Compositions and Methods for Delivery of Antitumor Agents

• US 20080267981A1
• Filed: 06/29/2005
• Published: 10/30/2008
• Est. Priority Date: 06/30/2004
• Status: Abandoned Application

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Boger, Dale L., Janda, Kim D., Wirsching, Peter
• Application Number:
  US11/630,101
• Publication Number:
  US 20080267981A1
• Time in Patent Office: Days
• US Class Current: 424/181.1
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07D 403/06 linked by a carbon chain containing only aliphatic carbon atoms
• CPC Class Codes: C07D 403/14 containing three or more hetero rings
• CPC Class Codes: C07D 487/04 Ortho-condensed systems
• CPC Class Codes: C07D 487/10 Spiro-condensed systems

Abstract

Methods for treating a neoplastic disease with an antibody-cytotoxin conjugate molecule, methods of synthesizing an antibody-cytotoxin conjugate molecule are provided. Compounds that are useful as antibody-cytotoxin conjugate molecule or useful in the synthesis of these molecules are also provided.

Claims

1. A method for treating a neoplastic disease in a mammal comprising:

providing an antibody-cytotoxin conjugate with an acid-stable covalent linkage between an antibody and a cytotoxin, and

administering said antibody-cytotoxin conjugate to the mammal so that said conjugate is internalized within a cell of the mammal to treat said neoplastic disease.

2. The method of claim 1 wherein said cytotoxin is an antitumor antibiotic, duocarmycin, duocarmycin A, duocarmycin SA, or an analog thereof.

3. The method of claim 1 wherein said acid-stable linkage is an amide linkage.

4. The method of claim 3 wherein said amide linkage is an N-substituted amide linkage.

5. The method of claim 1 wherein the antibody portion of said conjugate specifically binds to an activated integrin receptor.

6. The method of claim 5 wherein said activated integrin receptor is differentially produced on a cell in a metastatic state as compared to a similar, non-metastatic cell.

7. The method of claim 6 wherein said activated integrin receptor is an α

₃β

₁ integrin receptor or an α

vβ

3 integrin receptor.

8. The method of claim 1 wherein the antibody portion of said conjugate is a single chain Fv antibody.

9. The method of claim 1 wherein said neoplastic disease is selected from solid tumor, hematological malignancy, leukemia, colorectal cancer, benign or malignant breast cancer, uterine cancer, uterine leiomyomas, ovarian cancer, endometrial cancer, polycystic ovary syndrome, endometrial polyps, squamous cell carcinoma, squamous cell carcinoma of the head and neck, hepatocellular carcinoma, intrahepatic metastasis of hepatocellular carcinoma, prostate cancer, prostatic hypertrophy, pituitary cancer, adenomyosis, adenocarcinoma, pancreatic adenocarcinoma, meningioma, melanoma, bone cancer, multiple myeloma, CNS cancer, glioma, or astroblastoma.

10. A method for treating a neoplastic disease in a mammal comprising:

providing an antibody-cytotoxin conjugate with an acid-labile covalent linkage between an antibody and a cytotoxin, and

administering said antibody-cytotoxin conjugate to the mammal so that said conjugate is internalized within a cell of the mammal to treat said neoplastic disease.

11. The method of claim 10 wherein said cytotoxin is an antitumor antibiotic, duocarmycin, duocarmycin A, duocarmycin SA, or an analog thereof.

12. The method of claim 10 wherein said acid-labile covalent linkage is a hydrazone linkage.

13. The method of claim 12, further comprising administering said antibody-antitumor antibiotic conjugate so that said conjugate is internalized within a cell of the mammal by cleavage of an acid-labile hydrazone linkage.

14. The method of claim 10 wherein said antibody specifically binds to an activated integrin receptor.

15. The method of claim 14 wherein said activated integrin receptor is differentially produced on a cell in a metastatic state as compared to a similar, non-metastatic cell.

16. The method of claim 15 wherein said activated integrin receptor is an α

₃β

₁ integrin receptor or an α

vβ

3 integrin receptor.

17. The method of claim 10 wherein the antibody portion of said conjugate is a single chain Fv antibody.

18. The method of claim 10 wherein said neoplastic disease is selected from solid tumor, hematological malignancy, leukemia, colorectal cancer, benign or malignant breast cancer, uterine cancer, uterine leiomyomas, ovarian cancer, endometrial cancer, polycystic ovary syndrome, endometrial polyps, squamous cell carcinoma, squamous cell carcinoma of the head and neck, hepatocellular carcinoma, intrahepatic metastasis of hepatocellular carcinoma, prostate cancer, prostatic hypertrophy, pituitary cancer, adenomyosis, adenocarcinoma, pancreatic adenocarcinoma, meningioma, melanoma, bone cancer, multiple myeloma, CNS cancer, glioma, or astroblastoma.

19. A method of synthesizing an antibody-cytotoxin conjugate molecule comprising:

introducing into a single vessel an antibody, a thiolating reagent, and a maleimide-derivatized cytotoxin;

contacting said antibody with said thiolating reagent to form a thiolated antibody; and

contacting said thiolated antibody with said maleimide-derivatized cytotoxin to form an antibody-cytotoxin conjugate molecule.

20. The method of claim 19 wherein said maleimide-derivatized cytotoxin comprises an acid-labile hydrazone linkage between maleimide and the cytotoxin.

21. The method of claim 19 wherein said maleimide-derivatized cytotoxin comprises an amide bond linkage between maleimide and the cytotoxin.

