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:
each AA independently represents an amino acid, n is 0 or 1, and when n is 1, then (AA)n is AA4 which represents any amino acid, AA3 represents isoleucine, AA2 represents any amino acid, and AA1 represents any amino acid, (3) a 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.
- 2. The compound of claim 1 wherein the compound is resistant to cleavage by TOP.
- 3. The compound of claim 1 wherein AA1 of the oligopeptide is selected from Leucine, Phenylalanine, Isoleucine, Alanine, Glycine, Tyrosine, 2-Naphthylalanine, Serine, Proline, and β-Alanine.
- 4. The compound of claim 1 wherein AA2 of the oligopeptide is selected from Alanine, Leucine, Glycine, Serine, Tyrosine, 3-Pyridylalanine, 2-Thienylalanine, Tyrosine, N-Methylalanine, Aminoisobutyric Acid, Threonine, and Phenylalanine.
- 5. The compound of claim 1 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-Aminomethylbenzoic acid, and Aminoisobutyric acid.
- 6. The compound of claim 1 wherein the oligopeptide is selected from βAla-Ile-Ala-Phe (SEQ ID NO: 1); βAla-Ile-Ala-Ile (SEQ ID NO: 2); Tic-Ile-Ala-Leu (SEQ ID NO: 3); Thi-Ile-Ala-Leu (SEQ ID NO: 4); Nal-Ile-Ala-Leu (SEQ ID NO: 5); Amb-Ile-Ala-Leu (SEQ ID NO: 6); Aib-Ile-Ala-Leu (SEQ ID NO: 7); βAla-Ile-Ala-Leu (SEQ ID NO: 8); Thi-Ile-Aib-Leu (SEQ ID NO: 9); Nal-Ile-Aib-Leu (SEQ ID NO: 10); βAla-Ile-Aib-Leu (SEQ ID NO: 11); Amb-Ile-Aib-Leu (SEQ ID NO: 12); Aib-Ile-Aib-Leu (SEQ ID NO: 13); βAla-Ile-Gly-Phe (SEQ ID NO: 14); βAla-Ile-Gly-Ile (SEQ ID NO: 15); Tic-Ile-Gly-Leu (SEQ ID NO: 16); Thi-Ile-Gly-Leu (SEQ ID NO: 17); Nal-Ile-Gly-Leu (SEQ ID NO: 18); βAla-Ile-Gly-Leu (SEQ ID NO: 19); Amb-Ile-Gly-Leu (SEQ ID NO: 20); Aib-Ile-Gly-Leu (SEQ ID NO: 21); βAla-Ile-Thr-Ile (SEQ ID NO: 22); βAla-Ile-Tyr-Ile (SEQ ID NO: 23); βAla-Ile-Ala-Gly (SEQ ID NO: 24); Ile-Ala-Leu (SEQ ID NO: 25); Ile-N(Me)Ala-Leu (SEQ ID NO: 26); Ile-Ala-Phe (SEQ ID NO: 27); Ile-Ala-Ile (SEQ ID NO: 28); Ile-Aib-Leu (SEQ ID NO: 29); Ile-Gly-Phe (SEQ ID NO: 30); Ile-Gly-Ile (SEQ ID NO: 31); Ile-Gly-Leu (SEQ ID NO: 32); Ile-Thr-Ile (SEQ ID NO: 33); and Ile-Ala-Gly (SEQ ID NO: 34); βAla-Ile-Tyr-Leu (SEQ ID NO: 35); βAla-Ile-Tyr-Gly (SEQ ID NO: 36).
- 7. The compound of claim 1 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, 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.
- 9. The compound of claim 1 wherein the stabilizing group is a non-genetically encoded amino acid.
- 10. The compound of claim 9 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.
- 11. The compound of claim 1 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.
- 12. The compound of claim 1 wherein the stabilizing group is negatively charged or neutral.
- 13. The compound of claim 1 wherein the enzyme associated with the target cell cleaves the linkage between AA1 and AA2 of the oligopeptide.
- 14. The compound of claim 1 wherein the oligopeptide is directly linked to the therapeutic agent.
- 15. The compound of claim 1 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 sulfhydryl group.
- 16. The compound of claim 1 wherein the therapeutic agent is selected from the group consisting of Alkylating Agents, Antiproliferative agents, Tubulin Binding agents, Vinca Alkaloids, Enediynes, Podophyllotoxins or Podophyllotoxin derivatives, the Pteridine family of drugs, Taxanes, Anthracyclines, Dolastatins, Topoiosomerase inhibitors, Platinum coordination-complex chemotherapeutic agents, and Maytansinoids.
