Branched peptide linkers

• US 6,759,509 B1
• Filed: 10/31/1997
• Issued: 07/06/2004
• Est. Priority Date: 11/05/1996
• Status: Expired due to Fees

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: King, Dalton, Dubowchik, Gene M., Firestone, Raymond A.
• Primary Examiner(s): Low, Christopher S. F.
• Assistant Examiner(s): Lukton, David
• Application Number:
  US08/962,348
• Time in Patent Office: 2,440 Days
• Field of Search: 514/18, 514/19, 514/2, 530/330, 530/331, 530/399, 530/345, 424/178.1, 424/179.1, 424/181.1
• US Class Current: 530/330
• 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 47/641 Branched, dendritic or hypercomb peptides
• CPC Class Codes: A61K 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: C07K 5/0202 containing the structure -NH-X-X-C(=0)-, X being an optionally substituted carbon atom or a heteroatom, e.g. beta-amino acids
• CPC Class Codes: C07K 5/06026 the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
• CPC Class Codes: C07K 5/06052 Val-amino acid
• CPC Class Codes: C07K 5/06078 and aromatic or cycloaliphatic
• CPC Class Codes: C07K 5/06086 with the first amino acid being basic

Abstract

Conjugates containing a targeting ligand, such as an antibody, a therapeutically active drug and a branched peptide linker. The branched peptide linker contains two or more amino acid moieties that provide an enzyme cleavage site. The number of drugs capable of being bonded to the branched linkers varies by a factor of two for each generation of branching.

Claims

1. A compound of the following formula II:

wherein

A is a thiol acceptor;

W is a bridging moiety;

c is an integer of 0 to 1;

a is an integer of 2 to 12;

Q is O, NH, or N-lower alkyl;

p is an integer of 0 or 1;

d is an integer of 0 or 1;

E is a polyvalent atom;

each b is an integer of 1 to 10;

each X is of the formula

2. The compound of claim 1 wherein each X is of the formula

3. The compound of claim 1 wherein each X¹ is of the formula

4. The compound of claim 1 wherein each X² is of the formula

5. The compound of claim 1 wherein each X³ is of the formula

6. The compound of claim 1 wherein —

Y—

Z_m is a dipeptide, tripeptide or tetrapeptide containing amino acids selected from the group consisting of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, lysine, lysine protected with acetyl or formyl, arginine, arginine protected with tosyl or nitro groups, histidine, ornithine, ornithine protected with acetyl or formyl, and citrulline.

7. The compound of claim 1 wherein the —

Y—

Z_m group is phenylalanine-lysine.

9. The compound of claim 1 wherein the —

Y—

Z_m group is valine-lysine.

10. The compound of claim 1 wherein E is nitrogen.

11. The compound of claim 1 wherein E is carbon.

12. The compound of claim 1 wherein D is an amino group-containing drug moiety selected from the group consisting of mitomycin-C, mitomycin-A, daunorubicin, doxorubicin, N-(5,5-diacetoxypentyl)doxorubicin, aminopterin, actinomycin, bleomycin, 9-amino camptothecin, N8-acetyl spermidine, 1-(2chloroethyl)-1,2-dimethanesulfonyl hydrazide, tallysomycin, and derivatives thereof.

13. The compound of claim 1 wherein D is doxorubicin.

14. The compound of claim 1 wherein D is mitomycin-C.

15. The compound of claim 1 wherein D is mitomycin-A.

16. The compound of claim 1 wherein D is tallysomycin.

17. The compound of claim 1 wherein D is N-(5,5-diacetoxypentyl)doxorubicin.

18. The compound of claim 1 wherein D is a hydroxyl containing drug moiety selected from the group consisting of etoposide, camptothecin, taxol, esperamicin, 1,8-dihydroxy-bicyclo[7.3.1]trideca-4-ene-2,6-diyne-13-one, anguidine, doxorubicin, morpholine-doxorubicin, N-(5,5-diacetoxypentyl)doxorubicin, vincristine, vinblastine and derivatives thereof.

19. The compound of claim 1 wherein D is a sulfhydryl containing drug moiety selected from the group consisting of esperamicin and 6-mercaptopurine, and derivatives thereof.

20. The compound of claim 1 wherein D is a carboxyl containing drug moiety selected from the group consisting of methotrexate, campothecin (ring-opened form of the lactone), butyric acid, retinoic acid, and derivatives thereof.

21. The compound of claim 1 wherein D is an anthracycline.

22. The compound of claim 1 wherein G is

23. The compound of claim 1 wherein G is p-aminobenzyl-carbonyl.

24. The compound of claim 1 wherein G has the formula

25. The compound of claim 1 wherein G has the formula

wherein

T is O, N or S;

and

R² is H or C₁-C₅ alkyl.

8. The compound of claimn 1 wherein the —

Y_m—

Z_m group is valine-citrulline.

26. A compound of the following formula III:

wherein

L is a ligand;

q is an integer of 1 to 10;

A is a thiol acceptor;

W is a bridging moiety;

c is an integer from 0 to 1;

a is an integer of 2 to 12;

Q is O, NH, or N-lower alkyl;

p is an integer of 0 or 1;

d is an integer of 1 or 2;

E is a polyvalent atom;

each b is an integer of 1 to 10;

each X is of the formula

27. The compound of claim 26 wherein L is an immunoglobulin, or antigen-recognizing fragment thereof.

28. The branched peptide linker of claim 27 covalently bonded to an antibody or antigen-recognizing fragment thereof.

29. The branched peptide linker of claim 26 constantly bonded to drug moieties which are anthracyclines.

30. The compound of claim 26 wherein L is an immunoglobulin selected from the group consisting of BR96, BR64, L6, a relaxed BR96, a relaxed BR64, a chimeric L6, a relaxed chimeric BR96, a relaxed chimeric BR64, a relaxed chimeric L6;

and antigen-recognizing fragments thereof.

31. The compound of claim 26 wherein L is a ligand selected from the group consisting of bombesin, EDG, transferrin, gastrin, gastrin-releasing peptide, platelet-derived growth factor, IL-2, II-6, TFG-α

, TFG-β

, VGF, insulin, insulin-like growth factors I and II, carbohydrates, lectins, and apoproteins from low density lipoproteins.

32. A pharmaceutical composition comprising a compound of claim 26 and a pharmaceutically acceptable carrier.

33. A method of controlling the growth of cells which comprises administering a compound of claim 26 to a host in need of treatment.

34. The method of claim 33 wherein the cells are cancer cells.

35. A method of cancer treatment of a warm blooded animal in need thereof which comprises administering to said animal an effective tumor inhibiting amount of a compound of claim 26.