Cross-reacting monoclonal antibodies specific for E- and P-selectin

• US 5,622,701 A
• Filed: 06/14/1994
• Issued: 04/22/1997
• Est. Priority Date: 06/14/1994
• Status: Expired due to Term

• Current Assignee: Abbvie Biotherapeutics Inc. (Abbvie Incorporated)
• Original Assignee: Protein Design Labs, Inc. (PDL BioPharma, Inc.)
• Inventors: Berg, Ellen L.
• Primary Examiner(s): Chan, Christina Y.
• Assistant Examiner(s): Gambel, Phillip
• Application Number:
  US08/259,963
• Time in Patent Office: 1,043 Days
• Field of Search: 435/240.27, 435/172.2, 435/69.1, 435/70.1, 435/70.21, 435/69.6, 435/172.3, 435/290.27, 436/548, 436/536, 424/133.1, 424/136.1, 424/152.1, 424/143.1, 424/172.1, 424/144.1, 424/130.1, 424/141.1, 424/1, 424/173.1, 424/184.1, 424/153.1, 514/885, 530/387.3, 530/387.1, 530/388.1, 530/380.22, 530/388.7, 530/388.73, 530/389.6, 536/23.53, 536/23.5, 536/23.57
• US Class Current: 424/153.1
• 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 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• 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 31/04 Antibacterial 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 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/08 Vasodilators for multiple indications
• 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: C07K 16/2854 against selectins, e.g. CD62
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

The invention provides monoclonal antibodies that specifically bind to P-selectin and to E-selectin. Many of the antibodies block the functional interactions of P-selectin and E-selectin with the irrespective counterreceptors.

Claims

1. A humanized monoclonal antibody having a binding site that specifically binds to P-selectin and to E-selectin comprising a humanized heavy chain and a humanized light chain:

(1) the humanized light chain comprising three complementarily determining regions (CDR1, CDR2 and CDR3) having amino acid sequences from the mouse antibody light chain and comprising a variable region framework sequence substantially identical to a human light chain variable region framework sequence; and

(2) the humanized heavy chain comprising three complementarily determining regions (CDR1, CDR2 and CDR3 having amino acid sequences from the mouse antibody heavy chain, and comprising a variable region framework sequence substantially identical to a human heavy chain variable region framework sequence;

wherein the humanized antibody specifically binds to the P-selectin with a binding affinity having a lower limit of 10⁷ M^-1 ;

wherein the humanized antibody specifically binds to the E-selectin with a binding affinity having a lower limit of 10⁷ M^-1 ;

and wherein the specific binding of the antibody to the P-selectin inhibits binding of the P-selectin to a counter-receptor of P-selectin; and

the specific binding of the antibody to the E-selectin inhibits binding of the E-selectin to a counter-receptor of E-selectin.

2. The antibody of claim 1 that does not specifically bind to L-selectin.

3. The antibody of claim 2 that is a Fab, Fab'"'"', F (ab'"'"')₂, Fv fragment, or a single-chain antibody.

4. The antibody of claim 2 wherein the complementarily determining regions (CDR1, CDR2 and CDR3 of the light and heavy chains have amino acid sequences from the light and heavy chains of monoclonal antibody 5C7.29, ATCC accession number CRL 11640, respectively.

6. A purified nucleic acid segment encoding a light or heavy chain variable region of the antibody of claim 2.

7. A stable cell line comprising:

a nucleic acid segment encoding the heavy chain of the antibody of claim 2, the segment operably linked to a first

promoter to allow expression of the heavy chain;

a second nucleic acid segment encoding the light chain of the antibody of claim 2, the second segment operably linked to a second promoter to allow expression of the light chain;

wherein the stable cell line can produce the antibody of claim 2.

8. A pharmaceutical composition comprising a therapeutically effective amount of the monoclonal antibody of claim 2 in an acceptable carrier.

9. A method of treating an inflammatory disease or condition, comprising administering to a human patient a therapeutically effective dose of the pharmaceutical composition of claim 8.

10. The method according to claim 9, wherein the inflammatory disease is ischemia-reperfusion injury.

11. The method of claim 10, wherein the inflammatory disease is ischemia-reperfusion injury after myocardial infarction.

5. An antibody made by hybridoma 5C7.29, ATCC accession number CRL 11640.

12. A method of generating an antibody capable of blocking E-selectin and P-selectin mediated functions, but not L-selectin mediated functions, the method comprising:

immunizing a mammal with P-selectin;

immunizing the mammal with E-selectin;

immortalizing B-cells from the mammal to obtain immortalized cells producing antibodies; and

selecting an immortalized cell producing an antibody that specifically binds to E-selectin and to P-selectin, but does not specifically bind to L-selectin, and whose specific binding to P-selectin inhibits binding of P-selectin to a counter-receptor of P-selectin; and

whose specific binding of the antibody to E-selectin inhibits binding of E-selectin to a counter-receptor of E-selectin.