Recombinant tumor specific antibody and use thereof

• US 20030157054A1
• Filed: 05/03/2002
• Published: 08/21/2003
• Est. Priority Date: 05/03/2001
• Status: Active Grant

• Current Assignee: Merck Patent GmbH (Merck & Co., Inc.)
• Original Assignee: Lexigen Pharmaceuticals Corporation
• Inventors: Gillies, Stephen D., Lo, Kin-Ming, Qian, Susan X.
• Granted Patent: US 6,969,517 B2
• Time in Patent Office: Days
• US Class Current: 424/85.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/46 Hybrid immunoglobulins hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00
• 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
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

The invention provides a family of antibodies that specifically bind the human epithelial cell adhesion molecule. The antibodies comprise modified variable regions, more specially, modified framework regions, which reduce their immunogenicity when administered to a human. The antibodies, when coupled to the appropriate moiety, may be used in the diagnosis, prognosis and treatment of cancer.

Claims

1. A recombinant anti-EpCAM antibody, wherein the antibody comprises an amino acid sequence defining an immunoglobulin light chain framework region selected from the group consisting of:

(i) amino acid residues 1-23 of SEQ ID NO;

5, wherein Xaa1 is Q or E, Xaa3 is L or V, Xaa10 is I or T, Xaa11 is M or L, Xaa13 is A or L, Xaa18 is K or R, or Xaa21is M or L, provided that at least one of the amino acid residues at positions Xaa1, Xaa3, Xaa10, Xaa11, Xaa13, Xaa18, or Xaa21 is not the same as the amino acid at the corresponding position in SEQ ID NO;

1;

(ii) amino acid residues 34-48 of SEQ ID NO;

5, wherein Xaa41 is S or Q, Xaa42 is S or A, Xaa45 is P or L, or Xaa46 is W or L, provided that at least one of the amino acid residues at positions Xaa41, Xaa42, Xaa45, or Xaa46 is not the same as the amino acid at the corresponding position in SEQ ID NO;

1; and

(iii) amino acid residues 56-87 of SEQ ID NO;

5, wherein Xaa57 is F or I, Xaa69 is S or D, Xaa71 is S or T, Xaa73 is I or T, Xaa77 is M or L, Xaa79 is A or P, Xaa82 is A or F, or Xaa84 is T or V, provided that at least one of the amino acid residues at positions Xaa57, Xaa69, Xaa71, Xaa73, Xaa77, Xaa79, Xaa82, or Xaa84 is not the same as the amino acid at the corresponding position in SEQ ID NO;

1.

3. The recombinant antibody of claim 1, wherein said light chain framework region is selected from the group consisting of:

(i) amino acid residues 1-23 of SEQ ID NO;

8; and

(ii) amino acid residues 1-23 of SEQ ID NO;

9.

4. The recombinant antibody of claim 3, wherein said light chain comprises amino acids 1-106 of SEQ ID NO:

9.

8. The recombinant antibody of claim 1 or 2 wherein said antibody has a Kd for EpCAM of at least 10⁻

8 M.

9. The recombinant antibody of claim 1 or 2 wherein said antibody comprises a cytokine.

10. The recombinant antibody of claim 9 wherein said cytokine is IL-2.

11. The recombinant antibody of claim 1 wherein said antibody comprises an amino acid sequence selected from the group consisting of:

(i) amino acid residues 24-31 of SEQ ID NO;

1;

(ii) amino acid residues 49-55 of SEQ ID NO;

1; and

(iii) amino acid residues 88-96 of SEQ ID NO;

1.

13. An expression vector encoding an antibody of claim 1 or 2.

16. A method of treating a human patient having a disease associated with EpCAM over-expression, said method comprising the step of administering an antibody of claim 1 or 2 to a patient.

17. The method of claim 16, wherein said antibody further comprises a cytokine.

18. The method of claim 17, wherein said antibody is administered as an antibody-cytokine fusion protein.

2. A recombinant anti-EpCAM antibody, wherein the antibody comprises an amino acid sequence defining an immunoglobulin heavy chain framework region selected from the group consisting of:

(i) amino acid residues 1-25 of SEQ ID NO;

6, wherein Xaa2 is I or V, Xaa9 is P or A, Xaa11 is L or V, or Xaa17 is T or S, provided that at least one of the amino acid residues at positions Xaa2, Xaa9, Xaa11 or Xaa17 is not the same as the amino acid at the corresponding position in SEQ ID NO;

2;

(ii) amino acid residues 36-49 of SEQ ID NO;

6, wherein Xaa38 is K or R, Xaa40 is T or A, or Xaa46 is K or E, provided that at least one of the amino acid residues at positions Xaa38, Xaa40, Xaa46 is not the same as the amino acid at the corresponding position in SEQ ID NO;

2;

(iii) amino acid residues 67-98 of SEQ ID NO;

6, wherein Xaa68 is F or V, Xaa69 is A or T, Xaa70 is F or I, Xaa73 is E or D, Xaa76 is A or T, Xaa80 is F or Y, Xaa83 is I or L, Xaa84 is N or S, Xaa85 is N or S, Xaa88 is N, A or S, Xaa91 is M or T, or Xaa93 is T or V, provided that at least one of the amino acid residues at positions Xaa68, Xaa69, Xaa70, Xaa73, Xaa76, Xaa80, Xaa83, Xaa84, Xaa85, Xaa88, Xaa91 or Xaa93 is not the same as the amino acid at the corresponding position in SEQ ID NO;

2; and

(iv) amino acid residues 106-116 of SEQ ID NO;

6, wherein Xaa108 is Q or T.

5. The recombinant antibody of claim 2, wherein said heavy chain framework region is selected from the group consisting of:

(i) amino acid residues 1-25 of SEQ ID NO;

18; and

(ii) amino acid residues 67-98 of SEQ ID NO;

18.

6. The recombinant antibody of claim 5, wherein said heavy chain comprises amino acids 1-116 of SEQ ID NO:

18.

12. The recombinant antibody of claim 2 wherein said antibody comprises an amino acid sequence selected from the group consisting of:

(i) amino acid residues 26-35 of SEQ ID NO;

2;

(ii) amino acid residues 50-62 of SEQ ID NO;

2; and

(iii) amino acid residues 101-105 of SEQ ID NO;

2.

7. A recombinant anti-EpCAM antibody comprising light chain amino acid residues 1-106 of SEQ ID NO:

9 and heavy chain amino acid residues of SEQ ID NO;

18.

14. An expression vector encoding the antibody of claim 7.

15. An expression vector having a nucleotide sequence set forth in SEQ ID NO:

32.