Anti-Pancreatic Cancer Antibodies

• US 20090304580A1
• Filed: 08/07/2009
• Published: 12/10/2009
• Est. Priority Date: 06/14/2002
• Status: Active Grant

• Current Assignee: Immunomedics, Inc.
• Original Assignee: Immunomedics, Inc.
• Inventors: Chang, Chien-Hsing, Goldenberg, David M., Hansen, Hans J., Gold, David V.
• Granted Patent: US 8,491,896 B2
• Time in Patent Office: Days
• US Class Current: 424/1.490
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/507 Comprising a combination of two or more separate antibodies
• CPC Class Codes: A61K 47/6897 Pre-targeting systems with two or three steps using antibody conjugates; Ligand-antiligand therapies
• CPC Class Codes: A61K 51/0406 Amines, polyamines, e.g. spermine, spermidine, amino acids, (bis)guanidines
• CPC Class Codes: A61K 51/0491 Sugars, nucleosides, nucleotides, oligonucleotides, nucleic acids, e.g. DNA, RNA, nucleic acid aptamers
• CPC Class Codes: A61K 51/08 Peptides, e.g. proteins , carriers being peptides, polyamino acids, proteins
• CPC Class Codes: A61K 51/088 conjugates with carriers being peptides, polyamino acids or proteins antibodies A61K51/10
• CPC Class Codes: A61K 51/109 immunoglobulins having two or more different antigen-binding sites or multifunctional antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 16/303 Liver or Pancreas
• CPC Class Codes: C07K 16/3092 against tumour-associated mucins
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]

Abstract

Described herein are compositions and methods of use of anti-pancreatic cancer antibodies or fragments thereof, such as murine, chimeric, humanized or human PAM4 antibodies. The subject antibodies show a number of novel and useful therapeutic characteristics, such as binding with high specificity to pancreatic and other cancers, but not to normal or benign pancreatic tissues and binding to a high percentage of early stage pancreatic cancers. In preferred embodiments, the antibodies bind to pancreatic cancer mucins. The antibodies and fragments are of use for the detection, diagnosis and/or treatment of cancer, such as pancreatic cancer. The antibodies, such as PAM4 antibodies, bind to a PAM4 antigen that shows unique cell and tissue distributions compared with other known antibodies such as CA19.9, DUPAN2, SPAN1, Nd2, B72.3, and Le^a and Le(y) antibodies that bind to the Lewis antigens.

Claims

1. An antibody construct comprising at least one anti-pancreatic cancer antibody or antigen-binding fragment thereof that binds to pancreatic cancer cells and does not bind to normal pancreatic tissue or pancreatitis or benign pancreatic tissue.

2. The antibody construct of claim 1, wherein the anti-pancreatic cancer antibody binds to PanIN-1A, PanIN-1B, PanIN-2, invasive pancreatic adenocarcinoma, pancreatic carcinoma, mucinous cyst neoplasms (MCN), intrapancreatic mucinous neoplasms (IPMN) and intraductal papillary mucinous neoplasia.

3. The antibody construct of claim 1, wherein the construct is a trimeric DNL (dock and lock) construct comprising two copies of a PAM4 antibody fragment moiety attached to one copy of a 679 antibody fragment moiety.

4. The antibody construct of claim 3, wherein each PAM4 antibody fragment moiety comprises a PAM4 Fab fragment attached to a DDD2 (SEQ ID NO:

34) sequence and the 679 antibody fragment moiety comprises a 679 Fab fragment attached to an AD2 (SEQ ID NO;

36) sequence and a dimer of the DDD2 sequence binds to the AD2 sequence to form the trimeric construct.

5. The antibody construct of claim 4, wherein the PAM4 antibody fragment is an hPAM4 Fab fragment and the 679 antibody fragment is an h679 Fab fragment.

