Monoclonal antibody hPAM4

• US 7,282,567 B2
• Filed: 06/16/2003
• Issued: 10/16/2007
• Est. Priority Date: 06/14/2002
• Status: Expired due to Fees

• Current Assignee: Immunomedics, Inc. (Gilead Sciences Inc.)
• Original Assignee: Immunomedics, Inc. (Gilead Sciences Inc.)
• Inventors: Qu, Zhengxing, Hansen, Hans J., Goldenberg, David M.
• Primary Examiner(s): Helms; Larry R.
• Assistant Examiner(s): Tungaturthi; Parithosh K.
• Application Number:
  US10/461,885
• Publication Number:
  US 20050014207A1
• Time in Patent Office: 1,583 Days
• Field of Search: 530/387.3, 530/350, 530/380, 530/385, 530/387.1, 530/387.7, 530/388.1, 424/134.1, 424/135.1, 424/141.1, 424/155.1, 424/142.1, 424/178.1, 424/85.1, 435/7.9, 435/188, 435/320.1, 435/54.1
• US Class Current: 530/387.3
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 47/68 the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
• CPC Class Codes: A61K 49/0008 Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
• CPC Class Codes: A61K 51/1057 the tumor cell being from liver or pancreas
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4748 Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered

Abstract

This invention relates to monovalent and multivalent, monospecific antibodies and to multivalent, multispecific antibodies. One embodiment of these antibodies has one or more identical binding sites where each binding site binds with a target antigen or an epitope on a target antigen. Another embodiment of these antibodies has two or more binding sites where these binding sites have affinity towards different epitopes on a target antigen or different target antigens, or have affinity towards a target antigen and a hapten. The present invention further relates to recombinant vectors useful for the expression of these functional antibodies in a host. More specifically, the present invention relates to the tumor-associated antibody designated PAM4. The invention further relates to humanized and human PAM4 antibodies, and the use of such antibodies in diagnosis and therapy.

Claims

1. A humanized antibody or an antigen binding fragment thereof, comprising the complementarity-determining regions (CDRs) of a murine PAM4 monoclonal antibody (MAb) and the framework (FR) regions of the light and heavy chain variable regions of a human antibody and the light and heavy chain constant regions of a human antibody, wherein the CDRs of the light chain variable region of the humanized PAM4 Mab comprise CDR1 (SASSSVSSSYLY, SEQ ID NO:

1);

CDR2 (STSNLAS, SEQ ID NO;

2); and

CDR3 (HQWNRYPYT, SEQ ID NO;

3); and

the CDRs of the heavy chain variable region of the humanized PAM4 Mab comprise CDR1 (SYVLH, SEQ ID NO;

4);

CDR2 (YINPYNDGTQYNEKFKG, SEQ ID NO;

5)and CDR3 (GFGGSYGFAY, SEQ ID NO;

6), wherein said antibody or fragment thereof binds to a domain located between the amino terminus and the start of a repeat domain of MUC1.

2. The humanized antibody or fragment thereof of claim 1, wherein the FRs of the light and heavy chain variable regions of said humanized antibody or fragment thereof comprise at least one amino acid substituted from the corresponding FRs of a murine PAM4 Mab.

3. The humanized antibody or fragment thereof of claim 2, wherein said substituted amino acid from said murine PAM4 Mab is at least one amino acid selected from the group consisting of amino acid residue 5, 27, 30, 38, 48, 66, 67, and 69 of the murine heavy chain variable region of FIG. 1B, PAM4 VH amino acid sequence (SEQ ID NO:

11).

4. The humanized antibody or fragment thereof of claim 2, wherein said amino acid from said murine Mab is at least one amino acid selected from the group consisting of amino acid residue 21, 47, 59, 60, 85, 87, and 100 of the murine light chain variable region FIG. 1A, PAM4V_k sequence (SEQ ID NO:

9).

