Prodrug compounds with an isoleucine

• US 7,115,573 B2
• Filed: 06/11/2001
• Issued: 10/03/2006
• Est. Priority Date: 06/14/2000
• Status: Expired due to Term

• Current Assignee: ER Squibb & Sons LLC (Bristol-Myers Squibb Company)
• Original Assignee: Medarex Incorporated (Bristol-Myers Squibb Company)
• Inventors: Gangwar, Sanjeev, Lobl, Thomas J., Yarranton, Geoffrey T., Pickford, Lesley B., Nieder, Matthew H.
• Primary Examiner(s): Russel, Jeffrey Edwin
• Application Number:
  US10/311,411
• Publication Number:
  US 20040039160A1
• Time in Patent Office: 1,940 Days
• Field of Search: 530/329, 530/330, 530/331, 530/345, 514/17, 514/18
• US Class Current: 514/1.3
• CPC Class Codes: A61K 38/06 Tripeptides
• CPC Class Codes: A61K 38/07 Tetrapeptides
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61K 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 5/0808 the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
• CPC Class Codes: C07K 5/101 the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
• CPC Class Codes: C07K 5/1024 with the first amino acid being heterocyclic

Abstract

The compounds of the invention are modified forms of therapeutic agents. A typical prodrug compound of the invention comprises a therapeutic agent, an oligopeptide having an isoleucine residue, a stabilizing group and, optionally, a linker group. The prodrug is cleavable by an enzyme associated with the target cell. Methods of making and using the compounds are also disclosed.

Claims

1. A compound comprising:

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

(2) an oligopeptide of the formula (AA)_n-AA³-AA²-AA¹, wherein;

AA¹ is selected from the group consisting of Leucine, Phenylalanine, Isoleucine, Alanine, Glycine, Tyrosine, 2-Naphthylalanine, Serine, Proline, and β

-Alanine,

AA² represents any amino acid,

AA³ represents isoleucine,

n is 0 or 1, and when n is 1 then (AA)_n is AA⁴ which represents any amino acid,

(3) a negatively charged stabilizing group, and

(4) optionally, a linker group not cleavable by TOP, 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 compound by enzymes present in whole blood, and

wherein the compound is cleavable by an enzyme associated with the target cell, the enzyme associated with the target cell being other than TOP,

or a salt of said compound.

3. The compound of claim 1 or 2, wherein the compound is resistant to cleavage by TOP.

4. The compound of claim 1 wherein AA² of the oligopeptide is selected from Alanine, Leucine, Glycine, Serine, Tyrosine, 3-Pyridylalanine, 2-Thienylalanine, N-Methylalanine, Aminoisobutyric Acid, Threonine, and Phenylalanine.

5. The compound of claim 1 or 2, wherein AA⁴ is selected from the group consisting of β

-Alanine, Thiazolidine-4-carboxylic acid, 2-Thienylalanine, 2-Naphthylalanine, D-Alanine, D-Leucine, D-Methionine, D-Phenylalanine, 3-Amino-3-phenylpropionic acid, γ

-Aminobutyric acid, 3-amino-4,4-diphenylbutyric acid, Tetrahydroisoquinoline-3-carboxylic acid, 4-Aininomethylbenzoic acid, and Aminoisobutyric acid.

6. The compound of claim 1 or 2, wherein the oligopeptide is β

Ala-Ile-Ala-Leu (SEQ ID NO;

8).

7. The compound of claim 1 or 2, wherein the stabilizing group is a dicarboxylic or higher order carboxylic acid.

8. The compound of claim 1, wherein the stabilizing group is selected from the group consisting of succinic acid, adipic acid, glutaric acid, phthalic acid, diglycolic acid, fumaric acid, naphthalene dicarboxylic acid, 1,8-naphthyl dicarboxylic acid, aconitic acid, carboxycinnamic acid, triazole dicarboxylic acid, 4-carboxyphenyl boronic acid, butane disulfonic acid, and maleic acid.

9. The compound of claim 1 or 2, wherein the stabilizing group is one of aspartic acid linked to the oligopeptide at the β

-carboxy group of the aspartic acid or glutamic acid linked to the oligopeptide at the γ

-carboxy group of the glutamic acid.

10. The compound of claim 1 or 2, wherein the enzyme associated with the target cell cleaves the linkage between AA¹ and AA² of the oligopeptide.

11. The compound of claim 1 or 2, wherein the oligopeptide is directly linked to the therapeutic agent.

12. The compound of claim 1 or 2, wherein the oligopeptide sequence is indirectly linked to the therapeutic agent at the second attachment site of the oligopeptide via a linker group, the linker group selected from amino caproic acid, hydrazide group, an ester group, an ether group, or a suifflydryl group.

