Bispecific antibody point mutations for enhancing rate of clearance

• US 20040018557A1
• Filed: 03/03/2003
• Published: 01/29/2004
• Est. Priority Date: 03/01/2002
• Status: Abandoned Application

• Current Assignee: Immunomedics, Inc.
• Original Assignee: Immunomedics, Inc.
• Inventors: Qu, Zhengxing, Hansen, Hans, Goldenberg, David
• Application Number:
  US10/377,109
• Publication Number:
  US 20040018557A1
• Time in Patent Office: Days
• US Class Current: 435/7.1
• CPC Class Codes: A61K 2039/505 comprising 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 51/109 immunoglobulins having two or more different antigen-binding sites or multifunctional antibodies
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 33/00 : Antiparasitic agents
• CPC Class Codes: A61P 33/02 : Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 5/00 : Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: B82Y 5/00 : Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 16/3007 : Carcino-embryonic Antigens
• CPC Class Codes: C07K 16/468 : Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• 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 2317/52 : Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/524 : CH2 domain
• CPC Class Codes: C07K 2317/53 : Hinge
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance

Abstract

A mutant bispecific antibody that includes (a) a human hinge constant region from IgG having one or more amino acid mutations in the C_H2 domain, (b) two scFvs; and (c) two Fvs has been constructed. This type of antibody displays enhanced clearance, which has been found to be particularly useful in the context of pre-targeting methods.

Claims

1. A mutant bispecific antibody, comprising at least:

(a) a human hinge constant region from IgG;

(b) two scFvs; and

(c) two Fvs,

wherein said constant region contains one or more amino acid mutations in the C_H2 domain.

2. The mutant bispecific antibody of claim 1, wherein said scFvs and said Fvs are CDR-grafted murine scFvs and Fvs.

3. The mutant bispecific antibody of claim 1, wherein said scFvs and said Fvs are humanized or human.

4. The mutant bispecific antibody of claim 1, wherein said hinge constant region contains a mutation in which isoleucine at position 253 is replaced with alanine, or amino acids other than leucine, wherein the amino acid replacement enhances blood clearance equal to, or greater than replacement with alanine.

5. The mutant bispecific antibody of claim 4, wherein said Fvs are derived from hMN14-IgG and said scFvs are 734scFv.

6. The mutant bispecific antibody of claim 1, wherein said scFvs bind a monovalent targetable construct.

7. The mutant bispecific antibody of claim 1, wherein said scFvs bind a divalent targetable construct.

8. The mutant bispecific antibody of claim 1, wherein said Fvs bind to an epitope on a target cell.

9. A method of treating or diagnosing or treating and diagnosing a disease or a condition that may lead to a disease comprising

(A) administering to said subject the mutant bispecific antibody of claim 1, wherein the Fvs are directed to a marker substance associated with the disease or condition;

(B) optionally, administering to said subject a clearing composition, and allowing said composition to clear non-localized antibodies or antibody fragments from circulation; and

(C) administering to said subject a targetable construct comprising a bivalent hapten, wherein both hapten moieties bind to the two scFvs on a single molecule of the mutant bi-specific of claim 1, wherein the targetable construct further comprises a diagnostic or therapeutic cation, and/or one or more chelated or chemically bound therapeutic or diagnostic agents.

10. The method as claimed in claim 9, wherein said mutant bispecific antibody is administered before, during or after the administration of at least one therapeutic agent used to treat the disease or condition.

11. The method as claimed in claim 9, wherein said targetable construct comprises an enzyme and said method further comprises administering to said subject

a) a prodrug, when said enzyme is capable of converting said prodrug to a drug at the target site;

or

b) a drug which is capable of being detoxified in said subject to form an intermediate of lower toxicity, when said enzyme is capable of reconverting said detoxified intermediate to a toxic form, and, therefore, of increasing the toxicity of said drug at the target site, or

c) a prodrug which is activated in said subject through natural processes and is subject to detoxification by conversion to an intermediate of lower toxicity, when said enzyme is capable of reconverting said detoxified intermediate to a toxic form, and, therefore, of increasing the toxicity of said drug at the target site.

