Human poly (ADP-ribose) polymerase 2 materials and methods

• US 20030170859A1
• Filed: 02/19/2003
• Published: 09/11/2003
• Est. Priority Date: 06/16/1999
• Status: Active Grant

• Current Assignee: Anthony J. Demaggio, David L. Mcelligott, Erik Christenson, Phyllis S. Goldman
• Original Assignee: Anthony J. Demaggio, David L. Mcelligott, Erik Christenson, Phyllis S. Goldman
• Inventors: Christenson, Erik, Goldman, Phyllis S., McElligott, David L., Demaggio, Anthony J.
• Granted Patent: US 6,989,260 B2
• Time in Patent Office: Days
• US Class Current: 435/199
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 11/08 Bronchodilators
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 31/04 Antibacterial agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 9/1077 : Pentosyltransferases (2.4.2)
• CPC Class Codes: C12Y 204/0203 : NAD+ ADP-ribosyltransferase (2.4.2.30), i.e. tankyrase or poly(ADP-ribose) polymerase

Abstract

The invention provides a novel human poly(ADP-ribose) polymerase (hPARP2) polypeptides, polynucleotides encoding the polypeptides, expression constructs comprising the polynucleotides, and host cells transformed with the expression constructs. Also provided are methods for producing the hPARP2 polypeptides, antibodies that are immunoreactive with the hPARP2 polypeptides. In addition, there are provided methods for identifying specific binding partners of hPARP2, and more particularly methods for identifying binding partners that modulate biological activity of hPARP2. Methods of modulating biological activity of hPARP2 in vitro and in vivo are also provided.

Claims

1. A purified and isolated hPARP2 polypeptide, comprising the amino acid sequence defined in SEQ ID NO:

2 or a functional derivative thereof.

2. A polynucleotide encoding the hPARP2 polypeptide according to claim 1.

3. The polynucleotide according to claim 2, comprising the nucleotide sequence defined in SEQ ID NO:

1.

4. A polynucleotide encoding a hPARP2 polypeptide selected from the group consisting of:

a) the polynucleotide according to claim 2,

b) the polynucleotide according to claim 3, and

c) a polynucleotide that hybridizes under moderately stringent hybridization conditions to the complement of the polynucleotide of (a) or (b).

5. An hPARP2 polypeptide encoded by the polynucleotide according to claim 4.

6. The polynucleotide according to claim 4, which is a DNA molecule or an RNA molecule.

7. The polynucleotide according to claim 6, which further comprises a detectable label moiety.

8. An expression construct, comprising the polynucleotide according to claim 4.

9. A host cell transformed or transfected with the expression construct according to claim 8.

10. The polynucleotide according to claim 4, wherein the polynucleotide is operatively linked to a heterologous promoter.

11. A host cell, comprising the polynucleotide according to claim 10.

12. A method for producing a polypeptide having an amino acid sequence defined by SEQ ID NO:

2, comprising the steps of;

a) growing the host cell according to claim 9 or 11 under conditions appropriate for expression of the polypeptide; and

b) isolating the polypeptide from the host cell or the medium in which the host cell is grown.

13. An antibody that is specifically immunoreactive with the polypeptide according to claim 1.

14. The antibody according to claim 13, wherein the antibody is selected from the group consisting of monoclonal antibodies, polyclonal antibodies, single chain antibodies (scFv antibodies), chimeric antibodies, multifunctional/multispecific antibodies, humanized antibodies, human antibodies, CDR-grafted antibodies, Fab fragments, Fab′

fragments, F(ab′

)₂ fragments, Fv fragments, diabodies;

linear antibodies;

single-chain antibody molecules; and

multispecific antibodies formed from antibody fragments.

15. A cell line that produces the antibody according to claim 13.

16. An anti-idiotype antibody that is specifically immunoreactive with an antibody according to claim 14.

17. A method for identifying a specific binding partner of the hPARP2 polypeptide according to claim 1, comprising:

a) contacting the hPARP2 polypeptide with a test compound under conditions that permit binding of the hPARP2 polypeptide and the test compound;

b) detecting binding of the test compound and the hPARP2 polypeptide; and

c) identifying the test compound as a specific binding partner of the hPARP2 polypeptide.