13. The compound of claim 1 or 2, wherein the therapeutic agent is selected from the group consisting of Alkylating Agents, Antiproliferative agents, Tubulin Binding agents, Vinca Alkaloids, Enedjynes, Podophyllotoxins or Podophyllotoxin derivatives, the Pteridine family of drugs, Taxanes, Anthracyclines, Dolastatins, Topoiosomerase inhibitors, and Platinum coordination-complex chemotherapeutic agents.

14. The compound of claim 1 or 2, wherein the therapeutic agent is selected from Doxorubicin, Daunorubicin, Vinblastine, Vincristine, Calicheamicin, Etoposide, Etoposide phosphate, CC-1065, Duocarmycin, KW-2189, Methotrexate, Methopterin, Aminopterin, Dichloromethotrexate, Docetaxel, Paclitaxel, Epithiolone, Combretastatin, Combretastatin A₄ Phosphate, Dolastatin 10, Dolastatin 11, Dolastatin 15, Topotecan, Camptothecin, Mitomycin C, Porfiromycin, 5-Fluorouracil, 6-Mercaptopurine, Fludarabine, Tamoxifen, Cytosine arabinoside, Adenosine arabinoside, Colchicine, Cisplatin, Carboplatin, Bleomycin, Melphalan, Chloroquine, Cyclosporin A, a derivative of any of the foregoing and an analog of any of the foregoing.

15. The compound of claim 1 or 2, wherein the therapeutic agent has an intracellular active site.

16. The compound of claim 1 or 2, wherein the compound is selected from the group consisting of:

Suc-β

Ala-Ile-Ala-Leu-Dnr (SEQ ID NO;

     43), Suc-β

Ala-Ile-Ala-Leu-Dox (SEQ ID NO;

     39), Suc-Ile-Ala-Leu-Dox, Suc-Ile-N(Me)Ala-Leu-Dox, and Suc-Ile-Ala-Gly-Dox.

17. The compound of claim 1 or 2, wherein the target cell is a tumor cell or inflammatory cell.

18. The compound of claim 1 or 2, wherein the enzyme associated with the target cell is present in the extracellular vicinity of the target cell for the therapeutic agent.

2. A compound comprising:

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

(2) an oligopeptide of the formula (AA)_n-AA³-AA²-AA¹, wherein;

AA¹ is selected from the group consisting of Leucine, Phenylalanine, Isoleucine, Alanine, Glycine, Tyrosine, 2-Naphthylalanine, Serine, Proline, and β

-Alanine,

AA² is selected from group consisting Alanine, Leucine, Glycine, Serine, 3-Pyndylalanine, 2-Thienylalanine, Tyrosine, N-Methylalanine, Aminoisobutyric Acid, Threonine, and Phenylalanine,

AA³ represents isoleucine, and

n is 0 or 1, and when n is 1, then (AA)_n is AA⁴ which represents any amino acid,

(3) a neutral stabilizing group or a negatively charged stabilizing group, and

(4) optionally, a linker group not cleavable by TOP,

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 compound by enzymes present in whole blood, and

wherein the compound is cleavable by an enzyme associated with the target cell, the enzyme associated with the target cell being other than TOP, or a salt of said compound.

19. The compound of claim 2 wherein the stabilizing group is negatively charged.

20. The compound of claim 2, wherein the stabilizing group is selected from the group consisting of succinic acid, adipic acid, glutaric acid, phthalic acid, diglycolic acid, fumaric acid, naphthalene dicarboxylic acid, pyroglutamic acid, acetic acid, 1-naphthylcarboxylic acid, 2-naphthylcarboxylic acid, 1,8-naphthyl dicarboxylic acid, aconitic acid, carboxycinnamic acid, triazole dicarboxylic acid, gluconic acid, 4-carboxyphenyl boronic acid, butane disulfonic acid, and maleic acid.

21. A compound comprising:

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

(2) an oligopeptide of the formula (AA)_n-AA³-AA²-AA¹, wherein;

AA¹ is selected from the group consisting of Leucine, Phenylalanine, Isoleucine, Alanine, Glycine, Tyrosine, 2-Naphthylalanine, Seine, Proline, and β

-Alanine,

AA² represents any amino acid,

AA³ represents isoleucine,

n is 0 or 1, and when n is 1, then (AA)_n is AA⁴ which represents any amino acid,

(3) a stabilizing group, the stabilizing group selected from

(a) a dicarboxylic or higher order carboxylic acid,

(b) a non-genetically encoded amino acid having four or more carbons, or

(c) one of aspartic acid linked to the oligopeptide at the β

-carboxy group of the aspartic acid or glutamic acid linked to the oligopeptide at the γ

-carboxy group of the glutamic acid, and

(4) optionally, a linker group not cleavable by a trouase,

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 compound by enzymes present in whole blood, and

wherein the compound is cleavable by an enzyme associated with the target cell, the enzyme associated with the target cell being other than a trouase

or a salt of said compound.

