Antibody fragment-polymer conjugates and humanized anti-IL-8 monoclonal antibodies

• US 6,870,033 B1
• Filed: 02/20/1998
• Issued: 03/22/2005
• Est. Priority Date: 02/21/1997
• Status: Expired due to Fees

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Zapata, Gerarado A., Shahrokh, Zahra, Presta, Leonard G., Koumenis, Iphigenia, Hsei, Vanessa, Leong, Steven R.
• Primary Examiner(s): HELMS, LARRY RONALD
• Application Number:
  US09/355,014
• Time in Patent Office: 2,587 Days
• Field of Search: 530/387.1, 530/387.3, 530/388.85, 530/391.5, 424/130.1, 424/133.1
• US Class Current: 530/388.85
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/6845 the antibody targeting a cytokine, e.g. growth factors, VEGF, TNF, a lymphokine or an interferon
• CPC Class Codes: A61K 47/6883 Polymer-drug antibody conjugates, e.g. mitomycin-dextran-Ab; DNA-polylysine-antibody complex or conjugate used for therapy
• 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 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C07K 16/244 Interleukins [IL]
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

Humanized anti-IL-8 monoclonal antibodies and variants thereof are described for use in diagnostic applications and in the treatment of inflammatory disorders. Also described is a conjugate formed by an antibody fragment covalently attached to a non-proteinaceous polymer, wherein the apparent size of the conjugate is at least about 500 kD. The conjugate exhibits substantially improved half-life, mean residence time, and/or clearance rate in circulation as compared to the underivatized parental antibody fragment.

Claims

1. A conjugate consisting essentially of an antibody fragment covalently modified by one or two nonproteinaceous polymer molecules at a free sulfhydryl group of a cysteine residue within the hinge region of the antibody fragment, wherein (a) the apparent molecular weight of the conjugate, as determined by size exclusion chromatography, is at least about 500 kD, (b) the average actual molecular weight of each nonproteinaceous molecule is at least 20 kD, and (c) the antibody fragment has an antigen binding site that binds human interleukin-8 (IL-8) comprising an anti-IL-8 monoclonal antibody light chain sequence comprising amino acids 1-219 of the humanized antibody light chain 6G4.2.5v11 (SEQ ID NO:

51).

2. The conjugate of claim 1, wherein the antibody fragment is selected from the group consisting of Fab, Fab′

, and Fab′

-SH modified by one PEG molecule having an average actual molecular weight of about 30 kD.

3. The conjugate of claim 1, wherein the antibody fragment comprises an anti-IL-8 monoclonal antibody light chain sequence comprising the amino acid sequence of humanized antibody light chain 6G4.2.5v11 (SEQ ID NO:

51).

4. The conjugate of claim 3, wherein the antibody fragment is selected from the group consisting of Fab, Fab′

, and Fab′

-SH modified by one PEG molecule having an average actual molecular weight of about 30 kD.

5. A conjugate consisting essentially of an antibody fragment covalently modified by one or two nonproteinaceous polymer molecules at a free sulfhydryl group of a cysteine residue within the hinge region of the antibody fragment, wherein (a) the apparent molecular weight of the conjugate, as determined by size exclusion chromatography, is at least about 500 kD, (b) the average actual molecular weight of each nonproteinaceous molecule is at least 20 kD, and (c) the antibody fragment has an antigen binding site that binds human interleukin-8 (IL-8) comprising an anti-IL-8 monoclonal antibody heavy chain sequence comprising amino acids 1-230 of the humanized antibody 6G4.2.5v11 heavy chain (SEQ ID NO:

60).

6. The conjugate of claim 5, wherein the antibody fragment is selected from the group consisting of Fab, Fab′

, and Fab′

-SH modified by one PEG molecule having an average actual molecular weight of about 30 kD.

7. The conjugate of claim 5, further comprising a leucine zipper sequence.

8. The conjugate of claim 5, wherein the antibody fragment comprises an anti-IL-8 monoclonal antibody heavy chain sequence comprising the amino acid sequence of the humanized antibody 6G4.2.5v11 heavy chain (SEQ ID NO:

60).

9. The conjugate of claim 8, wherein the antibody fragment is selected from the group consisting of Fab, Fab′

, and Fab′

-SH modified by one PEG molecule having an average actual molecular weight of about 30 kD.

10. The conjugate of claim 8, further comprising a leucine zipper sequence.