Antibodies against claudin 18.2 useful in cancer diagnosis

Patent metadata

Field	Value
Publication number	US-10053512-B2
Application number	US-201615227565-A
Country	US
Kind code	B2
Filing date	Aug 3, 2016
Priority date	May 9, 2012
Publication date	Aug 21, 2018
Grant date	Aug 21, 2018

Abstract

The invention relates to antibodies directed against an epitope located within the C-terminal portion of CLDN18.2 which are useful, for example, in diagnosing cancer and/or in determining whether cancer cells express CLDN18.2.

First claim

The invention claimed is:

1. An antibody or antigen binding fragment thereof that binds to claudin 18.2 (CLDN18.2) or a peptide having the amino acid sequence TEDEVQSYPSKHDYV (SEQ ID NO: 5) or EVQSYPSKHDYV (SEQ ID NO: 6)
wherein the antibody or antigen-binding fragment comprises
(i) an antibody heavy chain comprising:
a CDR1 having the amino acid sequence set forth in SEQ ID NO: 8,
a CDR2 having the amino acid sequence set forth in SEQ ID NO: 9, and
a CDR3 having the amino acid sequence set forth in SEQ ID NO: 10; and

an antibody light chain comprising:
a CDR1 having the amino acid sequence set forth in SEQ ID NO: 12,
a CDR2 having the amino acid sequence set forth in SEQ ID NO: 13, and
a CDR3 having the amino acid sequence set forth in SEQ ID NO: 14; or

(ii) an antibody heavy chain comprising:
a CDR1 having the amino acid sequence set forth in SEQ ID NO: 16,
a CDR2 having the amino acid sequence set forth in SEQ ID NO: 17, and
a CDR3 having the amino acid sequence set forth in SEQ ID NO: 18; and

an antibody light chain comprising:
a CDR1 having the amino acid sequence set forth in SEQ ID NO: 20,
a CDR2 having the amino acid sequence set forth in SEQ ID NO: 21, and
a CDR3 having the amino acid sequence set forth in SEQ ID NO: 22.

2. The antibody or antigen-binding fragment thereof of claim 1 , wherein said CLDN18.2 is cell surface membrane-bound CLDN18.2.

3. The antibody or antigen-binding fragment thereof of claim 1 , wherein said CLDN18.2 is present on cancer cells.

4. The antibody or antigen-binding fragment thereof of claim 3 , wherein said cancer cells are CLDN18.2 expressing cancer cells.

5. The antibody or antigen-binding fragment thereof of claim 3 , wherein said cancer cells are selected from the group consisting of gastric, esophageal, pancreatic, lung, ovarian, colon, hepatic, head-neck, and gallbladder cancer cells.

6. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof does not bind to non-cancerous cells except stomach epithelial cells.

7. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof does not bind to non-cancerous lung cells.

8. The antibody of claim 1 , wherein the antibody is a chimeric, human or humanized antibody.

9. The antibody of claim 1 , wherein the antibody is a monoclonal antibody.

10. A conjugate comprising an antibody or antigen-binding fragment of claim 1 coupled to at least one detectable label.

11. A hybridoma capable of producing the antibody of claim 1 .

12. The antibody or antigen-binding fragment of claim 1 , wherein said CLDN18.2 comprises the amino acid sequence according to SEQ ID NO: 2 of the sequence listing or a variant of said amino acid sequence.

13. The antibody or antigen-binding fragment of claim 1 , wherein the antibody or antigen-binding fragment comprises
an antibody heavy chain comprising:
a CDR1 having the amino acid sequence set forth in SEQ ID NO: 16,
a CDR2 having the amino acid sequence set forth in SEQ ID NO: 17, and
a CDR3 having the amino acid sequence set forth in SEQ ID NO: 18; and

an antibody light chain comprising:
a CDR1 having the amino acid sequence set forth in SEQ ID NO: 20,
a CDR2 having the amino acid sequence set forth in SEQ ID NO: 21, and
a CDR3 having the amino acid sequence set forth in SEQ ID NO: 22.