22. The compound of claim 21 wherein the compound is resistant to cleavage by a trouase.

23. The compound of claim 21 wherein the stabilizing group is selected from the group consisting of:

succinic acid, adipic acid, glutaric acid, phthalic acid, diglycolic acid, fumaric acid, naphthalene dicarboxylic acid, pyroglutamic acid, 1,8-naphthyl dicarboxylic acid, aconitic acid, carboxycinnamic acid, triazole dicarboxylic acid, and maleic acid.

24. The compound of claim 21 wherein the stabilizing group is selected from the group consisting of:

Thiazolidine-4-carboxylic acid, 2-Thienylalanine, 2-Naphthylalanine, D-Leucine, D-Methionine, D-Phenylalanine, 3-Amino-3-phenyipropionic acid, γ

-Aminobutyric acid, 3-amino-4,4-diphenylbutyric acid, Tetrahydroisoquinoline-3-carboxylic acid, 4-Aminomethylbenzoic acid, and Aminoisobutyric acid.

25. A pharmaceutical composition comprising

(1) a compound including

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

(b) an oligopeptide of the formula (AA)_n-AA³-AA²-AA¹, wherein:

AA¹ is selected from the group consisting of Leucine, Phenylalanine, Isoleucine, Alanine, Glycine, Tyrosine, 2-Naphthylalanine, Serine, Proline, and β

-Alanine,

AA² represents any amino acid,

AA³ represents isoleucine,

n is0 or 1, and when n is 1, then (AA)_n is AA⁴ which represents any amino acid,

(c) a negatively charged stabilizing group, and

(d) optionally, a linker group not cleavable by TOP,

wherein the oligopeptide is directly linked to the stabilizing group at a first attachment site of the oligopeptide and AA¹ of 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 compound by enzymes present in whole blood, and

wherein the compound is cleavable by an enzyme associated with the target cell, the enzyme associated with the target cell being other than TOP, or a salt of said compound and

(2) a pharmaceutically acceptable carrier.

27. The pharmaceutical composition of claim 25 or 26, wherein the compound is resistant to cleavage by TOP.

26. A pharmaceutical composition comprising

(1) a compound including

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

(b) an oligopeptide of the formula (AA)_n-AA³-AA²-AA¹, wherein:

AA¹ is selected from the group consisting of Leucine, Phenylalanine, Isoleucine, Alanine, Glycine, Tyrosine, 2-Naphthylalanine, Serine, Proline, and β

-Alanine,

AA² is selected from the group consisting of Alanine, Leucine, Glycine, Serine, 3-Pyridylalanine, 2-Thienylalanine, Tyrosine, N-Methylalanine, Aminoisobutyric Acid, Threonine, and Phenylalanine,

AA³ represents isoleucine, and

n is0 or 1, and when n is 1, then (AA)_n is AA⁴ which represents any amino acid,

(c) a neutral stabilizing group or a negatively charged stabilizing group, and

(d) optionally, a linker group not cleavable by TOP,

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 compound by enzymes present in whole blood, and

wherein the compound is cleavable by an enzyme associated with the target cell, the enzyme associated with the target cell being other than TOP, or a salt of said compound, and

(2) a pharmaceutically acceptable carrier.

28. A method for treating a patient, the method comprising:

administering a therapeutically effective amount of a compound comprising;

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

(2) an oligopeptide of the formula (AA)_n-AA³-AA²-AA¹, wherein;

AA¹ is selected from the group consisting of Leucine, Phenylalanine, Isoleucine, Alanine, Glycine, Tyrosine, 2-Naphthylalanine, Serine, Proline, and β

-Alanine,

AA² represents any amino acid,

AA³ represents isoleucine,

n is 0 or 1, and when n is 1 then (AA)_n is AA⁴ which represents any amino acid,

(3) a negatively charged stabilizing group, and

(4) optionally, a linker group not cleavable by TOP, or a salt of said compound

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 compound by enzymes present in whole blood, and

wherein the compound is cleavable by an enzyme associated with the target cell, the enzyme associated with the target cell being other than TOP, such that the patient is treated.

30. The method of claim 28 or 29, wherein the compound is resistant to cleavage by TOP.

31. The method of claim 28 or 29, wherein the compound is administered intravenously.

32. The method of claim 28 or 29, wherein the patient is treated for a medical condition selected from cancer, neoplastic diseases, tumors, inflammatory diseases, or infectious diseases.