5. The antibody or fragment thereof of claim 1, wherein said antibody or fragment thereof comprises a hPAM4 V_H amino acid sequence (SEQ ID NO:

16) of FIG. 4A and a hPAM4 V_k amino acid sequence (SEQ ID NO;

     19) of FIG. 4B.

6. A cancer cell targeting diagnostic or therapeutic conjugate comprising an antibody component that comprises an antibody or fragment thereof of claim 1, wherein said antibody component is bound to at least one diagnostic and/or therapeutic agent.

7. The diagnostic conjugate according to claim 6, wherein said diagnostic/detection agent is selected from the group comprising a radionuclide, a contrast agent, and a photoactive diagnostic/detection agent.

8. The diagnostic conjugate of claim 7, wherein said diagnostic/detection agent is a radionuclide.

9. The diagnostic conjugate of claim 8, wherein said radionuclide has an energy between 20 and 4,000 keV.

10. The diagnostic conjugate of claim 8, wherein said radionuclide is a gamma-, beta- or a positron-emitting isotope.

11. The diagnostic conjugate of claim 10, wherein said radionuclide is selected from the group consisting of ¹¹⁰In, ¹¹¹In, ¹⁷⁷Lu, ¹⁸F, ⁵²Fe, ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, 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.

12. The diagnostic conjugate of claim 7, wherein said diagnostic/detection agent is a radiological contrast agent.

13. The diagnostic conjugate of claim 12, wherein said contrast agent is a paramagnetic ion.

14. The diagnostic conjugate of claim 13, wherein said paramagnetic ion is a metal comprising 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) or erbium (III).

15. The diagnostic conjugate of claim 12, wherein said contrast agent is a metal comprising lanthanum (III), gold (III), lead (II), or bismuth (III).

16. The diagnostic conjugate of claim 7, wherein said diagnostic/detection agent is a photoactive diagnostic/detection agent.

17. The diagnostic conjugate of claim 16, wherein said photoactive diagnostic/detection agent is a fluorescent labeling compound selected from the group comprising fluorescein isothiocyanate, rhodamine, phycoerytherin, phycocyanin, allophycocyanin, o-phthaldehyde and fluorescamine.

18. The diagnostic conjugate of claim 16, wherein said photoactive diagnostic/detection agent is a chemiluminescent labeling compound selected from the group comprising luminol, isoluminol, an aromatic acridinium ester, an imidazole, an acridinium salt and an oxalate ester.

19. The diagnostic conjugate of claim 16, wherein said photoactive diagnostic/detection agent is a bioluminescent compound selected from the group comprising luciferin, luciferase and aequorin.

20. The therapeutic conjugate of claim 6, wherein said therapeutic agent is selected from the group consisting of a radionuclide, an immunomodulator, a hormone, a hormone antagonist, an oligonucleotide, an enzyme, an enzyme inhibitor, a photoactive therapeutic agent, a cytotoxic agent, an angiogenesis inhibitor, and a combination thereof.

21. The therapeutic conjugate of claim 20, wherein said oligonucleotide is an antisense oligonucleotide.

22. The therapeutic conjugate of claim 21, wherein said oligonucleotide is an antisense oligonucleotide against an oncogene.

23. The therapeutic conjugate of claim 22, wherein said oncogene is bcl-2 or p53.

24. The therapeutic conjugate of claim 20, wherein said therapeutic agent is a cytotoxic agent.

25. The therapeutic conjugate of claim 24, wherein said cytotoxic agent is a drug or a toxin.

26. The therapeutic conjugate of claim 25, wherein said drug possesses the pharmaceutical property selected from the group consisting of antimitotic, alkylating, antimetabolite, antiangiogenic, apoptotic, alkaloid, and antibiotic agents and combinations thereof.

