Prodrugs activated by plasmin and their use in cancer chemotherapy

• US 7,893,023 B2
• Filed: 06/11/2008
• Issued: 02/22/2011
• Est. Priority Date: 08/24/2000
• Status: Active Grant

• Current Assignee: ER Squibb & Sons LLC (Bristol-Myers Squibb Company)
• Original Assignee: Medarex Incorporated (Bristol-Myers Squibb Company)
• Inventors: Dubois, Vincent, Trouet, Andre, Passioukov, Alexandre
• Primary Examiner(s): Canella; Karen A
• Application Number:
  US12/157,575
• Publication Number:
  US 20090076176A1
• Time in Patent Office: 986 Days
• Field of Search: None
• US Class Current: 514/1.3
• CPC Class Codes: A61K 38/06 Tripeptides
• CPC Class Codes: A61K 38/07 Tetrapeptides
• CPC Class Codes: A61K 38/08 Peptides having 5 to 11 amino acids A61K38/043 - A61K38/046 take precedence
• CPC Class Codes: A61K 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: A61K 47/67 Enzyme prodrug therapy, e.g. gene directed enzyme drug therapy [GDEPT] or VDEPT
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Abstract

The product of the invention is a modified form of a therapeutic agent and comprises a therapeutic agent, an oligopeptide having a plasmin peptide substrate of 2-4 amino acids and mono- or di-peptide linkage, a stabilizing group and, optionally, a linker group. The prodrug is cleavable by plasmin. Also disclosed are methods of making and using the prodrug compounds.

Claims

1. A method for treating a patient having a medical condition associated with plasmin overproduction, the method comprising administering to the patient a compound comprising:

(1) a therapeutic agent capable of entering a target cell;

(2) an oligopeptide having the formula (AA^x)_m-(AA^y)_n (SEQ ID NO;

     1) wherein;

(AA^x)_m is a plasmin substrate and each AA^x independently represents an amino acid selected from the group consisting of Leu, Lys, Val, Phe, Ala, Glu, Be, Gly, Pro and Arg;

each AA^y independently represents an amino acid selected from the group consisting of Ile, Leu, Phe and Val;

m is an integer from 2-4; and

n is an integer from 1-2;

(3) a stabilizing group; and

(4) optionally, a linker group not cleavable by plasmin;

wherein the oligopeptide is directly linked to the stabilizing group at a first attachment site of the oligopeptide and the oligopeptide is directly linked to the therapeutic agent or indirectly linked through the linker group to the therapeutic agent at a second attachment site of the oligopeptide;

wherein the stabilizing group hinders cleavage of the oligopeptide by enzymes present in whole blood; and

wherein the compound is cleaved by plasmin under physiological conditions.

2. The method of claim 1 wherein the compound is administered intravenously.

3. The method of claim 1 wherein the patient is treated for a medical condition selected from the group consisting of cancer, neoplastic diseases, tumors, inflammatory diseases, and infectious diseases.

4. A method for decreasing the toxicity of a therapeutic agent wherein the therapeutic agent is intended for administration to a patient having a medical condition associated with plasmin overproduction, the method comprising:

covalently forming a prodrug by linking an oligopeptide cleavable by plasmin to a stabilizing group at a first attachment site of the oligopeptide and directly or indirectly linking the therapeutic agent at a second attachment site of the oligopeptide, wherein the oligopeptide has the formula (AA^x)_m-(AA^y)_n (SEQ ID NO;

     1) wherein;

(AA^x)_m is a plasmin substrate and each AA^x independently represents an amino acid selected from the group consisting of Leu, Lys, Val, Phe, Ala, Glu, Ile, Gly, Pro and Arg;

each AA^y independently represents an amino acid selected from the group consisting of Ile, Leu, Phe and Val;

m is an integer from 2-4; and

n is an integer from 1-2, and wherein the compound is cleavable by plasmin, whereby the prodrug provides for decreased toxicity of the therapeutic agent when administered to the patient.

5. A compound comprising:

(1) a therapeutic agent capable of entering a target cell;

(2) an oligopeptide having the formula X-Y, where X is a plasmin peptide substrate of 2-4 amino acids independently selected from the group consisting of Leu, Lys, Val, Phe, Ala, Glu, Ile, Gly, Pro and Arg, and Y is a peptide comprising 1-2 amino acids independently selected from the group consisting of Ile, Leu, Phe and Val;

(3) a stabilizing group; and

(4) optionally, a linker group not cleavable by plasmin;

wherein the oligopeptide is directly linked to the stabilizing group at a first attachment site of the oligopeptide and the oligopeptide is directly linked to the therapeutic agent or indirectly linked through the linker group to the therapeutic agent at a second attachment site of the oligopeptide;

wherein the stabilizing group hinders cleavage of the oligopeptide by enzymes present in whole blood; and

wherein the compound is cleavable by plasmin.