29. A method for treating a patient, the method comprising:

administering a therapeutically effective amount of a compound comprising;

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

(2) an oligopeptide of the formula (AA)_n-AA³-AA²-AA¹, wherein;

AA¹ is selected from the group consisting of Leucine, Phenylalanine, Isoleucine, Alanine, Glycine, Tyrosine, 2-Naphthylalanine, Serine, Proline, and β

-Alanine,

AA² is selected from the group consisting of Alanine, Leucine, Glycine, Serine, 3-Pyridylalanine, 2-Thienylalanine, Tyrosine, N-Methylalanine, Aminoisobutyric Acid, Threonine, and Phenylalanine,

AA³ represents isoleucine, and

n is 0 or 1, and when n is 1, then (AA)_n is AA⁴ which represents any amino acid,

(3) a neutral stabilizing group or a negatively charged stabilizing group, and

(4) optionally, a linker group not cleavable by TOP,

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 compound by enzymes present in whole blood, and

wherein the compound is cleavable by an enzyme associated with the target cell, the enzyme associated with the target cell being other than TOP, or a salt thereof, such that the patient is treated.

33. An article of manufacture for diagnosis or assay comprising:

(1) a compound comprising;

(a) a marker,

(b) an oligopeptide of the formula (AA)_n-AA³-AA²-AA¹, wherein;

AA¹ is selected from the group consisting of Leucine, Phenylalanine, Isoleucine, Alanine, Glycine, Tyrosine, 2-Naphthylalanine, Serine, Proline, and β

-Alanine,

AA² is selected from the group consisting of;

Alanine, Leucine, Glycine, Serine, Tyrosine, 3-Pyridylalanine, 2-Thienylalanine, N-Methylalanine, Aminoisobutyric Acid, Threonine, and Phenylalanine,

AA³ represents isoleucine,

n is 0 or 1, and when n is 1 then (AA)_n is AA⁴ which represents any amino acid,

(c) a stabilizing group, and

(d) optionally, a linker group not cleavable by TOP, wherein the oligopeptide is directly linked to the stabilizing group at a first attachment site of the oligopeptide and AA¹ of the oligopeptide is directly linked to the marker or indirectly linked through the linker group to the marker at a second attachment site of the oligopeptide, wherein the stabilizing group hinders cleavage of the compound by enzymes present in whole blood, and wherein the compound is cleavable by an enzyme associated with a target cell, the enzyme associated with the target cell being other than TOP, and further wherein the compound is resistant to cleavage by TOP, and

(2) at least one reagent useful in the detection of said marker.

34. The article of manufacture according to claim 33 wherein AA⁴ is selected from β

-Alanine, Thiazolidine-4-carboxylic acid, 2-Thienylalanine, 2-Naphthylalanine, D-Alanine, D-Leucine, D-Methionine, D-Phenylalanine, 3-Amino-3-phenylpropionic acid, γ

-Aminobutyric acid, 3-amino-4,4-diphenylbutyric acid, Tetrahydroisoquinoline-3-carboxylic acid, 4-Aminomethylbenzoic acid, and Aminoisobutyric acid.

35. The article of manufacture according to claim 33 wherein the oligopeptide is β

Ala-Ile-Ala-Leu (SEQ ID NO;

     8).

36. The article of manufacture according to claim 33 wherein the stabilizing group is negatively charged or neutral.

37. The article of manufacture according to claim 33 wherein the stabilizing group is a dicarboxylic or higher order carboxylic acid.

38. The article of manufacture according to claim 33 wherein the stabilizing group is selected from the group consisting of succinic acid, adipic acid, glutaric acid, phthalic acid, diglycolic acid, fumaric acid, naphthalene dicarboxylic acid, pyroglutamic acid, acetic acid, 1-naphthylcarboxylic acid, 2-naphthylcarboxylic acid, 1,8-naphthyl dicarboxylic acid, aconitic acid, carboxycinnamic acid, triazole dicarboxylic acid, gluconic acid, 4-carboxyphenyl boronic acid, polyethylene glycolic acid, butane disulfonic acid, maleic acid, nipecotic acid, and isonipecotic acid.

39. The article of manufacture according to claim 33 wherein the stabilizing group is a non-genetically encoded amino acid.

40. The article of manufacture according to claim 33 wherein the stabilizing group is selected from the group consisting of:

β

-Alanine, Thiazolidine-4-carboxylic acid, 2-Thienylalanine, 2-Naphthylalanine, D-Alanine, D-Leucine, D-Methionine, D-Phenylalanine, 3-Amino-3-phenylpropionic acid, γ

-Aminobutyric acid, 3-amino-4,4-diphenylbutyric acid, Tetrahydroisoquinoline-3-carboxylic acid, 4-Aminomethylbenzoic acid, and Aminoisobutyric acid.

41. The article of manufacture according to claim 33 wherein the stabilizing group is one of aspartic acid linked to the oligopeptide at the β

-carboxy group of the aspartic acid or glutamic acid linked to the oligopeptide at the γ

-carboxy group of the glutamic acid.