Constructs specifically recognizing glypican 3 and uses thereof

Patent metadata

Field	Value
Publication number	US-11447564-B2
Application number	US-201816608377-A
Country	US
Kind code	B2
Filing date	Apr 24, 2018
Priority date	Apr 26, 2017
Publication date	Sep 20, 2022
Grant date	Sep 20, 2022

Abstract

The present application provides constructs comprising an antibody moiety specifically recognizing Glypican 3 (GPC3), such as a cell surface-bound GPC3. Also provided are methods of making and using these constructs.

First claim

What is claimed is:

1. An isolated anti-Glypican-3 (anti-GPC3) construct comprising an antibody moiety specifically recognizing GPC3 (anti-GPC3 antibody moiety), wherein the anti-GPC3 antibody moiety comprises:
(1) a heavy chain complementarity determining region (HC-CDR) 1 comprising the amino acid sequence of SEQ ID NO: 3, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 54, an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 105, a light chain complementarity determining region (LC-CDR) 1 comprising the amino acid sequence of SEQ ID NO: 156, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 207, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 258;
(2) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 72, an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 123, an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 174, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 225, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 276;
(3) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 75, an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 126, an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 177, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 228, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 279;
(4) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 52, an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 103, an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 154, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 205, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 256;
(5) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 62, an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 113, an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 164, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 215, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 266; or
(6) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 63, an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 114, an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 165, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 216, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 267.

2. The isolated anti-GPC3 construct of claim 1 , wherein the anti-GPC3 antibody moiety comprises:
(1) i) a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO: 309, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 309; and ii) a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO: 360, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 360;
(2) i) a VH comprising the amino acid sequence of SEQ ID NO: 327, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 327; and ii) a VL comprising the amino acid sequence of SEQ ID NO: 378, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 378;
(3) i) a VH comprising the amino acid sequence of SEQ ID NO: 330, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 330 and ii) a VL comprising the amino acid sequence of SEQ ID NO: 381, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 381;
(4) i) a VH comprising the amino acid sequence of SEQ ID NO: 307, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 307; and ii) a VL comprising the amino acid sequence of SEQ ID NO: 358, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 358; or
(5) i) a VH comprising the amino acid sequence of SEQ ID NO: 317, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 317; and ii) a VL comprising the amino acid sequence of SEQ ID NO: 368, or a variant thereof having at least about 95% sequence identity to SEQ ID NO: 368.

3. The isolated anti-GPC3 construct of claim 1 , wherein the anti-GPC3 antibody moiety is fused to an Fc fragment optionally via a linker.

4. The isolated anti-GPC3 construct of claim 1 , wherein the isolated anti-GPC3 construct is a tandem scFv comprising two scFvs linked by a peptide linker.

5. The isolated anti-GPC3 construct of claim 1 , wherein the isolated anti-GPC3 construct is a chimeric antigen receptor (CAR) comprising:
(a) an extracellular domain comprising the anti-GPC3 antibody moiety;
(b) a transmembrane domain; and
(c) an intracellular signaling domain.

6. The isolated anti-GPC3 construct of claim 1 , wherein the isolated anti-GPC3 construct is a chimeric antibody-T cell receptor (TCR) construct (caTCR) comprising:
(a) an extracellular domain comprising the anti-GPC3 antibody moiety; and
(b) a TCR module (TCRM) comprising i) a first TCR domain (TCRD) comprising a first TCR transmembrane domain (TCR-TM) and ii) a second TCRD comprising a second TCR-TM, wherein the TCRM facilitates recruitment of at least one TCR-associated signaling molecule.

7. The isolated anti-GPC3 construct of claim 1 , wherein the isolated anti-GPC3 construct is an immunoconjugate comprising the anti-GPC3 antibody moiety and an effector molecule.

8. The isolated anti-GPC3 construct of claim 7 , wherein the effector molecule is a label.

9. An isolated nucleic acid encoding the polypeptide components of the isolated anti-GPC3 construct of claim 8 .

10. An isolated host cell comprising the isolated anti-GPC3 construct of claim 8 .

11. An effector cell expressing the isolated anti-GPC3 construct of claim 5 .

12. A pharmaceutical composition comprising the isolated anti-GPC3 construct of claim 8 , and a pharmaceutically acceptable carrier.

13. A kit comprising the isolated anti-GPC3 construct of claim 1 .

14. A method of treating an individual having a GPC3-positive cancer, comprising administering to the individual an effective amount of the isolated anti-GPC3 construct of claim 8 , wherein:
i) the isolated anti-GPC3 construct is expressed on the cell surface of an immune effector cell;
ii) the isolated anti-GPC3 construct further comprises a second antigen-binding moiety specifically recognizing a cell surface antigen on an immune effector cell; or
iii) the isolated anti-GPC3 construct further comprises an effector molecule, wherein the effector molecule is a therapeutic agent selected from the group consisting of a drug, a toxin, a radioisotope, a protein, a peptide, and a nucleic acid.

15. A method of diagnosing an individual having a GPC3-positive disease, comprising:
a) administering an effective amount of the isolated anti-GPC3 construct of claim 8 to the individual; and
b) determining the level of the label in the individual, wherein a level of the label above a threshold level indicates that the individual has the GPC3-positive disease.

16. A method of diagnosing an individual having a GPC3-positive disease, comprising:
a) contacting a sample derived from the individual with the isolated anti-GPC3 construct of claim 8 ; and
b) determining the number of cells bound with the isolated anti-GPC3 construct in the sample, wherein a value for the number of cells bound with the isolated anti-GPC3 construct above a threshold lev

Assignees

• Eureka Therapeutics Inc

Inventors

• Zhang Pengbo
• Xu Yiyang
• Morales Javier
• Nakano Yoko
• Liu Hong
• Xiang Jingyi
• Acker Timothy

Classifications

• G01N33/57525

  of the liver or pancreas · CPC title

• G01N33/575

  for cancer · CPC title

• G01N33/5759 Primary

  involving compounds localised on the membrane of tumour or cancer cells · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K14/70521

  CD28, CD152 · CPC title