12. The method of claim 11, wherein said prodrug is selected from the group consisting of epirubicin glucuronide, CPT-11, etoposide glucuronide, daunomicin glucuronide and doxorubicin glucuronide

13. The method of claim 9, wherein said diagnostic agent emits 25 to 4,000 keV gamma particles and/or positrons.

14. The method of claim 9, wherein the diagnostic agent is selected from the group consisting of ¹⁸F, ⁵²Fe, ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁶⁷Ga, ⁶⁸Ga, ⁸⁶Y, ⁸⁹Zr ^94mTc, ⁹⁴Tc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, ¹²⁵I, ¹³¹I, ^154-158Gd, ¹⁷⁷Lu, ³²P, 188Re, and ⁹⁰Y or a combination thereof.

15. The method of claim 9, wherein said radioactive isotope is used to perform positron-emission tomography (PET).

16. The method of claim 9, wherein said targetable construct comprises one or more image enhancing agents for use in magnetic resonance imaging (MRI).

17. The method of claim 16, wherein said enhancing agent is selected from the group consisting of Mn, Fe and Gd.

18. The method of claim 9, wherein said targetable construct comprises one or more image enhancing agents for use in ultrasound imaging.

19. The method of claim 18, wherein said targetable construct is a liposome with a bivalent DTPA-peptide covalently attached to the outside surface of the liposome lipid membrane.

20. The method of claim 19, wherein said liposome is gas filled.

21. The method of claim 9, wherein said targetable construct comprises one or more radioactive isotopes useful for killing diseased tissue.

22. The method of claim 21, wherein the energy range of the radioactive isotope is 60 to 700 keV.

23. The method of claim 21, wherein said radioactive isotope is selected from the group consisting of ³²P, ³³P, ⁴⁷Sc, ⁶⁴Cu, ⁶⁷Cu, ⁶⁷Ga, ⁹⁰Y, ¹¹¹Ag, ₁₁₁In, ¹²⁵I, ¹³¹I, ¹⁴²Pr, ¹⁵³Sm, ¹⁶¹Tb, ¹⁶⁶Dy, ¹⁶⁶ Ho, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁸⁹Re, ²¹²Pb, ²¹²Bi, ²¹³Bi, ²¹¹At, ²²³Ra and ²²⁵Ac or a combination thereof.

24. The method of claim 21, wherein said targetable construct comprises ¹⁰B atoms, and said method further comprises the step of irradiating said boron atoms localized at said diseased tissue, thereby effecting BNCT of said diseased tissue.

25. The method of claim 9, wherein said therapeutic agent is a drug, toxin, hormone, enzyme, immunomodulator, chelator, boron compound, photoactive agent, dye, or radioisotopes.

26. The method of claim 21, wherein said targetable construct comprises one or more toxins.

27. The method of claim 26, wherein said toxin is selected from the group consisting of ricin, abrin, ribonuclease, DNase 1, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtherin toxin, Pseudomonas exotoxin, and Pseudomonas endotoxin or a combination thereof.

28. The method of claim 9, wherein said targetable construct comprises one or more drugs.

29. The method of claim 28, wherein said drug is selected from the group consisting of nitrogen mustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, triazenes, folic acid analogs, anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzymes, epipodophyllotoxins, platinum coordination complexes, vinca alkaloids, substituted ureas, methyl hydrazine derivatives, adrenocortical suppressants, antagonists, endostatin, taxols, camptothecins, doxorubicins and their analogs, and a combination thereof.

30. The method of claim 9, wherein the targetable construct comprises one or more agents for photodynamic therapy.

31. The method of claim 30, wherein said agent for photodynamic therapy is a photosensitizer.

32. The method of claim 31, wherein said photosensitizer is selected from the group consisting of benzoporphyrin monoacid ring A (BPD-MA), tin etiopurpurin (SnET2), sulfonated aluminum phthalocyanine (AlSPc) and lutetium texaphyrin (Lutex).

