HUMAN MONOCLONAL ANTIBODIES SPECIFIC FOR GLYPICAN-3 AND USE THEREOF

• US 20140044714A1
• Filed: 04/19/2012
• Published: 02/13/2014
• Est. Priority Date: 04/19/2011
• Status: Active Grant

• Current Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Original Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Inventors: Dimitrov, Dimiter S., Ho, Mitchell, Feng, Mingqian, Gao, Wei, Kim, Heungnam
• Granted Patent: US 9,206,257 B2
• Time in Patent Office: Days
• US Class Current: 424/134.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• CPC Class Codes: A61K 47/6813 the drug being a peptidic cytokine, e.g. an interleukin or interferon
• CPC Class Codes: A61K 47/6829 Bacterial toxins, e.g. diphteria toxins or Pseudomonas exotoxin A
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6851 the antibody targeting a determinant of a tumour cell
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/21 from Pseudomonadaceae (F)
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/303 Liver or Pancreas
• CPC Class Codes: C07K 16/3076 against structure-related tumour-associated moieties
• CPC Class Codes: C07K 16/3092 against tumour-associated mucins
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/569 Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
• 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 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/55 : containing a fusion with a toxin, e.g. diphteria toxin
• CPC Class Codes: G01N 2333/705 : Assays involving receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 2400/40 : Glycosaminoglycans, i.e. GAG or mucopolysaccharides, e.g. chondroitin sulfate, dermatan sulfate, hyaluronic acid, heparin, heparan sulfate, and related sulfated polysaccharides
• CPC Class Codes: G01N 33/57438 : of liver, pancreas or kidney
• CPC Class Codes: G01N 33/57492 : involving compounds localized on the membrane of tumor or cancer cells
• CPC Class Codes: G01N 33/6893 : related to diseases not provided for elsewhere

Abstract

Described herein is the identification of human monoclonal antibodies that bind GPC3 or heparan sulfate (HS) chains on GPC3 with high affinity. The antibodies described herein are capable of inhibiting HCC cell growth and migration. Provided are human monoclonal antibodies specific for GPC3 or HS chains on GPC3, including immunoglobulin molecules, such as IgG antibodies, as well as antibody fragments, such as single-domain VH antibodies or single chain variable fragments (scFv). Further provided are compositions including the antibodies that bind GPC3 or HS chains on GPC3, nucleic acid molecules encoding these antibodies, expression vectors comprising the nucleic acids, and isolated host cells that express the nucleic acids. Methods of treating cancer and/or inhibiting tumor growth or metastasis are also provided. Further provided are methods of detecting cancer in a subject and confirming a diagnosis of cancer in a subject.

Claims

1. An isolated human monoclonal antibody that binds glypican-3 (GPC3), wherein the heavy chain of the antibody comprises amino acid residues 26-33 of SEQ ID NO:

2, amino acid residues 51-57 of SEQ ID NO;

2, and amino acid residues 96-105 of SEQ ID NO;

2.

2. The human monoclonal antibody of claim 1, wherein the heavy chain of the antibody comprises the amino acid sequence of SEQ ID NO:

2.

8. The human monoclonal antibody of claim 1, wherein the antibody is a VH single-domain antibody, a Fab fragment, a Fab′

fragment, a F(ab)′

₂ fragment, a single chain variable fragment (scFv), or a disulfide stabilized variable fragment (dsFv).

9. The human monoclonal antibody of claim 8, wherein the antibody is a VH single-domain antibody.

10. The human monoclonal antibody of claim 8, wherein the antibody is a scFv.

11. The human monoclonal antibody of claim 1, wherein the antibody is an IgG.

12. The isolated human monoclonal antibody of claim 1, wherein the antibody is labeled.

13. The isolated human monoclonal antibody of claim 12, wherein the label is a fluorescence, enzymatic, or radioactive label.

14. A composition comprising a therapeutically effective amount of the antibody of claim 1 in a pharmaceutically acceptable carrier.