27. The therapeutic conjugate of claim 25, wherein said drug is selected from the group consisting of nitrogen mustards, gemcitabine, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, triazenes, folic acid analogs, anthracyclines, taxanes, SN-38, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzymes, enzyme inhibitors, epipodophyllotoxins, platinum coordination complexes, Vinco alkaloids, substituted ureas, methyl hydrazine derivatives, adrenocortical suppressants, hormone antagonists, endostatin, taxols, camptothecins, doxorubicins and their analogs, antimetabolites, alkylating agents, antimitotics, antiangiogenic, apoptotic agents, methotrexate, CPT-11, and a combination thereof.

28. The therapeutic conjugate of claim 25, wherein said toxin is derived from a source selected from the group comprising an animal, a plant, and a microbial source.

29. The therapeutic conjugate of claim 25, wherein said toxin is selected from the group consisting of ricin, abrin, alpha toxin, saporin, ribonuclease (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtherin toxin, Pseudomonas exotoxin, and Pseudomonas endotoxin.

30. The therapeutic conjugate of claim 20, wherein said therapeutic agent is an immunomodulator.

31. The therapeutic conjugate of claim 30, wherein said 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 interferon (IFN), a stem cell growth factor, erythropoietin, thrombopoletin and a combination thereof.

32. The therapeutic conjugate of claim 31, wherein said lymphotoxin is tumor necrosis factor (TNF), said hematopoietic factor is an interleukin (IL), said colony stimulating factor is granulocyte-colony stimulating factor (G-CSF) or granulocyte macrophage-colony stimulating factor (GM-CSF)), said interferon is interferons-alpha, -beta or -gamma, and said stem cell growth factor is designated “

S1 factor”

.

33. The therapeutic conjugate of claim 30, wherein said immunomodulator comprises IL-1, IL-2, IL-3, IL-6, IL-10, IL-12, IL-18, IL-21, interferon-gamma, TNF-alpha or a combination thereof.

34. The therapeutic conjugate of claim 20, wherein said therapeutic agent is a radionuclide.

35. The therapeutic conjugate of claim 34, wherein said radionuclide has an energy between 60 and 700 keV.

36. The therapeutic conjugate of claim 35, wherein said radionuclide is selected from the group consisting of ³²P, ³³P, ⁴⁷Sc, ⁶⁴Cu, ⁶⁷Cu, ⁶⁷Ga, ⁸⁶Y, ⁹⁰Y, ¹¹¹Ag, ¹¹¹In, ¹²⁵I, ¹³¹I, ¹⁴²Pr, ¹⁵³Sm, ¹⁶¹Tb, ¹⁶⁶Dy, ¹⁶⁶Ho, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁸⁹Re, ²¹²Pb, ²¹²Bi, ²¹³Bi, ²¹¹At, ²²³Ra and ²²⁵Ac, and combinations thereof.

37. The therapeutic conjugate of claim 20, wherein said therapeutic agent is a photoactive therapeutic agent.

38. The therapeutic conjugate of claim 37, wherein said photoactive therapeutic agent is selected from the group comprising chromogens and dyes.

39. The therapeutic conjugate of claim 20, wherein said therapeutic agent is an enzyme.

40. The therapeutic conjugate of claim 39, wherein said enzyme is selected from the group comprising malate dehydrogenase, staphylococcal nuclease, delta-V-steroid isomerase, yeast alcohol dehydrogenase, alpha-glycerophosphate dehydrogenase, triose phosphate isomerase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, beta-galactosidase, ribonuclease, urease, catalase, glucose-6-phosphate dehydrogenase, glucoamylase and acetyicholinesterase.

41. A multivalent, multispecific antibody or fragment thereof comprising at least one humanized PAM4 antibody or fragment thereof according to claim 1 and one or more hapten binding sites having affinity towards hapten molecules.

42. The antibody or fragment thereof of claim 41, further comprising a diagnostic or therapeutic agent.

43. An antibody fusion protein or fragment thereof comprising at least two humanized PAM4 Mabs or fragments thereof according to claim 1.