33. The method of claim 9, wherein said targeted tissue is a tumor.

34. The method of claim 11, wherein said targeted tissue is a tumor.

35. The method of claims 33 or 34, wherein said tumor produces or is associated with antigens selected from the group consisting of colon-specific antigen-p (CSAp), carcinoembryonic antigen (CEA), CD4, CD5, CD8, CD14, CD15, CD19, CD20, CD21, CD22, CD23, CD25, CD33, CD37, CD38, CD40, CD40L, CD46, CD52, CD54, CD66a-e, CD74, CD75, CD80, CD126, B7, HLA-DR, Ia, Ii, HM1.24, MUC 1, MUC2, MUC3, MUC4,Tag-72, PSMA, EGP-1, EGP-2, PSA, AFP, HCG, HCG-beta, PLAP, PAP, histone, tenascin, VEGF, P1GF, S10O, EGFR, insulin-like growth factor, HER2/neu, organotropic hormones, oncogene products, and cytokeratin.

36. The method of claims 9 or 11, wherein the mutant bispecific antibody incorporates the Fv of a Class III anti-CEA antibody.

37. The method of claims 9 or 11, wherein the mutant bispecific antibody incorporates the scFv of Mab 679.

38. The method of claims 9 or 11, wherein said disease is an immune dysregulation disease, an autoimmune disease, organ graft rejection, cardiovascular disease, neurological disease or graft vs. host disease.

39. The method of claim 38 wherein said autoimmune disease is selected from the group consisting of acute idiopathic thrombocytopenic purpura, chronic idiopathic thrombocytopenic purpura, dermatomyositis, Sydenham'"'"'s chorea, myasthenia gravis, systemic lupus erythematosus, lupus nephritis, rheumatic fever, polyglandular syndromes, bullous pemphigoid, diabetes mellitus, Henoch-Schonlein purpura, post-streptococcalnephritis, erythema nodosurn, Takayasu'"'"'s arteritis, Addison'"'"'s disease, rheumatoid arthritis, multiple sclerosis, sarcoidosis, ulcerative colitis, erythema multiforme, IgA nephropathy, polyarteritis nodosa, ankylosing spondylitis, Goodpasture'"'"'s syndrome, thromboangitisubiterans, Sjogren'"'"'s syndrome, primary biliary cirrhosis, Hashimoto'"'"'s thyroiditis, thyrotoxicosis, scleroderma, chronic active hepatitis, polymyositis/dermatomyositis, polychondritis, parnphigus vulgaris, Wegener'"'"'s granulomatosis, membranous nephropathy, amyotrophic lateral sclerosis, tabes dorsalis, giant cell arteritis/polymyalgia, perniciousanemia, rapidly progressive glomerulonephritis and fibrosing alveolitis.

40. The method of claims 9 or 11, wherein said disease caused by a fungus, virus, parasite or bacterium, and the Fv of the mutant bispecific targets the fungus, virus, parasite, or bacterium.

41. The method of claim 40, wherein said virus is selected from the group consisting of human immunodeficiency virus (HIV), herpes virus, cytomegalovirus, rabies virus, influenza virus, hepatitis B virus, Sendai virus, feline leukemia virus, Reo virus, polio virus, human serum parvo-like virus, simian virus 40, respiratory syncytial virus, mouse mammary tumor virus, Varicella-Zoster virus, Dengue virus, rubella virus, measles virus, adenovirus, human T-cell leukemia viruses, Epstein-Barr virus, murine leukemia virus, mumps virus, vesicular stomatitis virus, Sindbis virus, lymphocytic choriomeningitis virus, wart virus and blue tongue virus.