18. The method according to claim 17, wherein said specific binding partner modulates a biological activity of the hPARP2 polypeptide.

19. The method according to claim 18, wherein said specific binding partner inhibits a biological activity of the hPARP2 polypeptide.

20. The method according to claim 18, wherein said specific binding partner enhances a biological activity of the hPARP2 polypeptide.

21. A method for identifying a specific binding partner of the hparp2 polynucleotide according to claim 2, comprising:

a) contacting the hparp2 polynucleotide with a test compound under conditions that permit binding of the hparp2 polynucleotide and the test compound;

b) detecting binding of the test compound and the hparp2 polynucleotide; and

c) identifying the test compound as a specific binding partner of the hparp2 polynucleotide.

22. The method according to claim 21, wherein said specific binding partner modulates expression of the hPARP2 polypeptide.

23. The method according to claim 22, wherein said specific binding partner inhibits expression of the hPARP2 polypeptide.

24. A method according to claim 22, wherein said specific binding partner enhances expression of the hPARP2 polypeptide.

25. A method of treating a human subject having a disorder mediated by poly(ADP-ribose) polymerase activity, comprising administering to said subject an hPARP2 antagonist in an amount effective for inhibiting expression or activity of the polypeptide according to claim 1.

26. The method according to claim 25, wherein the hPARP1 antagonist inhibits hPARP1.

27. The method according to claim 25, wherein said disorder is selected from the group consisting of inflammatory disorders, neurological disorders, cardiovascular disorders, and disorders of neoplastic tissue growth.

28. The method according to claim 27, wherein said disorder is an inflammatory disorder.

29. The method according to claim 25, wherein said disorder is characterized by reperfusion injury.

30. The method according to claim 29, wherein said disorder is selected from the group consisting of ischemic stroke, hemorrhagic shock, myocardial ischemia or infarction, transplantation, and cerebral vasospasm.

31. The method according to claim 28, wherein said disorder is selected from the group consisting of rheumatoid arthritis, osteoarthritis, gouty arthritis, spondylitis;

Behcet disease;

sepsis, septic shock, endotoxic shock, gram negative sepsis, gram positive sepsis, toxic shock syndrome;

multiple organ injury syndrome secondary to septicemia, trauma, or hemorrhage;

allergic conjunctivitis, vernal conjunctivitis, uveitis, thyroid-associated ophthalmopathy;

cosinophilic granuloma;

asthma, chronic bronchitis, allergic rhinitis, ARDS, chronic obstructive pulmonary disease, silicosis, pulmonary sarcoidosis, pleurisy, alveolitis, vasculitis, pneumonia, bronchiectasis, pulmonary oxygen toxicity;

reperfusion injury of the myocardium, brain, or extremities;

cystic fibrosis;

keloid formation, scar tissue formation;

atherosclerosis;

systemic lupus erythematosus, autoimmune thyroiditis, multiple sclerosis;

Reynaud'"'"'s syndrome;

graft versus host disease, allograft rejection;

chronic glomerulonephritis;

inflammatory bowel disease, Crohn'"'"'s disease, ulcerative colitis, necrotizing enterocolitis;

inflammatory dermatoses, contact dermatitis, atopic dermatitis, psoriasis, urticaria, fever and myalgias due to infection;

meningitis, encephalitis, and brain or spinal cord injury due to minor trauma;

Sjö

gren'"'"'s syndrome;

diseases involving leukocyte diapedesis;

alcoholic hepatitis;

bacterial pneumonia;

antigen-antibody complex mediated diseases;

hypovolemic shock;

Type I diabetes mellitus;

acute and delayed hypersensitivity;

disease states due to leukocyte dyscrasia and metastasis;

thermal injury;

granulocyte transfusion associated syndromes; and

cytokine-induced toxicity.

32. A method of inhibiting poly(ADP-ribose) polymerase activity in a cell, comprising contacting said cell with an hPARP2 antagonist in an amount effective for inhibiting expression or activity of the polypeptide according to claim 1.