6. The compound of claim 5 wherein the target cell is a tumor or inflammatory cell.

7. The compound of claim 5 wherein plasmin is present in the extracellular vicinity of the target cell for the therapeutic agent.

8. The compound of claim 5 wherein plasmin cleaves the linkage between X and Y of the oligopeptide.

9. The compound of claim 5 which is selectively cleavable by plasmin.

10. The compound of claim 5 being a prodrug having an active portion, wherein the active portion of the prodrug is more capable of entering the target cell after cleavage by the plasmin than prior to cleavage by plasmin, the active portion including at least the therapeutic agent.

11. The compound of claim 10 wherein the active portion of the prodrug consists of the therapeutic agent.

12. The compound of claim 10 wherein the active portion of the prodrug includes the therapeutic agent and at least the linker group.

13. The compound of claim 10 wherein the active portion of the prodrug includes the therapeutic agent and one amino acid of the Y portion of the oligopeptide.

14. The compound of claim 13 further comprising two amino acids of the Y portion of the oligopeptide.

15. The compound of claim 5 wherein the oligopeptide is selected from:

Leu-Lys-Leu- and Leu-Lys-Leu-Leu (SEQ ID NO;

     4).

16. The compound of claim 5 wherein X is (AA^x)_m and Y is (AA^y)_n wherein:

(AA^x)_m is a plasmin substrate and each AA^x independently represents an amino acid selected from the group consisting of Leu, Lys, Val, Phe, Ala, Glu, Ile, Gly, Pro and Arg;

each AA^y independently represents an amino acid selected from the group consisting of Ile, Leu, Phe and Val;

m is an integer from 2-4; and

n is an integer from 1-2.

17. The compound of claim 16 wherein AA^x4 is an amino acid residue selected from the group consisting of Ile and Phe.

18. The compound of claim 16 wherein AA^x3 is an amino acid residue selected from the group consisting of Ala, Glu, Gly, Ile, Leu, Phe, Pro and Val.

19. The compound of claim 16 wherein AA^x2 is an amino acid residue selected from the group consisting of Ala, Glu, Gly, Leu, Lys, Phe, Pro and Val.

20. The compound of claim 16 wherein AA^x1 is an amino acid residue selected from the group consisting of Arg and Lys.

21. The compound of claim 16 wherein AA^y2 and AA^y1 are amino acid residues selected from the group consisting of Ile, Leu, Phe and Val.

22. The compound of claim 16 wherein AA^y2 and AA^y1 are Leu residues.

23. The compound of claim 16 wherein AA^y1 is a Leu residue and AA^y2 is absent.

24. The compound of claim 5 wherein the stabilizing group is selected from the group consisting of D-Ala, β

-Ala, α

-methyl-Ala, Succ-D-Ala, Succ-β

-Ala and Succ-α

-methyl-Ala.

25. The compound of claim 5 wherein the therapeutic agent is selected from the group consisting of alkylating agents, anthracyclines, camptothecins, cyclosporins, dolastatins, enediynes, epipodophyllotoxins, maytansinoids, naphtalimides, pteridines, rhodamines, sulfoximines, taxanes, taxoids, and vinca alkaloids.

26. The compound of claim 5 wherein the therapeutic agent is selected from the group consisting of actinomycin D;

alkylating agents;

amiodarone;

anthracyclines;

AT-125 or activin ((α

S,5S)-α

-amino-3-chloro-4,5-dihydro-5-isoxazole acetic acid);

bamipine;

bleomycin;

5-bromodeoxyuridine;

calicheamicin;

camptothecins;

L-canavanine;

carboplatin;

CC-1065;

chlorphenoxamine;

chloroquine;

colchicine;

combretastatin and combretastatin A₄ phosphate;

coumarins;

cyclophosphamide;

cyclosporins;

cytarabine;

dehydrodidemnin B;

dipyridamole;

discodermolide;

docetaxel;

dolastatin 10, dolastatin 11 and dolastatin 15;

duocarmycin;

epothilone A;

etoposide and etoposide phosphate;

fludarabine;

5-fluorouracil;

folic acid derivatives;

KW-2189;

6-maytansinoids;

mitomycin C;

naphtalimides;

nitrosoureas;

paclitaxel;

phenylenediamine mustards;

cis-platin;

podophyllotoxin and podophyllotoxin derivatives;

porfiromycin;

quinidine;

quinine;

reserpine;

rhodamines;

sulfoximines;

tamoxifen;

taxoids;

topotecan;

trifluoperazine;

verapamil;

vinca alkaloids; and

derivatives and analogs thereof.