22. The method of claim 19, wherein said cytotoxin is an antitumor antibiotic, duocarmycin, duocarmycin A, duocarmycin SA, or an analog thereof.

23. The method of claim 22 wherein said antitumor antibiotic is a carbonyl-substituted CBI-indole analog of duocarmycin SA.

24. The method of claim 22 wherein said antitumor antibiotic is an amide-substituted CBI-indole analog of duocarmycin SA.

25. The method of claim 23 wherein said maleimide-derivatized cytotoxin is 1-[3-(N′

-{1-[2-(1-Chloromethyl-5-hydroxy-1,2-dihydro-3H-benzo[e]indole-3-carbonyl)-1H-indol-5-yl]-ethylidene}-hydrazino)-3-oxo-1-propyl]maleimide.

26. The method of claim 24 wherein said maleimide-derivatized cytotoxin is 3-[5-[1-{3-[3-(2,5-dioxo-2,5-dihydropyrrol-1-yl)propionylamino]propyl}indole-2-carbonyl]aminoindole-2-carbonyl]-(1-chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indole.

27. The method of claim 19, wherein said thiolating reagent is 2-iminothiolane.

28. The method of claim 19, wherein said antibody is a single chain Fv antibody.

29. A compound 3-[5-(1-(3-aminopropyl)indole-2-carbonyl)aminoindole-2-carbonyl]-1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benz[e]indole.

30. A compound of Formula I:

31. The compound of claim 30 wherein R₂ is C₁ to C₆ alkyl.

32. The compound of claim 30, wherein halogen is Cl, Br, or F.

35. A pharmaceutical composition comprising at least one pharmaceutically acceptable carrier or excipient and an effective amount of the compound of claim 30, wherein the maleimide moiety is conjugated to a single chain Fv antibody.

36. The composition of claim 35 wherein said single chain Fv antibody is capable of binding to an integrin receptor.

37. The composition of claim 36 wherein said integrin receptor is an-α

₃β

₁ integrin receptor or an α

vβ

3 integrin receptor.

38. A method comprising administering to a mammal the composition of claim 36.

39. A method for alleviating a disease state in a mammal believed to be responsive to treatment with an antibody conjugated to a amide-substituted CBI-indole analog of duocarmycin SA comprising the step of administering to the mammal a therapeutic amount of the composition of claim 36.

40. The method of claim 36 wherein said compound is 3-[5-(1-(3-aminopropyl)indole-2-carbonyl)aminoindole-2-carbonyl]-1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benz[e]indole.

41. The method of claim 39, wherein said disease state is neoplastic disease.

42. The method of claim 40 wherein the neoplastic disease is selected from solid tumor, hematological malignancy, leukemia, colorectal cancer, benign or malignant breast cancer, uterine cancer, uterine leiomyomas, ovarian cancer, endometrial cancer, polycystic ovary syndrome, endometrial polyps, squamous cell carcinoma, squamous cell carcinoma of the head and neck, hepatocellular carcinoma, intrahepatic metastasis of hepatocellular carcinoma, prostate cancer, prostatic hypertrophy, pituitary cancer, adenomyosis, adenocarcinoma, pancreatic adenocarcinoma, meningioma, melanoma, bone cancer, multiple myeloma, CNS cancer, glioma, or astroblastoma.

33. The compound 3-[5-(1-(3-aminopropyl)indole-2-carbonyl)aminoindole-2-carbonyl]-1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benz[e]indole.

34. The compound 3-[5-[1-{3-[3-(2,5-dioxo-2,5-dihydropyrrol-1-yl)propionylamino]propyl}indole-2-carbonyl]aminoindole-2-carbonyl]-(1-chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indole.

43. A compound of Formula II:

45. A pharmaceutical composition comprising at least one pharmaceutically acceptable carrier or excipient and an effective amount of the compound of claim 43, wherein the maleimide moiety is conjugated to a single chain Fv antibody.

46. The composition of claim 45 wherein said single chain Fv antibody is an antibody to an integrin receptor.

47. The composition of claim 46 wherein said integrin receptor is an-α

₃β

₁ integrin receptor or an α

vβ

3 integrin receptor.

48. A method comprising administering to a mammal the composition of claim 46.

49. A method for alleviating a disease state in a mammal believed to be responsive to treatment with an antibody conjugated to a carbonyl-substituted CBI-indole analog of duocarmycin SA comprising the step of administering to the mammal a therapeutic amount of the composition of claim 46.

50. The method of claim 49 wherein said disease state is neoplastic disease.

51. The method of claim 50 wherein the neoplastic disease is selected from solid tumor, hematological malignancy, leukemia, colorectal cancer, benign or malignant breast cancer, uterine cancer, uterine leiomyomas, ovarian cancer, endometrial cancer, polycystic ovary syndrome, endometrial polyps, squamous cell carcinoma, squamous cell carcinoma of the head and neck, hepatocellular carcinoma, intrahepatic metastasis of hepatocellular carcinoma, prostate cancer, prostatic hypertrophy, pituitary cancer, adenomyosis, adenocarcinoma, pancreatic adenocarcinoma, meningioma, melanoma, bone cancer, multiple myeloma, CNS cancer, glioma, or astroblastoma.

44. A compound 1-[3-(N′

-{1-[2-(1-Chloromethyl-5-hydroxy-1,2-dihydro-3H-benzo[e]indole-3-carbonyl)-1H-indol-5-yl]-ethylidene}-hydrazino)-3-oxo-1-propyl]maleimide.