COMPOSITIONS AND METHODS FOR ENHANCING DELIVERY OF THERAPEUTIC AGENTS TO CELLS

• US 20010006946A1
• Filed: 07/08/1998
• Published: 07/05/2001
• Est. Priority Date: 01/08/1996
• Status: Active Grant

• Current Assignee: Canji Inc. (Merck & Co., Inc.), Schering Corporation (Merck & Co., Inc.)
• Original Assignee: Canji Inc. (Merck & Co., Inc.)
• Inventors: ENGLER, HEIDRUN, NAGABHUSHAN, TATTANAHALLI L.
• Granted Patent: US 6,392,069 B2
• Time in Patent Office: Days
• US Class Current: 514/44
• CPC Class Codes: A61K 31/56 Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
• CPC Class Codes: A61K 31/7008 Compounds having an amino group directly attached to a carbon atom of the saccharide radical, e.g. D-galactosamine, ranimustine
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07J 41/0061 one of the carbon atoms being part of an amide group
• CPC Class Codes: C07J 9/005 containing a carboxylic function directly attached or attached by a chain containing only carbon atoms to the cyclopenta[a]hydrophenanthrene skeleton
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

This invention provides methods and compositions for enhancing transfer of an agent into a cell. The agents can include polypeptides, polynucleotides such as genes and antisense nucleic acids, and other molecules. In some embodiments, the agents are modulating agents that can modulate a cellular activity or function when introduced into the cell. The methods and compositions are useful for introducing agents into individual cells, as well as cells that are present as a tissue or organ.

Claims

1. A method of delivering an agent to cells, the method comprising administering the agent to the cells in a composition comprising a delivery enhancing compound of Formula I:

wherein;

m and n are the same or different and each is an integer from 2-8;

R is a cationic group or

X₁ is a cholic acid group or deoxycholic acid group; and

X₂ and X₃ are each independently selected from the group consisting of a cholic acid group, a deoxycholic acid group, and a saccharide group;

wherein at least one of X₂ and X₃ is a saccharide group when R is

2. The method of

claim 1

, wherein the amount of the agent delivered to the cells in the presence of the delivery enhancing agent is increased relative to the amount of the agent delivered to the cells when the agent is administered in the absence of the delivery enhancing compound.

3. The method of

claim 1

, wherein the agent is a therapeutic agent.

4. The method of

claim 1

, wherein the concentration of the delivery enhancing compound is about 0.002 to about 2 mg/ml.

5. The method of

claim 4

, wherein the concentration of the delivery enhancing compound is about 0.02 to about 2 mg/ml.

6. The method of

claim 5

, wherein the concentration of the delivery enhancing compound is about 0.2 to 2 mg/ml.

7. The method of

claim 1

, wherein the cells are provided as a tissue.

8. The method of

claim 1

, wherein the tissue is an organ.

9. The method of

claim 1

, wherein the administration is by intravesical administration.

10. The method of

claim 1

, wherein the agent is a protein.

11. The method of

claim 1

, wherein the agent is a gene.

12. The method of

claim 11

, wherein the gene is administered in a vector.

13. The method of

claim 12

, wherein the vector is a viral vector.

14. The method of

claim 13

, wherein the viral vector is selected from the group consisting of an adenoviral vector, a retroviral vector, and an adeno-associated viral vector.

15. The method of

claim 13

, wherein the viral vector is administered as a suspension containing from about 1×

10⁸ particles/ml to about 5×

10¹¹ particles/ml of the viral vector.

16. The method of

claim 15

, wherein suspension contains from about 1×

10⁹ particles/ml to about 1×

10¹¹ particles/ml of the viral vector.

17. The method of

claim 11

, wherein the gene is a therapeutic gene.

18. The method of

claim 17

, wherein the therapeutic gene is a tumor suppressor gene.

19. The method of

claim 18

, wherein the tumor suppressor gene is p53.

20. The method of

claim 18

, wherein the tumor suppressor gene is a retinoblastoma gene.

21. The method of

claim 20

, wherein the retinoblastoma tumor suppressor gene encodes full length RB protein.

22. The method of

claim 20

, wherein the retinoblastoma tumor suppressor gene encodes p56^RB.

23. The method of

claim 17

, wherein the cells are cancer cells.

24. The method of

claim 23

, wherein the cancer cells are bladder cancer cells.

25. The method of

claim 23

, wherein the cancer cells are provided as a tissue.

26. The method of

claim 1

, wherein the delivery-enhancing compound is administered prior to administration of the agent.

27. The method of

claim 1

, wherein the delivery enhancing compound is administered with the agent.