6. The antibody construct of claim 3, wherein the PAM4 antibody fragment moiety comprises the light chain variable region CDR sequences CDR1 (SASSSVSSSYLY, (SEQ ID NO:

1);

CDR2 (STSNLAS, SEQ ID NO;

2); and

CDR3 (HQWNRYPYT, SEQ ID NO;

3); and

the heavy chain variable region CDR sequences CDR1 (SYVLH, SEQ ID NO;

4);

CDR2 (YINPYNDGTQYNEKFKG, SEQ ID NO;

5) and CDR3 (GFGGSYGFAY, SEQ ID NO;

6).

7. The antibody construct of claim 3, wherein the PAM4 antibody fragment moiety comprises a murine, chimeric, humanized or human antibody PAM4 fragment.

8. The antibody construct of claim 1, wherein the anti-pancreatic cancer antibody or fragment thereof binds to the PAM4 antigen.

9. The antibody construct of claim 1, wherein the anti-pancreatic cancer antibody or fragment thereof competes for binding to human pancreatic cancer mucin with a chimeric PAM4 (cPAM4) antibody, wherein the cPAM4 antibody comprises the light chain variable region CDR sequences CDR1 (SASSSVSSSYLY, (SEQ ID NO:

1);

CDR2 (STSNLAS, SEQ ID NO;

2); and

CDR3 (HQWNRYPYT, SEQ ID NO;

3); and

the heavy chain variable region CDR sequences CDR1 (SYVLH, SEQ ID NO;

4);

CDR2 (YINPYNDGTQYNEKFKG, SEQ ID NO;

5) and CDR3 (GFGGSYGFAY, SEQ ID NO;

6).

10. The antibody construct of claim 1, wherein the anti-pancreatic cancer antibody or fragment thereof binds to a linear peptide comprising the amino acid sequence WTWNITKAYPLP (SEQ ID NO:

29) or to a cyclic peptide comprising the amino acid sequence ACPEWWGTTC (SEQ ID NO;

30).

42. A method of diagnosing pancreatic cancer comprising:

a) obtaining an antibody according to claim 1, wherein the antibody is either labeled with at least one diagnostic agent or the antibody is part of a bispecific antibody that binds to a targetable construct labeled with at least one diagnostic agent;

b) exposing the antibody to a sample suspected of containing pancreatic cancer cells; and

c) detecting the presence of antibody bound to pancreatic cancer cells in the sample, wherein binding of antibody to the sample indicates the presence of pancreatic cancer.

43. The method of claim 42, wherein binding of antibody to the sample distinguishes the presence of pancreatic cancer from normal or benign pancreatic tissue in the sample.

44. The method of claim 42, wherein binding of antibody to the sample distinguishes the presence of pancreatic cancer from pancreatitis in the sample.

45. The method of claim 42, wherein binding of antibody to the sample is diagnostic of the presence of early stage pancreatic cancer in the sample.

46. The method of claim 42, wherein the antibody is exposed to the sample in vivo or in vitro.

47. The method of claim 42, wherein the sample is a serum sample and the method further comprises performing an organic phase extraction on the serum sample before the sample is exposed to the antibody.

48. The method of claim 47, wherein the organic phase is butanol.

11. A method of treating pancreatic cancer, comprising administering to an individual with pancreatic cancer an antibody construct comprising at least one anti-pancreatic cancer antibody or antigen-binding fragment thereof that binds to pancreatic cancer cells and does not bind to normal pancreatic tissue or pancreatitis or benign pancreatic tissue.

12. The method of claim 11, wherein the anti-pancreatic cancer antibody or antigen-binding fragment thereof binds to 80% or more of human invasive pancreatic adenocarcinoma, intraductal papillary mucinous neoplasia, PanIN-1A, PanIN-1B and PanIN-2.

13. The method of claim 11, wherein the anti-pancreatic cancer antibody or antigen-binding fragment thereof binds to PanIN-1A, PanIN-1B and PanIN-2 but not to pancreatitis.

14. The method of claim 11, wherein the anti-pancreatic cancer antibody or antigen-binding fragment thereof is a naked antibody or fragment thereof.

15. The method of claim 14, further comprising administering at least one therapeutic agent to the individual.

16. The method of claim 11, wherein the anti-pancreatic cancer antibody or antigen-binding fragment thereof is conjugated to at least one therapeutic agent.

