Antibody fragment-peg conjugates

• US 7,005,504 B2
• Filed: 11/29/2000
• Issued: 02/28/2006
• Est. Priority Date: 01/22/1998
• Status: Expired due to Fees

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Shahrokh, Zahra, Leong, Steven, Hsei, Vanessa, Presta, Leonard, Koumenis, Iphigenia, Zapata, Gerardo
• Primary Examiner(s): HELMS, LARRY RONALD
• Application Number:
  US09/726,258
• Publication Number:
  US 20030021790A1
• Time in Patent Office: 1,917 Days
• Field of Search: 530/391.1, 530/391.3, 530/391.5, 530/388.23, 530/350, 424/130.1, 424/192.1, 424/153.1, 424/194.1, 424/178.1, 424/139.1
• US Class Current: 530/388.23
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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: 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 2317/565 Complementarity determining region [CDR]
• 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 comprising a humanized antibody Fab′

fragment further comprising an antigen binding site,

wherein said antigen binding site (a) binds to human interleukin-8 (IL-8), and (b) comprises the complementarity determining regions of (i) 6G4V11N35A light chain (FIG. 36, SEQ ID NO;

     56) or (ii) the 6G4V11N35E light chain (FIG. 45, SEQ ID NO;

     62),

wherein said humanized antibody Fab′

fragment is covalently attached to a polyethylene glycol (PEG) molecule having an average molecular weight of at least 20 kD, and

wherein the apparent size of the conjugate is at least about 500 kD, and at least about 8 fold greater than the apparent size of the antibody fragment.

2. The conjugate of claim 1 wherein in the PEG molecule is covalently attached to the hinge region of the antibody fragment.

3. The conjugate of claim 1 wherein said antigen binding site comprises the complementarity determining regions of FIG. 36 (SEQ ID NO:

56).

4. The conjugate of claim 1, wherein the covalent structure of the conjugate is free of any matter other than the antibody fragment and PEG that form the conjugate.

5. The conjugate of claim 1, wherein the covalent structure of the conjugate incorporates one or more nonproteinaceous labels, and wherein the covalent structure of the conjugate is free of any matter other than the antibody fragment, PEG and nonproteinaceous label molecules that form the conjugate.

6. The conjugate of claim 5 wherein at least one nonproteinaceous label is radiolabel.

9. A composition comprising the conjugate of claim 3 and a carrier.

10. The composition of claim 9 that is sterile.

11. A composition comprising the conjugate of claim 1 and a carrier.

12. The composition of claim 11 that is sterile.

13. The conjugate of claim 1 wherein said antigen binding site comprises the complementarity determining regions of FIG. 45 (SEQ ID NO:

62).

14. A composition comprising the conjugate of claim 13 and a carrier.

15. The composition of claim 14 that is sterile.

7. A conjugate comprising a humanized antibody Fab′

fragment,

wherein said antibody fragment;

(a) binds to human interleukin 8 (IL-8);

(b) comprises the complementarity determining regions of (i) 6G4V11N35A light chain (FIG. 36, SEQ ID NO;

     56) or (ii) 6G4V11N35E light chain (FIG. 45, SEQ ID NO;

     62), and (c) is covalently attached to a single polyethylene glycol (PEG) molecule of at least 20 kD, and

wherein said conjugate has an apparent size of at least about 500 kD and at least about 8 fold greater than the apparent size of the antibody fragment.

8. A conjugate comprising a humanized antibody Fab′

fragment,

wherein said antibody fragment;

(a) binds to human interleukin 8 (IL-8);

(b) comprises the complementarity determining regions of (i) 6G4V11N35A light chain (FIG. 36, SEQ ID NO;

     56) or (ii) 6G4V11N35E light chain (FIG. 45, SEQ ID NO;

     62), and (c) is covalently attached to a polyethylene glycol (PEG) molecule having an average molecular weight of at least about 40 kD,

wherein the apparent size of the conjugate is at least about 500 kD, and at least about 8 fold greater than the apparent size of the antibody fragment.