28. A composition for delivering an agent to cells, the composition comprising the agent and a delivery enhancing compound of Formula I:

wherein;

m and n are the same or different and each is an integer from 2-8;

R is a cationic group or

X₁ is a cholic acid group or deoxycholic acid group; and

X₂ and X₃ are each independently selected from the group consisting of a cholic acid group, a deoxycholic acid group, and a saccharide group;

wherein at least one of X₂ and X₃ is a saccharide group when R is

29. The composition according to

claim 28

, wherein the saccharide group comprises one or more pentose or hexose residues.

30. The composition according to

claim 29

, wherein the saccharide group is selected from the group consisting of pentose monosaccharide groups, hexose monosaccharide groups, pentose-pentose disaccharide groups, hexose-hexose disaccharide groups, pentose-hexose disaccharide groups, and hexose-pentose disaccharide groups.

31. The composition according to

claim 28

, wherein the saccharide group is a trisaccharide.

32. The composition according to

claim 28

, wherein the concentration of the delivery enhancing compound is about 0.002 to about 2 mg/ml.

33. The composition according to

claim 32

, wherein the concentration of the delivery enhancing compound is about 0.2 to 2 mg/ml.

34. The composition according to

claim 28

, wherein the agent modulates a biological process in a cell when the agent is present in the cell.

35. The composition according to

claim 34

, wherein the biological process is selected from the group consisting of cell growth, differentiation, proliferation, a metabolic or biosynthetic pathway, gene expression, a disease-associated process, and an immune response.

36. The composition according to

claim 28

, wherein the agent comprises a polynucleotide.

37. The composition according to

claim 36

, wherein the polynucleotide is selected from the group consisting of an antisense nucleic acid, a triplex-forming nucleic acid, and a nucleic acid that comprises a gene which encodes a polypeptide.

38. The composition according to

claim 37

, wherein the gene is a tumor suppressor gene.

39. The composition according to

claim 37

, wherein the tumor suppressor gene is selected from the group consisting of a retinoblastoma gene and a p53 gene.

40. The composition according to

claim 28

, wherein the composition further comprises a polymeric matrix.

41. The composition according to

claim 28

, wherein the composition further comprises a mucoadhesive.

42. A delivery enhancing compound having a Formula I:

wherein;

m and n are the same or different and each is an integer from 2-8;

R is a cationic group or

X₁ is a cholic acid group or deoxycholic acid group; and

X₂ and X₃ are each independently selected from the group consisting of a cholic acid group, a deoxycholic acid group, and a saccharide group;

wherein at least one of X₂ and X₃ is a saccharide group when R is

43. The compound of

claim 42

, wherein R is a cationic group selected from the group consisting of NMe₃⁻

and NH₃⁺.

44. The compound of

claim 42

, wherein the saccharide group comprises one or more pentose or hexose residues.

45. The compound of

claim 44

, wherein the saccharide group is selected from the group consisting of pentose monosaccharide groups, hexose monosaccharide groups, pentose-pentose disaccharide groups, hexose-hexose disaccharide groups, pentose-hexose disaccharide groups, and hexose-pentose disaccharide groups.

46. The compound of

claim 42

, wherein the saccharide group comprises between three and about eight monosaccharide residues.

47. The compound of

claim 46

, wherein the saccharide group is a trisaccharide.

48. The compound of

claim 42

, wherein at least one of X₂ and X₃ is a saccharide group.

49. The compound of

claim 42

, wherein m and n are each independently 2 or 3.

50. The compound of

claim 42

, wherein both X₁ and X₂ are both cholic acid groups and X₃ is a saccharide group.

51. The compound of

claim 42

, wherein the saccharide group is a hexose-hexose disaccharide group.

52. The compound of

claim 42

, wherein m and n are each 3, X₁ and X₂ are both cholic acid groups, and X₃ is a hexose monosaccharide group.

53. The compound of

claim 42

, wherein m and n are each 3, X₁ and X₃ are both cholic acid groups, and X₂ is a hexose monosaccharide group.

54. The compound of

claim 42

, wherein m and n are each 3, X₁ and X₂ are both cholic acid groups, and X₃ is a hexose-hexose disaccharide group.

55. The compound of

claim 42

, wherein m and n are each 3, X₁ and X₃ are both cholic acid groups, and X₂ is a hexose-hexose disaccharide group.

56. The compound according to

claim 42

, wherein the compound has a Formula III:

57. The compound according to

claim 42

, wherein the compound has a Formula IV:

58. The compound according to

claim 42

, wherein the compound has a Formula V:

59. A delivery enhancing compound of Formula II:

wherein X₁ and X₂ are selected from the group consisting of a cholic acid group and a deoxycholic acid group and X₃ is a saccharide group.

60. The compound according to

claim 59

, wherein both X₁ and X₂ are cholic acid groups and X₃ is a glucose group.