14. The antibody or antigen-binding fragment of claim 1 , wherein the antibody or antigen-binding fragment comprises
an antibody heavy chain comprising:
a CDR1 having the amino acid sequence set forth in SEQ ID NO: 8,
a CDR2 having the amino acid sequence set forth in SEQ ID NO: 9, and
a CDR3 having the amino acid sequence set forth in SEQ ID NO: 10; and

an antibody light chain comprising:
a CDR1 having the amino acid sequence set forth in SEQ ID NO: 12,
a CDR2 having the amino acid sequence set forth in SEQ ID NO: 13, and
a CDR3 having the amino acid sequence set forth in SEQ ID NO: 14.

15. The antibody or antigen-binding fragment of claim 13 , wherein the antibody or antigen-binding fragment comprises an antibody heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 15, and an antibody light chain comprising the amino acid sequence set forth in SEQ ID NO: 19.

16. The antibody or antigen-binding fragment of claim 14 , wherein the antibody or antigen-binding fragment comprises an antibody heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 7, and an antibody light chain comprising the amino acid sequence set forth in SEQ ID NO: 11.

17. A diagnostic test kit which comprises the antibody or antigen-binding fragment of claim 1 .

18. A method for detecting CLDN18.2 or determining the quantity of CLDN18.2 in a sample comprising the steps of:
(i) contacting a sample with the antibody or antigen-binding fragment of claim 1 and
(ii) detecting the formation of a complex or determining the quantity of a complex between the antibody or the antigen-binding fragment and CLDN18.2.

19. A method for determining whether cells express CLDN18.2 comprising the steps of:
(i) contacting a cellular sample with the antibody or antigen-binding fragment of claim 1 and
(ii) detecting the formation of a complex between the antibody or the antigen-binding fragment and CLDN18.2 expressed by cells in said sample.

20. A method for diagnosis, detection or monitoring of cancer comprising the steps of:
contacting a biological sample with the antibody or antigen-binding fragment of claim 1 and
(ii) detecting the formation of a complex and/or determining the quantity of a complex between the antibody or the antigen-binding fragment and CLDN18.2.

21. A method for determining whether a cancer is treatable by a cancer therapy targeting CLDN18.2 comprising the steps of:
(i) contacting a sample comprising cancer cells with the antibody or antigen-binding fragment of claim 1 and
(ii) detecting the formation of a complex between the antibody or the antigen-binding fragment and CLDN18.2.

22. The method of claim 18 , wherein the sample comprises a paraffin embedded tissue section fixed with paraformaldehyde.

23. The method of claim 18 , wherein, in step (i), the antibody or antigen binding fragment is at a concentration of about 0.1 to about 0.5 μg/ml.

24. The method of claim 23 , wherein the concentration is about 0.2 μg/ml.

25. The method of claim 18 , wherein step (i) comprises incubating the sample with the antibody or antigen-binding fragment overnight at 4° C.

26. The method of claim 18 , wherein the antibody or antigen binding fragment is labeled with a detectable label.

27. The method of claim 26 , wherein the detectable label is an enzyme or a fluorophore.

28. The method of claim 18 , wherein the method further comprises contacting the antibody or antigen-binding fragment with a secondary antibody, wherein the secondary antibody is labeled with a detectable label.

29. The method of claim 28 , wherein the secondary antibody is a horseradish-peroxidase (HRP)-labeled secondary antibody.

30. The method of claim 29 , wherein step (ii) comprises exposing the HRP-labeled secondary antibody to a chemiluminescent substrate.

31. The method of claim 18 , wherein the antibody or antigen-binding fragment comprises
an antib

Assignees

• Ganymed Pharmaceuticals Ag
• Tron—Translationale Onkologie An Der Univ Der Johannes Gutenberg Univ Mainz Ggmbh
• Tron—Translationale Onkologie An Der Univ Der Johannes Gutenberg—Universitat Mainz Gem

Inventors

• Sahin Ugur
• Tureci Ozlem
• Mitnacht-Kraus Rita
• Woll Stefan

Classifications

• A61P37/00 Primary

  Drugs for immunological or allergic disorders · CPC title

• G01N33/5759

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

• C07K2317/34

  Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues · CPC title

• C07K16/303

  Liver or Pancreas · CPC title

• C07K16/3046

  Stomach, Intestines · CPC title