17. The method of claim 16, wherein the therapeutic agent is selected from the group consisting of a radionuclide, an immunomodulator, a hormone, a hormone antagonist, an enzyme, an anti-sense oligonucleotide, an siRNA, an enzyme inhibitor, a photoactive therapeutic agent, a cytotoxic agent, a drug, a toxin, an angiogenesis inhibitor and a pro-apoptotic agent.

18. The method of claim 17, wherein the drug is selected from the group consisting of nitrogen mustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, gemcitabine, triazenes, folic acid analogs, anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzyme inhibitors, epipodophyllotoxins, platinum coordination complexes, vinca alkaloids, substituted ureas, methyl hydrazine derivatives, adrenocortical suppressants, hormone antagonists, endostatin, taxols, camptothecins, SN-38, doxorubicins and their analogs, antimetabolites, alkylating agents, antimitotics, anti-angiogenic agents, tyrosine kinase inhibitors, mTOR inhibitors, heat shock protein (HSP90) inhibitors, proteosome inhibitors, HDAC inhibitors, pro-apoptotic agents, methotrexate and CPT-11.

19. The method of claim 17, wherein the toxin is selected from the group consisting of ricin, abrin, alpha toxin, saporin, ribonuclease (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin, Pseudomonas exotoxin, and Pseudomonas endotoxin.

20. The method of claim 17, wherein the immunomodulator is selected from the group consisting of a cytokine, a stem cell growth factor, a lymphotoxin, a hematopoietic factor, a colony stimulating factor (CSF), an interleukin (IL), an interferon (IFN), a stem cell growth factor, erythropoietin, thrombopoietin, tumor necrosis factor (TNF), granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), interferon-α

, interferon-β

, interferon-γ and

the stem cell growth factor designated “

S1 factor”

.

21. The method of claim 20, wherein the cytokine is selected from the group consisting of human growth hormone, N-methionyl human growth hormone, bovine growth hormone, parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, glycoprotein follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), placenta growth factor (PlGF), hepatic growth factor, prostaglandin, fibroblast growth factor, prolactin, placental lactogen, OB protein, tumor necrosis factor-α

, tumor necrosis factor-β

, mullerian-inhibiting substance, mouse gonadotropin-associated peptide, inhibin, activin, vascular endothelial growth factor, integrin, thrombopoietin (TPO), NGF-β

, platelet-growth factor, TGF-α

, TGF-β

, insulin-like growth factor-I, insulin-like growth factor-II, erythropoietin (EPO), osteoinductive factors, interferon-α

, interferon-β

, interferon-γ

, macrophage-CSF (M-CSF), IL-1, IL-1α

, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21, IL-25, LIF, FLT-3, angiostatin, thrombospondin, endostatin, TNF-α and

LT

22. The method of claim 16, wherein the radionuclide is selected from the group consisting of ¹¹¹In, ¹⁷⁷Lu, ²¹²Bi, ²¹³Bi, ²¹¹At, ⁶²Cu, ⁶⁷Cu, ⁹⁰Y, ¹²⁵I, ¹³¹I, ³²P, ³³P, ⁴⁷Sc, ¹¹¹Ag, ⁶⁷Ga, ¹⁴³Pr, ¹⁵³Sm, ¹⁶¹Tb, ¹⁶⁶Dy, ¹⁶⁶Ho, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁸⁹Re, ²¹²Pb, ²²³Ra, ²²⁵Ac, ⁵⁹Fe, ⁷⁵Se, ⁷⁷As, ⁸⁹Sr, ⁹⁹Mo, ¹⁰⁵Rh, ¹⁰⁹Pd, ¹⁴³Pr, ¹⁴⁹Pm, ¹⁶⁹Er, ¹⁹⁴Ir, ¹⁹⁸Au, ¹⁹⁹Au, ²¹¹Pb, ⁵⁸Co, ^80mBr, ^99mTc, ^103mRh, ¹⁰⁹Pt, ¹¹⁹Sb, ^189mOS, ¹⁹²Ir, ²¹⁹Rn, ²¹⁵Po, ²²¹Fr, ²¹⁷At, ²⁵⁵Fm, ¹¹C, ¹³N, ¹⁵O, ⁷⁵ Br, ²²⁴Ac, ¹²⁶I, ¹³³I, ⁷⁷Br, ^113mIn, ⁹⁵Ru, ⁹⁷Ru, ¹⁰³Ru, ¹⁰⁵Ru, ¹⁰⁷Hg, ²⁰³Hg, ^121mTe, ^122mTe, ^125mTe, ¹⁶⁵Tm, ¹⁶⁷Tm, ¹⁶⁸Tm, ¹⁹⁷Pt, ¹⁴³Pr, ⁵⁷Co, ⁵¹Cr, ⁷⁵Se, ²⁰Tl, ⁷⁶Br and ¹⁶⁹Yb.