42. The method of claim 40, wherein said bacterium is selected from the group consisting of Anthrax bacillus, Streptococcus agalactiae, Legionella pneumophilia, Streptococcus pyogenes, Escherichia coli, Neisseria gonorrhoeae, Neisseria meningitidis, Pneumococcus, Hemophilis influenzae B, Treponema pallidum, Lyme disease spirochetes, Pseudomonas aeruginosa, Mycobacterium leprae, Brucella abortus, Mycobacterium tuberculosis and Tetanus toxin.

43. The method of claim 40, wherein said pathogen is a protozoan.

44. The method of claim 43, wherein said protozoan is selected from the group consisting of Plasmodium falciparum, Plasmodium vivax, Toxoplasma gondii, Trypanosoma rangeli, Trypanosoma cruzi, Trypanosoma rhodesiensei, Trypanosoma brucei, Schistosoma mansoni, Schistosoma japanicum, Babesia bovis, Elmeria tenella, Onchocerca volvulus, Leishmania tropica, Trichinella spiralis, Onchocerca volvulus, Theileria parva, Taenia hydatigena, Taenia ovis, Taenia saginata, Echinococcus granulosus and Mesocestoides corti.

45. The method of claim 40, wherein said parasite is a helminth or a malarial parasite.

46. The method of claim 40, wherein said bacterium is mycoplasma.

47. The method of claim 46, wherein said mycoplasma is selected from the group consisting of Mycoplasma arthritidis, M. hyorhinis, M. orale, M. arginini, Acholeplasma laidlawii, M. salivarum, and M. pneumoniae.

48. The method of claim 40, wherein the fungus is selected from the group consisting of Histoplasma capsulatum, Blastomyces dermatitidis, Coccidioides immitis, and species of Candida.

49. The method of claims 9 or 11, wherein the tissue is normal ectopic tissue.

50. The method of claim 49, wherein said normal tissue is tissue from the ovary, thymus, parathyroid, bone marrow, or spleen.

51. The method of claims 9 or 11, wherein said subject is mammalian.

52. The method of claims 9 or 11, wherein said mammalian subject is a human or primate.

53. The method of claim 51, wherein said mammalian subject is selected from the group consisting of rodents, lagamorphs, bovines, ovines, caprines, porcines, equines, canines, felines, domestic fowl, ungulates, and bear.

54. The method of claim 9, wherein the application is for intraoperative diagnosis to identify diseased tissues.

55. The method of claim 9, wherein the application is for endoscopic diagnosis to identify diseased tissues.

56. The method of any one of claims 9-55 wherein a second therapeutic agent is administered before, concurrently, or after the prescribed diagnosis or treatment.

57. The method of claim 56, wherein the second therapeutic agent is a drug, naked antibody, immunomodulator, or antibody conjugate bearing a drug, radioisotope, immunomodulator or toxin.

58. A kit useful for treating or identifying diseased tissues in a subject comprising:

(A) the mutant bispecific antibody of claim 9;

(B) optionally, the clearing agent of claim 9;

and

(C) the targetable construct of claim 11.

59. A kit useful for treating or identifying diseased tissues in a subject comprising:

(A) the mutant bispecific antibody of claim 11;

(B) optionally, the clearing agent of claim 11;

(C) the targetable construct of claim 11;

and

(D) the prodrug of claim 11.

60. The method of claim 25, 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), erythropoietin, thrombopoietin and a combination thereof.

61. The method of claim 60, wherein said lymphotoxin is tumor necrosis factor (TNF).

62. The method of claim 60, wherein hematopoietic factor is interleukin (IL).

63. The method of claim 60, wherein said colony stimulating factor is granulocyte-colony stimulating factor (G-CSF) or granulocyte macrophage-colony stimulating factor (GM-CSF).

64. The method of claim 60, wherein said interferon is interferon-α

, -β

or -γ

.

65. The method of claim 60, wherein said stem cell growth factor is S1 factor.

66. The method of claim 25, wherein said immunomodulator is IL-1, IL-2, IL-3, IL-6, IL-10, IL-12, IL-18, interferon-γ

, TNF-α

or a combination thereof.

67. The method of claim 38, wherein said neurological disease is Alzheimer'"'"'s Disease.