Hybrid molecules having translocation region and cell-binding region

• US 5,668,255 A
• Filed: 08/04/1993
• Issued: 09/16/1997
• Est. Priority Date: 06/07/1984
• Status: Expired due to Term

• Current Assignee: Seragen, Inc. (Ligand Pharmaceuticals, Inc.)
• Original Assignee: Seragen, Inc. (Ligand Pharmaceuticals, Inc.)
• Inventors: Murphy, John R.
• Primary Examiner(s): Elliott, George C.
• Assistant Examiner(s): SCHWARTZMAN, ROBERT A
• Application Number:
  US08/102,387
• Time in Patent Office: 1,504 Days
• Field of Search: 424/94.1, 424/85.2, 435/69.1, 435/172.3, 435/320.1, 435/69.7, 530/350, 530/324, 530/351, 530/402, 530/408
• US Class Current: 530/350
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 14/245 Escherichia (G)
• CPC Class Codes: C07K 14/25 Shigella (G)
• CPC Class Codes: C07K 14/28 from Vibrionaceae (F)
• CPC Class Codes: C07K 14/34 from Corynebacterium (G)
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C07K 14/55 IL-2
• CPC Class Codes: C07K 14/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/55 containing a fusion with a toxin, e.g. diphteria toxin
• CPC Class Codes: C07K 2319/75 containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 2740/16022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2740/16222 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2770/24122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12Y 114/16001 Phenylalanine 4-monooxygenase (1.14.16.1)
• CPC Class Codes: C12Y 302/0102 Alpha-glucosidase (3.2.1.20)
• CPC Class Codes: C12Y 302/01052 Beta-N-acetylhexosaminidase (3.2.1.52)
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/68 : involving proteins, peptides or amino acids involving lipoproteins G01N33/92

Abstract

A hybrid molecule including a first part and a second part connected by a covalent bond,

(a) the first part including a portion of the binding domain of a cell-binding ligand, which portion is able to cause the hybrid molecule of the invention to bind to an animal cell; and

(b) the second part including a portion of a translocation domain of a protein, provided that (i) the hybrid molecule does not include an enzymatically-active portion of the protein, (ii) the first part and the second part are not segments of the same naturally-occurring polypeptide toxin, and (iii) the portion of the translocation domain, when covalently bonded to the enzymatically-active effector region of a toxin selected from diphtheria toxin, Pseudomonas exotoxin A, cholera toxin, ricin toxin, and Shiga-like toxin, is capable of translocating such effector region across the cytoplasmic membrane of the cell.

Claims

1. A hybrid molecule comprising a first part and a second part connected via a covalent bond,

(a) said first part comprising a portion of the binding domain of a cell binding ligand effective to cause said hybrid molecule to bind to a cell of an animal;

and

(b) said second part comprising a portion of the translocation domain of a protein, provided that said hybrid molecule does not include an enzymatically active portion of said protein, and wherein said first part and said second part are not segments of the same naturally occurring protein.

2. The hybrid molecule of claim 1, wherein said cell-binding ligand is a polypeptide.

3. The hybrid molecule of claim 1, wherein said protein of (b) is a naturally-occurring toxin.

4. The hybrid molecule of claim 2, wherein said first part comprises the binding domain of said polypeptide cell-binding ligand.

5. The hybrid molecule of claim 2, wherein said first part comprises the cell-binding polypeptide ligand.

6. The hybrid molecule of claim 2, wherein said cell-binding polypeptide binding ligand is a hormone or growth factor.

7. The hybrid molecule of claim 2, wherein said cell-binding polypeptide ligand is an antigen-binding, single-chain analog of a monoclonal antibody.

8. The hybrid molecule of claim 2, wherein said first part comprises a monoclonal antibody.

9. The hybrid molecule of claim 2, wherein said first part comprises a portion of the binding domain of a polypeptide toxin.

10. The hybrid molecule of claim 6, wherein said hormone is selected from the group consisting of insulin, interleukin II, interleukin IV, interleukin VI and EGF.

11. The hybrid molecule of claim 10, wherein said hormone is interleukin II.

12. The hybrid molecule of claim 10, wherein said hormone is EGF.

13. The hybrid molecule of claim 9, wherein said toxin is diphtheria toxin.

14. The hybrid molecule of claim 3, wherein said toxin is diphtheria toxin.

15. The hybrid molecule of claim 3, wherein said toxin is botulinum neurotoxin.

16. The hybrid molecule of claim 3, wherein said toxin is cholera toxin.

17. The hybrid molecule of claim 3, wherein said toxin is ricin toxin.

18. The hybrid molecule of claim 3, wherein said toxin is Shiga toxin.

19. The hybrid molecule of claim 3, wherein said toxin is Shiga-like toxin.

20. The hybrid molecule of claim 3, wherein said toxin is Pseudomonas exotoxin A.