15. An isolated immunoconjugate comprising the human monoclonal antibody of claim 1 and an effector molecule.

16. The isolated immunoconjugate of claim 15, wherein the effector molecule is a toxin.

17. The isolated immunoconjugate of claim 16, wherein the toxin is Pseudomonas exotoxin or a variant thereof.

18. The isolated immunoconjugate of claim 17, wherein the toxin is PE38 comprising the amino acid sequence of SEQ ID NO:

27.

20. The isolated immunoconjugate of claim 18, comprising the amino acid sequence of SEQ ID NO:

29.

22. A composition comprising a therapeutically effective amount of the isolated immunoconjugate of claim 15 in a pharmaceutically acceptable carrier.

23. A method of treating a subject with cancer, comprising selecting a subject with a cancer that expresses GPC3 and administering to the subject a therapeutically effective amount of the composition of claim 14, thereby treating the cancer in the subject.

24. A method of inhibiting tumor growth or metastasis, comprising selecting a subject with a cancer that expresses GPC3 and administering to the subject a therapeutically effective amount of the composition of claim 14, thereby inhibiting tumor growth or metastasis.

25. A method of determining if a subject has cancer, comprising:

contacting a sample from the subject with the human monoclonal antibody of claim 1; and

detecting binding of the antibody to the sample,

wherein an increase in binding of the antibody to the sample as compared to binding of the antibody to a control sample identifies the subject as having cancer.

26. A method of confirming a diagnosis of cancer in a subject, comprising:

contacting a sample from a subject diagnosed with cancer with the human monoclonal antibody of claim 1; and

detecting binding of the antibody to the sample,

wherein an increase in binding of the antibody to the sample as compared to binding of the antibody to a control sample confirms the diagnosis of cancer in the subject.

29. The method of claim 23, wherein the cancer is hepatocellular carcinoma (HCC), melanoma, lung cancer or ovarian cancer.

30. The method of claim 29, wherein the cancer is HCC.

33. An isolated nucleic acid molecule encoding the human monoclonal antibody of claim 1.

34. The isolated nucleic acid molecule of claim 33, wherein the nucleotide sequence encoding the heavy chain of the human monoclonal antibody comprises SEQ ID NO:

1.

37. The isolated nucleic acid molecule of claim 33, operably linked to a promoter.

38. An expression vector comprising the isolated nucleic acid molecule of claim 33.

39. An isolated host cell transformed with the nucleic acid molecule or expression vector of claim 33.

3. An isolated human monoclonal antibody that binds heparan sulfate (HS) on GPC3, wherein:

(i) the heavy chain of the antibody comprises amino acid residues 26-33 of SEQ ID NO;

14, amino acid residues 51-58 of SEQ ID NO;

14, and amino acid residues 97-105 of SEQ ID NO;

14; and

(ii) the light chain of the antibody comprises amino acid residues 27-32 of SEQ ID NO;

16 or SEQ ID NO;

31, amino acid residues 50-52 of SEQ ID NO;

16 or SEQ ID NO;

31, and amino acid residues 89-97 of SEQ ID NO;

16 or SEQ ID NO;

31.

6. The human monoclonal antibody of claim 3, wherein the heavy chain of the antibody comprises the amino acid sequence of SEQ ID NO:

14, and the light chain of the antibody comprises the amino acid sequence of SEQ ID NO;

16 or SEQ ID NO;

31.

35. An isolated nucleic acid molecule encoding the human monoclonal antibody of claim 3.

36. The isolated nucleic acid molecule of claim 35, wherein the nucleotide sequence encoding the heavy chain of the human monoclonal antibody comprises SEQ ID NO:

13, and the nucleotide sequence encoding the light chain of the human monoclonal antibody comprises SEQ ID NO;

15 or SEQ ID NO;

30.

4-5. -5. (canceled)

7. (canceled)

19. (canceled)

21. (canceled)

27-28. -28. (canceled)

31-32. -32. (canceled)

40-52. -52. (canceled)