Glypican-3 antibody and uses thereof

Patent metadata

Field	Value
Publication number	US-10093746-B2
Application number	US-201515508317-A
Country	US
Kind code	B2
Filing date	Sep 3, 2015
Priority date	Sep 4, 2014
Publication date	Oct 9, 2018
Grant date	Oct 9, 2018

Abstract

The present invention relates to compositions and methods of isolated polynucleotides that encode or polypeptides comprising glypican-3 (GPC3). The invention also includes a chimeric antigen receptor (CAR) wherein the CAR is able to target GPC3. The invention further includes methods of treating a subject or diagnosing and treating diseases, disorders or conditions associated with dysregulated glypican-3.

First claim

What is claimed is:

1. An isolated polynucleotide encoding an anti-glypican-3 (GPC3) antibody or antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises SEQ ID NO: 1 and the light chain variable region comprises SEQ ID NO: 3.

2. An isolated anti-GPC3 antibody or antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises SEQ ID NO: 2 and the light chain variable region comprises SEQ ID NO: 4.

3. The antigen-binding fragment of claim 2 , wherein the antigen-binding fragment is selected from the group consisting of a Fab fragment, a F(ab′) 2 fragment, a Fv fragment, and a single chain Fv (scFv).

4. A method of imaging or visualizing a sample taken from a normal or malignant tissue, the method comprising contacting the sample with a labeled anti-GPC3 antibody or antigen-binding fragment thereof, wherein the anti-GPC3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises SEQ ID NO: 2 and the light chain variable region comprises SEQ ID NO: 4.

5. The method of claim 4 , wherein the tissue comprises liver tissue.

6. The method of claim 4 , wherein the antigen-binding fragment is a Fab or a scFv.

7. A method of inhibiting growth of a GPC3-expressing tumor cell, the method comprising contacting the tumor cell with an anti-GPC3 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein an amino acid sequence of the heavy chain variable region comprises SEQ ID NO: 2 and the amino acid sequence of the light chain variable region comprises SEQ ID NO: 4.

8. A pharmaceutical composition comprising an effective amount of the isolated polynucleotide of claim 1 .

9. A pharmaceutical composition comprising an effective amount of the isolated anti-GPC3 antibody or antigen-binding fragment thereof of claim 2 .

10. An isolated nucleic acid sequence encoding a chimeric antigen receptor (CAR) comprising a glypican-3 (GPC3) binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein the isolated nucleic acid sequence that encodes the GPC3 binding domain comprises SEQ ID NO: 1 and SEQ ID NO: 3.

11. The isolated nucleic acid sequence of claim 10 , wherein the encoded GPC3 binding domain comprises SEQ ID NO: 2 and SEQ ID NO: 4.

12. The isolated nucleic acid sequence of claim 10 , wherein the co-stimulatory signaling domain comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, and any combination thereof.

13. The isolated nucleic acid sequence of claim 11 , wherein the encoded GPC3 binding domain is selected from the group consisting of a Fab fragment, a F(ab′) 2 fragment, a Fv fragment, and a single chain Fv (scFv).

14. An isolated chimeric antigen receptor (CAR) comprising a GPC3 binding domain and a CD3 zeta signaling domain, wherein the GPC3 binding domain comprises SEQ ID NO: 2 and SEQ ID NO: 4.

15. The isolated CAR of claim 14 , further comprising the sequence of a costimulatory signaling domain.

16. The isolated CAR of claim 15 , wherein the co-stimulatory signaling domain is selected from the group consisting of CD27, CD28, 4-1BB 0X40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, and any combination thereof.

17. The isolated CAR of claim 14 , wherein the GPC3 binding domain is a humanized antibody or antigen binding fragment thereof.

18. The isolated CAR of claim 14 , wherein the GPC3 binding domain is selected from the group consisting of a Fab fragment, a F(ab′) 2 fragment, a Fv fragment, and a single chain Fv (scFv).

19. A T cell comprising the isolated nucleic acid sequence of claim 10 or the CAR of claim 14 .

20. A method of providing an anti-tumor immunity in a subject with a tumor that expresses glypican-3 (GPC3), the method comprising administering to the subject an effective amount of a genetically modified T cell comprising an isolated nucleic acid sequence encoding a chimeric antigen receptor (CAR) that comprises a glypican-3 (GPC3) binding domain, a 4-1BB co-stimulatory signaling domain, and a CD3 zeta signaling domain, wherein the GPC3 binding domain comprises SEQ ID NO: 2 and SEQ ID NO: 4.

21. The method of claim 20 , wherein the T cell is an autologous T cell.

22. The method of claim 20 , wherein the subject is a human.

23. A method of treating a glypican-3 (GPC3)-expressing liver cancer in a subject, the method comprising administering to the subject an effective amount of a genetically modified T cell comprising an isolated nucleic acid sequence encoding a chimeric antigen receptor (CAR) that comprises a GPC3 binding domain, a 4-1BB co-stimulatory signaling domain, and a CD3 zeta signaling domain, wherein the GPC3 binding domain comprises SEQ ID NO: 2 and SEQ ID NO: 4.

Assignees

• Univ Pennsylvania
• Us Veterans Affairs

Inventors

• Kaplan David

Classifications

• C07K14/7051

  T-cell receptor (TcR)-CD3 complex · CPC title

• C07K16/303 Primary

  Liver or Pancreas · CPC title

• C07K2317/569

  Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody® · CPC title

• A61K2035/124

  the cells being hematopoietic, bone marrow derived or blood cells · CPC title

• C07K2319/03

  containing a transmembrane segment · CPC title