigenic epitopes derived from ovalbumin, Epstein-Barr virus, cytomegalovirus, human papilloma virus, and influenza wherein n is 1 to 10;
c is a peptide or polypeptide fragment thereof, capable of being cleaved by by furin, MMP1 or MMP9; and
Fc n is two or more Fc portions of an IgG antibody wherein n is 1 to 10.

3. The method of claim 1 , wherein c is a furin cleavable peptide having the amino acid sequence RVKR (SEQ ID NO: 2).

4. The method of claim 2 , wherein c is a furin cleavable peptide having the amino acid sequence RVKR (SEQ ID NO: 2).

5. The method of claim 1 , wherein E is an ovalbumin epitope having the amino acid sequence SIINFEKL (SEQ ID NO: 1).

6. The method of claim 2 , wherein E is an ovalbumin epitope having the amino acid sequence SIINFEKL (SEQ ID NO: 1).

7. The method of claim 1 , wherein T is a scFv directed to mesothelin.

8. The method of claim 2 , wherein T is a scFv directed to mesothelin.

9. The method of claim 1 , wherein the method further comprises, determining whether the CD8 + T cell antigenic epitopes is specific for an antigen presented on the tumor cell and then contacting one or more tumor cells with the immunoconjugate of claim 1 having said antigenic epitope.

10. The method of claim 2 , wherein the method further comprises, determining whether the CD8 + T cell antigenic epitopes is specific for an antigen presented on the tumor cell and then contacting one or more tumor cells with the immunoconjugate of claim 2 having said antigenic epitope.

11. The method of claim 1 , wherein the tumor cell is a cancer cell.

12. The method of claim 2 , wherein the tumor cell is a cancer cell.

13. The method of claim 11 , wherein the tumor cell is in a subject.

14. The method of claim 12 , wherein the tumor cell is in a subject.

Assignees

• Univ Johns Hopkins

Inventors

• Hung Chien-Fu
• Wu T C

Classifications

• A61K2039/585

  wherein the target is cancer · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• A61K2039/505

  comprising antibodies · CPC title

• C07K2319/50

  containing protease site · CPC title

• A61K2039/627

  characterised by the linker · CPC title