23. The method of claim 16, further comprising administering to the individual a second therapeutic agent that is not conjugated to the anti-pancreatic cancer antibody or antigen-binding fragment.

24. The method of claim 23, wherein the anti-pancreatic cancer antibody or antigen-binding fragment thereof is conjugated to a radionuclide and the second therapeutic agent is a radiosensitizing agent that is administered to the individual before the anti-pancreatic cancer antibody or fragment is administered.

25. The method of claim 14, wherein the antibody construct is a bispecific or multispecific antibody comprising at least one PAM4 antibody or antigen-binding fragment thereof and at least one antibody or fragment that binds to a hapten on a targetable construct, and the method further comprises administering a targetable construct attached to at least one therapeutic agent.

26. The method of claim 25, wherein the antibody construct is a trimeric DNL (dock and lock) construct comprising two copies of a PAM4 antibody fragment moiety attached to one copy of a 679 antibody fragment moiety and the targetable construct comprises at least one HSG hapten.

27. The method of claim 11, wherein the antibody construct comprises a second antibody or antigen-binding fragment thereof that binds to a tumor-associated antigen (TAA).

28. The method of claim 27, wherein the second antibody is selected from the group consisting of CA19.9, DUPAN2, SPAN1, Nd2, B72.3, CC49, anti-Le^a antibodies, antibodies to other Lewis antigens such as Le(y), antibodies against carcinoembryonic antigen (CEA), CEACAM6, colon-specific antigen-p (CSAp), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, MUC-16, MUC-17, HLA-DR, CD40, CD74, CD-80, CD138, HER2/neu, EGFR, EGP-1, EGP-2, angiogenesis factors, VEGF, PlGF, insulin-like growth factor (ILGF), carbonic anhydrase IX, tenascin, platelet-derived growth factor, IL-6, oncogene products, bcl-2, Kras, p53, HER2/neu, cMET and antibodies against tumor necrosis substances.

29. A method of detecting or diagnosing pancreatic cancer comprising:

a) administering to an individual an antibody construct comprising at least one anti-pancreatic cancer antibody or antigen-binding fragment thereof that binds to pancreatic cancer cells and does not bind to normal pancreatic tissue or pancreatitis or benign pancreatic tissue; and

b) detecting the antibody construct bound to pancreatic cancer cells.

30. The method of claim 29, wherein the anti-pancreatic cancer antibody binds to PanIN-1A, PanIN-1B, PanIN-2, invasive pancreatic adenocarcinoma, pancreatic carcinoma, mucinous cyst neoplasms (MCN), intrapancreatic mucinous neoplasms (IPMN) and intraductal papillary mucinous neoplasia.

31. The method of claim 29, wherein the anti-pancreatic cancer antibody or antigen-binding fragment thereof binds to 80% or more of human invasive pancreatic adenocarcinoma, intraductal papillary mucinous neoplasia, PanIN-1A, PanIN-1B and PanIN-2.

32. The method of claim 29, wherein the anti-pancreatic cancer antibody or antigen-binding fragment thereof binds to PanIN-1A, PanIN-1B and PanIN-2 but not to pancreatitis.

