Mutant BR96 antibodies reactive with human carcinomas

• US 5,792,456 A
• Filed: 06/07/1995
• Issued: 08/11/1998
• Est. Priority Date: 08/04/1994
• Status: Expired due to Fees

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: Glaser, Scott, Huse, William, Rosok, Mae Joanne, Yelton, Dale
• Primary Examiner(s): Feisee, Lila
• Assistant Examiner(s): UNGAR, SUSAN NMN
• Application Number:
  US08/487,860
• Time in Patent Office: 1,161 Days
• Field of Search: 530/388.1, 530/387.1, 530/387.3, 530/388.8, 530/391.3, 530/391.7, 424/130.1, 424/133.1, 424/141.1
• US Class Current: 424/133.1
• 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: A61K 51/1045 against animal or human tumor cells or tumor cell determinants
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

The present invention provides mutant BR96 polypeptides having a variable region comprising an amino acid sequence derived from the variable region of BR96.

Claims

1. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR3 is mutated to have therein the sequence represented by Gly Leu Xaa Asp Gly Ala Trp (SEQ ID NO;

49) wherein Xaa is an amino acid selected from the group consisting of alanine, arginine, serine, glycine, tyrosine, valine, beginning at amino acid Kabat position 95 and ending with amino acid position Kabat 100a as shown in Table 1.

2. The mutant polypeptide of claim 1 further comprising a CDR including a sequence Gly Phe Pro Phe Ala Ser Tyr Tyr (SEQ ID NO:

50) beginning at amino acid position 26 and ending with amino acid position 33 as shown in FIG. 4.

3. The mutant polypeptide of claim 1 further comprising a CDR including a sequence Ser Gln Asp Gly Asp Ile Thr Asp (SEQ ID NO:

60) beginning at amino acid position 52 and ending with amino acid position 59 as shown in FIG. 5.

4. The mutant polypeptide of claim 1 further comprising a CDR including a sequence Gly Phe Pro Phe Ser Asp Tyr Tyr (SEQ ID NO:

59) beginning at amino acid position 26 and ending with amino acid position 33 as shown in FIG. 25 and a CDR including a sequence Ser Gln Asp Gly Asp Ile Thr Asp (SEQ ID NO;

     60) beginning at amino acid position 52 and ending with amino acid position 59 as shown in FIG. 25.

20. A substantially purified polypeptide of claim 1, 5, or 16.

21. A mutant polypeptide antibody molecule of claim 1, 5, or 16.

22. A monoclonal antibody of claim 21.

23. A chimeric monoclonal antibody of claim 22.

24. A mutant Fab polypeptide of claim 1, 5, or 16.

25. A mutant F(ab'"'"')₂ polypeptide comprising the polypeptide of claim 1, 5, or 16.

26. A mutant Fv polypeptide of claim 1, 5, or 16.

27. An immunoconjugate comprising the polypeptide of claim 1, 5, or 16 joined to a cytotoxic agent.

28. A composition comprising the immunoconjugate of claim 27 in a pharmaceutically acceptable carrier.

29. The immunoconjugate of claim 27, wherein the cytotoxic agent is selected from the group consisting of antimetabolites, alkylating agents, anthracyclines, antibiotics, anti-mitotic agents, and chemotherapeutic agents.

30. The immunoconjugate of claim 27, wherein the cytotoxic agent is selected from the group consisting of ricin, doxorubicin, daunorubicin, taxol, ethidium bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, dihydroxy anthracin dione, actinomycin D, 1-dehydrotestosterone, and glucocorticoid.

5. A BR96 (ATCC:

HB10036) polypeptide comprising a heavy chain variable region wherein the heavy chain variable region is mutated to have therein the sequence represented by Gly Leu Xaa Asp Gly Ala Trp (SEQ ID NO;

49) wherein Xaa is an amino acid selected from the group consisting of alanine, arginine, serine, glycine, tyrosine, valine, beginning at amino acid Kabat position 95 and ending with amino acid Kabat position 10a as shown in Table 1.

6. The mutant polypeptide of claim 5 further comprising a heavy chain variable region including therein the sequence Gly Phe Pro Phe Ala Ser Tyr Tyr (SEQ ID NO:

50) beginning at amino acid position 26 and ending with amino acid position 33 as shown in FIG. 4.

7. The mutant polypeptide of claim 5 further comprising a heavy chain variable region including therein the sequence Ser Gln Asp Gly Asp Ile Thr Asp (SEQ ID NO:

60) beginning at amino acid position 52 and ending with amino acid position 59 as shown in FIG. 5.

8. The mutant polypeptide of claim 5 further comprising a heavy chain variable region including therein a sequence Gly Phe Pro Phe Ser Asp Tyr Tyr (SEQ ID NO:

59) beginning at amino acid position 26 and ending with amino acid position 33 as shown in Table 1 and a sequence Ser Gln Asp Gly Asp Ile Thr Asp (SEQ ID NO;

     60) beginning at amino acid position 52 and ending with amino acid position 59 as shown in FIG. 25.

9. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR1 includes therein the sequence represented by Gly Phe Thr Phe Ser Asp Tyr Tyr Met Tyr (SEQ ID NO;

     51), wherein CDR2 is mutated to have therein the sequence represented by Tyr Ile Ser Gln Xaa Gly Asp Ile Thr Asp Tyr Pro Asp Thr Val Lys Gly (SEQ ID NO;

     52), wherein Xaa is selected from the group consisting of glycine or aspartic acid, and wherein CDR3 is mutated to have therein the sequence represented by Gly Leu Ala Asp Gly Ala Trp (SEQ ID NO;

     53) as shown in FIG. 2.

10. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR1 includes therein the sequence represented by Gly Phe Thr Phe Ser Asp Tyr Tyr Met Tyr (SEQ ID NO;

     51), and wherein CDR3 is mutated to have therein the sequence represented by Gly Leu Ala Asp Gly Ala Trp (SEQ ID NO;

     53) as shown in FIG. 2.

11. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR2 is mutated to have therein the sequence represented by Tyr Ile Ser Gln Xaa Gly Asp Ile Thr Asp Tyr Pro Asp Thr Val Lys Gly (SEQ ID NO;

     52) wherein Xaa is selected from the group consisting of glycine or aspartic acid and wherein CDR3 is mutated to have therein the sequence represented by Gly Leu Ala Asp Gly Ala Trp (SEQ ID NO;

     53) as shown in FIG. 2.

12. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR2 is mutated to have therein the sequence represented by Tyr Ile Ser Gln Asp Gly Asp Ile Thr Asp Tyr Pro Asp Thr Val Lys Gly (SEQ ID NO;

     54) beginning at amino acid position 50 and ending with amino acid position 66 as shown in FIG. 3.

13. A BR96 (ATCC:

HB10036) polypeptide comprising a heavy chain variable region wherein the heavy chain variable region is mutated to have therein the sequence represented by Tyr Ile Ser Gln Asp Gly Asp Ile Thr Asp Tyr Pro Asp Thr Val Lys Gly (SEQ ID NO;

     54) beginning at amino acid position 50 and ending with amino acid position 66 as shown in FIG. 3.

14. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR1 includes therein the sequence represented by Gly Phe Thr Phe Ser Asp Tyr Tyr Met Tyr (SEQ ID NO;

     51), wherein CDR2 is mutated to have therein the sequence represented by Tyr Ile Ser Gln Asp Gly Asp Ile Thr Asp Tyr Pro Asp Thr Val Lys Gly (SEQ ID NO;

     54), and wherein CDR3 includes therein the sequence represented by Gly Leu Asp Asp Gly Ala Trp (SEQ ID NO;

     55) as shown in FIG. 3.

15. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR2 is mutated to have therein the sequence represented by Tyr Ile Ser Gln Asp Gly Asp Ile Thr Asp Tyr Pro Asp Thr Val Lys Gly (SEQ ID NO;

     54) and wherein CDR3 includes therein the sequence represented by Gly Leu Asp Asp Gly Ala Trp (SEQ ID NO;

     55) as shown in FIG. 3.

16. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR1 is mutated to have therein the sequence represented by Gly Phe Pro Phe Ser Asp Tyr Tyr (SEQ ID NO;

     59), wherein CDR2 is mutated to have therein the sequence represented by Ser Gln Asp Gly Asp Ile Thr Asp (SEQ ID NO;

     60), and wherein CDR3 is mutated to have therein the sequence represented by Gly Leu Ala Asp Gly Ala Trp (SEQ ID NO;

     53) as shown in FIG. 25.

17. A BR96 (ATCC HB10036) polypeptide comprising CDR1, CDR2, and CDR3, wherein CDR1 includes therein the sequence represented by Gly Phe Thr Phe Ser Asp Tyr Tyr (SEQ ID NO:

51), wherein CDR2 is mutated to have therein the sequence represented by Ser Gln Asp Gly Asp Ile Thr Asp (SEQ ID NO;

     60), and wherein CDR3 is mutated to have therein the sequence represented by Gly Leu Ala Asp Gly Ala Trp (SEQ ID NO;

     53) as shown in FIG. 5.

18. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR1 is mutated to have therein the sequence represented by Gly Phe Pro Phe Ala Ser Tyr Tyr (SEQ ID NO;

     50), wherein CDR2 includes therein the sequence represented by Ser Gln Gly Gly Asp Ile Thr Asp (SEQ ID NO;

     62), and wherein CDR3 includes therein the sequence represented by Gly Leu Asp Asp Gly Ala Trp (SEQ ID NO;

     55) as shown in Table 1.

19. A BR96 (ATCC:

HB10036) polypeptide comprising CDR1, CDR2 and CDR3, wherein CDR1 is mutated to have therein the sequence represented by Gly Phe Pro Phe Ala Ser Tyr Tyr (SEQ ID NO;

     50), wherein CDR2 includes therein the sequence represented by Ser Gln Gly Gly Asp Ile Thr Asp (SEQ ID NO;

     62), and wherein CDR3 is mutated to have therein the sequence represented by Gly Leu Ala Asp Gly Ala Trp (SEQ ID NO;

     53) as shown in FIG. 4.