27. The compound of claim 5 wherein the therapeutic agent is daunorubicin.

28. The compound of claim 5 wherein the therapeutic agent is doxorubicin.

29. The compound of claim 5 wherein the therapeutic agent has an intracellular active site.

30. The compound of claim 5 wherein the oligopeptide is directly linked to the therapeutic agent.

31. The compound of claim 5 wherein the oligopeptide is indirectly linked to the therapeutic agent at the second attachment site of the oligopeptide via a linker group, and the linker group is selected from the group consisting of amino caproic acid, hydrazide group, an ester group, an ether group, and a sulphydryl group.

32. The compound of claim 5 wherein X is a plasmin peptide substrate of 2-4 amino acids independently selected from Leu-Lys, Val-Leu-Lys, Phe-Lys, Ala-Phe-Lys, Ala-Lys, Ala-Ala-Lys, Leu-Lys-Lys, Glu-Lys-Lys, Phe-Glu-Lys-Lys (SEQ ID NO:

2), Glu-Lys, Phe-Glu-Lys, Ile-Glu-Lys, Gly-Gly-Arg, Val-Gly-Arg, Ile-Glu-Gly-Arg (SEQ ID NO;

     3), Pro-Arg, Gly-Pro-Arg, Phe-Val-Arg, Leu-Arg, Phe-Arg and Pro-Phe-Arg.

33. An oligopeptide having the formula (AA^x)_m-(AA^y)_n (SEQ ID NO:

1) wherein;

(AA^x)_m is a plasmin substrate and each AA^x independently represents an amino acid selected from the group consisting of Leu, Lys, Val, Phe, Ala, Glu, Ile, Gly, Pro and Arg;

each AA^x independently represents an amino acid selected from the group consisting of Ile, Leu, Phe and Val;

m is an integer from 2-4; and

n is an integer from 1-2, said oligopeptide being cleavable by plasmin, wherein the oligopeptide is linked to a therapeutic agent.

34. The oligopeptide of claim 33 wherein the oligopeptide is linked to a stabilizing group.

35. An oligopeptide having the formula (AA^x)_m-(AA^y)_n (SEQ ID NO:

1) wherein;

(AA^x)_m is plasmin substrate and each AA^x independently represents an amino acid selected from the group consisting of Leu, Lys, Val, Phe, Ala, Glu, Ile, Gly, Pro and Arg;

each AA^y independently represents an amino acid selected from the group consisting of Ile, Leu, Phe and Val;

m is an integer from 2-4; and

n is an integer from 1-2, said oligopeptide being cleavable by plasmin, wherein the oligopeptide is linked to a P-glycoprotein inhibitor.

36. An oligopeptide having the formula (AA^x)_m-(AA^y)_n (SEQ ID NO:

1) wherein;

(AA^x)_m is a plasmin substrate and each AA^x independently represents an amino acid selected from the group consisting of Leu, Lys, Val, Phe, Ala, Glu, Ile, Gly, Pro and Arg;

each AA^y independently represents an amino acid selected from the group consisting of Ile, Leu, Phe and Val;

m is an integer from 2-4; and

n is an integer from 1-2, said oligopeptide being cleavable by plasmin, wherein the oligopeptide is linked to a γ

-glutamylcysteine synthetase inhibitor.

37. A pharmaceutical composition comprising:

(1) A compound comprising;

(a) a therapeutic agent capable of entering a target cell;

(b) an oligopeptide having the formula (AA^x)_m-(AA^y)_n (SEQ ID NO;

     1) wherein;

(AA^x)_m is a plasmin substrate and each AA^x independently represents an amino acid selected from the group consisting of Leu, Lys, Val, Phe, Ala, Glu, Ile, Gly, Pro and Arg;

each AA^y independently represents an amino acid selected from the group consisting of Ile, Leu, Phe and Val;

m is an integer from 2-4; and

n is an integer from 1-2;

(c) a stabilizing group; and

(d) optionally, a linker group not cleavable by plasmin;

wherein the oligopeptide is directly linked to the stabilizing group at a first attachment site of the oligopeptide and the oligopeptide is directly linked to the therapeutic agent or indirectly linked through the linker group to the therapeutic agent at a second attachment site of the oligopeptide;

wherein the stabilizing group hinders cleavage of the oligopeptide by enzymes present in whole blood; and

wherein the compound is cleaved by plasmin under physiological conditions; and

(2) a pharmaceutically acceptable carrier.