- 17. The compound of claim 1 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, Mitomycin C, Bleomycin, Melphalan, Chloroquine, Cyclosporin A, Maytansine, a derivative of any of the foregoing and an analog any of the foregoing.
- 18. The compound of claim 1 wherein the therapeutic agent has an intracellular active site.
- 19. The compound of claim 1 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 enzyme associated with the target cell than prior to cleavage by the enzyme associated with the target cell, the active portion including at least the therapeutic agent.
- 20. The compound of claim 19 wherein the active portion of the prodrug consists of the therapeutic agent.
- 21. The compound of claim 19 wherein the active portion of the prodrug includes the therapeutic agent and at least the linker group.
- 22. The compound of claim 19 wherein the active portion of the prodrug includes the therapeutic agent and AA1 of the oligopeptide.
- 23. The compound of claim 1 wherein the compound is selected from the group consisting of: Suc-βAla-Ile-Ala-Phe-Dnr (SEQ ID NO: 49), Suc-βAla-Ile-Ala-Ile-Dnr (SEQ ID NO: 50), Suc-Tic-Ile-Ala-Leu-Dnr (SEQ ID NO: 51), Suc-Thi-Ile-Ala-Leu-Dnr (SEQ ID NO: 52), Suc-Nal-Ile-Ala-Leu-Dnr (SEQ ID NO: 53), Suc-βAla-Ile-Ala-Leu-Dnr (SEQ ID NO: 43), Suc-Amb-Ile-Ala-Leu-Dnr (SEQ ID NO: 54), Suc-Aib-Ile-Ala-Leu-Dnr (SEQ ID NO: 55), Suc-βAla-Ile-Ala-Leu-Dox (SEQ ID NO: 39), Suc-Thi-Ile-Aib-Leu-Dnr (SEQ ID NO: 56), Suc-Nal-Ile-Aib-Leu-Dnr (SEQ ID NO: 57), Suc-βAla-Ile-Aib-Leu-Dnr (SEQ ID NO: 58), Suc-Amb-Ile-Aib-Leu-Dox (SEQ ID NO: 59), Suc-Aib-Ile-Aib-Leu-Dnr (SEQ ID NO: 60), Suc-βAla-Ile-Gly-Phe-Dnr (SEQ ID NO: 61), Suc-βAla-Ile-Gly-Ile-Dnr (SEQ ID NO: 62), Suc-Tic-Ile-Gly-Leu-Dnr (SEQ ID NO: 63), Suc-Thi-Ile-Gly-Leu-Dnr (SEQ ID NO: 64), Suc-Nal-Ile-Gly-Leu-Dnr (SEQ ID NO: 65), Suc-βAla-Ile-Gly-Leu-Dnr (SEQ ID NO: 66), Suc-Amb-Ile-Gly-Leu-Dnr (SEQ ID NO: 67), Suc-Aib-Ile-Gly-Leu-Dnr (SEQ ID NO: 68), Suc-βAla-Ile-Thr-Ile-Dnr (SEQ ID NO: 69), Suc-βAla-Ile-Tyr-Ile-Dnr (SEQ ID NO: 70), Suc-βAla-Ile-Tyr-Leu-Dnr (SEQ ID NO: 71), Suc-βAla-Ile-Tyr-Gly-Dox (SEQ ID NO: 72), Suc-βAla-Ile-Ala-Gly-Dox (SEQ ID NO: 73), Suc-Ile-Ala-Leu-Dox, Suc-Ile-N(Me)Ala-Leu-Dox, and Suc-Ile-Ala-Gly-Dox.
- 24. The compound of claim 1 wherein the target cell is a tumor cell or inflammatory cell.
- 25. The compound of claim 1 wherein the enzyme associated with the target cell is present in the extracellular vicinity of the target cell for the therapeutic agent.
- 26. 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:
each AA independently represents an amino acid, n is 0 or 1, and when n is 1, then (AA)n is AA4 which represents any amino acid, AA3 represents isoleucine, AA2 represents any amino acid, and AA1 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, 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.
- 27. The compound of claim 26 wherein the compound is resistant to cleavage by a trouase.
- 28. The compound of claim 26 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, and maleic acid.
- 29. 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:
each AA independently represents an amino acid, n is 0 or 1, and when n is 1, then (AA)n is AA4 which represents any amino acid, AA3 represents isoleucine, AA2 represents any amino acid, and AA1 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 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, and (2) a pharmaceutically acceptable carrier.