33. The method of claim 29, wherein the anti-pancreatic cancer antibody or antigen-binding fragment thereof is conjugated to at least one diagnostic agent.

34. The method of claim 33, wherein the diagnostic agent is selected from the group consisting of a radionuclide, a contrast agent, a fluorescent agent, a chemiluminescent agent, a bioluminescent agent, a paramagnetic ion, an enzyme and a photoactive diagnostic agent.

35. The method of claim 34, wherein the diagnostic agent is a radionuclide selected from the group consisting of ¹¹⁰In, ¹¹¹In, ¹⁷⁷Lu, ¹⁸F, ⁵²Fe, ⁶²Cu, ⁶⁴Cu, ⁶⁷Ga, ⁶⁷Ga, ⁶⁸Ga, ⁸⁶Y, ⁹⁰Y, ⁸⁹Zr, ^94mTc, ⁹⁴Tc, ^99mTc, ¹²⁰I, ¹²³I, ¹²⁴I, ¹²⁵I, ¹³¹I, ^154-158Gd, ³²P, ¹¹C, ¹³N, ¹⁵O, ¹⁸⁶Re, ¹⁸⁸Re, ⁵¹Mn, ^52mMn, ⁵⁵Co, ⁷²As, ⁷⁵Br, ⁷⁶Br, ^82mRb, ⁸³Sr, or other gamma-, beta-, or positron-emitters.

36. The method of claim 35, wherein the radionuclide is ¹⁸F and the method further comprising PET imaging.

37. The method of claim 34, wherein the paramagnetic ion is selected from the group consisting of chromium (III), manganese (II), iron (III), iron (II), cobalt (II), nickel (II), copper (II), neodymium (III), samarium (III), ytterbium (III), gadolinium (III), vanadium (II), terbium (III), dysprosium (III), holmium (III) and erbium (III).

38. The method of claim 34, wherein the diagnostic agent is a fluorescent labeling compound selected from the group consisting of fluorescein isothiocyanate, rhodamine, phycoerytherin, phycocyanin, allophycocyanin, o-phthaldehyde and fluorescamine, or a chemiluminescent labeling compound selected from the group consisting of luminol, isoluminol, an aromatic acridinium ester, an imidazole, an acridinium salt and an oxalate ester, or a bioluminescent compound selected from the group consisting of luciferin, luciferase and aequorin.

39. The method of claim 29, wherein the method is used in intraoperative, endoscopic, or intravascular procedure.

40. The method of claim 29, wherein the antibody construct is a trimeric DNL (dock and lock) construct comprising two copies of a PAM4 antibody fragment moiety attached to one copy of a 679 antibody fragment moiety and the method further comprises administering to the individual a targetable construct comprising at least one HSG hapten, wherein the targetable construct is conjugated to at least one diagnostic agent.

41. The method of claim 40, wherein the diagnostic agent is ¹⁸F and the method further comprises PET imaging.

49. A DNL construct comprising a first fusion protein and a second fusion protein, wherein the first fusion protein comprises an hPAM4 antibody or antigen-binding fragment thereof attached to a DDD peptide or an AD peptide and the second fusion protein comprises an AD peptide when the first fusion protein comprises a DDD peptide or a DDD peptide when the first fusion protein comprises an AD peptide.

50. The DNL construct of claim 49, wherein the DDD peptide is a DDD2 peptide (SEQ ID NO:

34) and the AD peptide is an AD2 peptide (SEQ ID NO;

36).

51. The DNL construct of claim 49, wherein the second fusion protein is conjugated to a polyethylene glycol (PEG) moiety.

52. The DNL construct of claim 49, wherein the second fusion protein comprises a cytokine moiety.

53. The DNL construct of claim 52, wherein the cytokine moiety is interferon-α

2b.

54. The DNL construct of claim 49, wherein the second fusion protein comprises an h679 antibody or antigen-binding fragment thereof.

55. Use of a DNL construct according to claim 49 for the manufacture of a medicament for the therapy, detection or diagnosis of pancreatic cancer.