- 30. The compound of claim 29 wherein the compound is resistant to cleavage by TOP.
- 31. 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:
each AA independently represents an amino acid, n is 0 or 1, and when n is 1, then (AA)n is AA4 which represents any amino acid, AA3 represents isoleucine, AA2 represents any amino acid, and AA1 represents any amino acid, (3) a 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.
- 32. The method of claim 31 wherein the compound is resistant to cleavage by TOP.
- 33. The method of claim 31 wherein the compound is administered intravenously.
- 34. The method of claim 31 wherein the patient is treated for a medical condition selected from cancer, neoplastic diseases, tumors, inflammatory diseases, or infectious diseases.
- 35. A method of designing a prodrug for administration to a patient, the method comprising:
(1) identifying an oligopeptide of the formula (AA)n-AA3-AA2-AA1, wherein:
each AA independently represents an amino acid, n is 0 or 1, and when n is 1, then (AA)n is AA4 which represents any amino acid, AA3 represents isoleucine, AA2 represents any amino acid, and AA1 represents any amino acid, (2) linking the oligopeptide at a first attachment site of the oligopeptide to a stabilizing group, and (3) directly or indirectly linking the oligopeptide to a therapeutic agent at a second attachment site of the oligopeptide, wherein steps (2) and (3) may be performed in any order or concurrently and further wherein a conjugate is formed by performance of steps (1) through (3), (4) testing if the conjugate is cleavable by TOP, (5) testing if the conjugate is stable in whole blood, and (6) selecting the conjugate as a prodrug if the conjugate is resistant to cleavage by TOP and stable in whole blood.
- 36. The method of claim 35 wherein the first attachment site is the N-terminus of the oligopeptide.
- 37. The method of claim 35 wherein the second attachment site is the C-terminus of the oligopeptide.
- 38. The method of claim 35 wherein the first attachment site is the C-terminus of the oligopeptide.
- 39. The method of claim 35 wherein the second attachment site is the N-terminus of the oligopeptide.
- 40. A prodrug designed by the method of claim 35.
- 41. A method for decreasing toxicity of a therapeutic agent wherein the therapeutic agent is intended for administration to a patient, the method comprising:
forming a prodrug form of the therapeutic agent by covalently linking an oligopeptide 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, said oligopeptide having a formula (AA)n-AA3-AA2-AA1, wherein:
each AA independently represents an amino acid, n is 0 or 1, and when n is 1, then (AA)n is AA4 which represents any amino acid, AA3 represents isoleucine, AA2 represents any amino acid, and AA1 represents any amino acid, and further wherein the oligopeptide is resistant to cleavage by a trouase, the prodrug being cleavable by an enzyme associated with a target cell, the enzyme associated with the target cell being other than a trouase, and further wherein the prodrug is resistant to cleavage by a trouase, whereby the prodrug form of the therapeutic agent provides for decreased toxicity of the therapeutic agent when administered to the patient.
- 42. The method of claim 41 wherein the prodrug allows for administration of an increased dosage of the therapeutic agent in prodrug form to the patient relative to the dosage of the therapeutic agent in unconjugated form.
- 43. 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:
each AA independently represents an amino acid, n is 0 or 1, and when n is 1, then (AA)n is AA4 which represents any amino acid, AA3 represents isoleucine, AA2 represents any amino acid, and AA1 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 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, and further wherein the compound is resistant to cleavage by TOP, and (2) optionally at least one reagent useful in the detection of said marker.
- 44. In a method of making a prodrug, a method of removing free therapeutic agent comprising:
(1) coupling an optionally protected stabilizing group-oligopeptide conjugate with the free therapeutic agent, (2) contacting the reactants of step (1) with a polymeric resin to bind free therapeutic agent remaining after step (1) and to form a therapeutic agent-polymeric resin complex, and (3) removing the therapeutic agent-polymeric resin complex.
- 45. The method of claim 44 wherein the polymeric resin is polystyrene methylisocyanate or polystyrene sulfonyl chloride.
- 46. The method of claim 44 wherein the optionally protected stabilizing group-oligopeptide conjugate includes an oligopeptide of the formula (AA)n-AA3-AA2-AA1, wherein:
each AA independently represents an amino acid, n is 0 or 1, and when n is 1, then (AA)n is AA4 which represents any amino acid, AA3 represents isoleucine, AA2 represents any amino acid, and AA1 represents any amino acid.
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application is related to U.S. Ser. No. 60/211,686, filed Jun. 14, 2000.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
PCT/US01/18857 |
6/11/2001 |
WO |
|