Medical devices and methods for delivering compositions to cells

• US 20060018882A1
• Filed: 06/21/2005
• Published: 01/26/2006
• Est. Priority Date: 06/21/2004
• Status: Abandoned Application

• Current Assignee: Medtronic Incorporated (Medtronic Plc)
• Original Assignee: Medtronic Incorporated (Medtronic Plc)
• Inventors: Kaemmerer, William F., Kaytor, Michael D., Burright, Eric N., TenBrock, Erica M., Blum, Janelle L.
• Application Number:
  US11/157,608
• Publication Number:
  US 20060018882A1
• Time in Patent Office: Days
• US Class Current: 424/93.200
• CPC Class Codes: A61K 48/0033 the non-active part being non-polymeric
• CPC Class Codes: A61K 48/0075 characterised by an aspect of the delivery route, e.g. oral, subcutaneous
• CPC Class Codes: A61K 48/0083 characterised by an aspect of the administration regime
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector

Abstract

The present invention provides medical devices and methods for delivering compositions to cells. The compositions include an artificial viral vector, and particularly, an artificial adeno-associated virus vector. Such compositions can be useful for delivering the artificial viral vector across the blood-brain barrier.

Claims

1. A medical system for delivering DNA across a blood-brain barrier, the system comprising:

a neurovascular catheter having a distal end positioned in a blood vessel supplying a patient'"'"'s brain; and

a means for delivering to the catheter a composition comprising;

an artificial adeno-associated virus (AAV) vector comprising DNA encoding a biologically active agent; and

a component to deliver at least the DNA across the blood-brain barrier.

2. The medical system of claim 1 further comprising an implantable pump for delivery of the composition to the patient'"'"'s blood stream.

3. The medical system of claim 1 wherein the artificial AAV vector is for delivery of a single stranded DNA encoding a biologically active agent, the artificial AAV vector comprising the single stranded DNA having AAV-ITRs at the 5-prime and 3-prime ends.

4. The medical system of claim 1 wherein the artificial AAV vector is for delivery of a single stranded DNA encoding a biologically active agent, the artificial AAV vector comprising, in 5-prime to 3-prime order:

a 5-prime AAV-ITR;

the single stranded DNA;

an internal AAV-ITR;

a reverse complement of the single stranded DNA; and

a 3-prime AAV-ITR.

5. The medical system of claim 1 wherein the artificial AAV vector is for delivery of a linear, double stranded DNA encoding a biologically active agent, the artificial AAV vector comprising the linear, double stranded DNA having AAV-ITRs at the 5-prime and 3-prime ends of each strand.

6. The medical system of claim 5 wherein the artificial AAV vector has been thermally treated in at least one heating and cooling cycle.

7. A medical system for delivering DNA across a blood-brain barrier comprising:

a neurovascular catheter having a distal end positioned in a blood vessel supplying a patient'"'"'s brain; and

a means for delivering to the catheter a composition comprising a receptor-specific liposome, wherein the receptor-specific liposome comprises;

a liposome having an exterior surface and an internal compartment;

an artificial adeno-associated virus (AAV) vector located within the internal compartment of the liposome, wherein the artificial AAV vector comprises DNA encoding a biologically active agent;

one or more blood-brain barrier and brain cell membrane targeting agents; and

one or more conjugation agents wherein each targeting agent is connected to the exterior surface of the liposome via at least one of the conjugation agents.

8. The medical system of claim 7 further comprising an implantable pump for delivery of the composition to the patient'"'"'s blood stream.

9. The medical system of claim 7 wherein the exterior surface of the liposome defines a sphere having a diameter of at most 200 nanometers.

10. The medical system of claim 7 wherein at least 5 and at most 1000 blood-brain barrier or brain cell membrane targeting agents are conjugated to the surface of the liposome.

11. The medical system of claim 7 wherein at least 25 and at most 40 blood-brain barrier or brain cell membrane targeting agents are conjugated to the surface of the liposome.

12. The medical system of claim 7 wherein the conjugation agent is selected from the group consisting of polyethylene glycol, sphingomyelin, biotin, streptavidin, organic polymers, and combinations thereof.

13. The medical system of claim 7 wherein the molecular weight of the conjugation agent is at least 1000 Daltons and at most 50,000 Daltons.

14. The medical system of claim 7 wherein the artificial AAV vector comprises a sequence selected from the group consisting of SEQ ID NOs:

8-11.

15. The medical system of claim 7 wherein the DNA encoding the biologically active agent comprises a sequence selected from the group consisting of SEQ ID NOs:

1-7.

16. The medical system of claim 7 wherein the DNA encodes a short hairpin RNA.

17. The medical system of claim 7 wherein the DNA encodes a protein.

18. A method for delivering DNA across a blood-brain barrier for expression in the brain, the method comprising administering to a patient a composition comprising a receptor-specific liposome, wherein the receptor-specific liposome comprises:

a liposome having an exterior surface and an internal compartment;

an artificial adeno-associated virus (AAV) vector located within the internal compartment of the liposome, wherein the artificial AAV vector comprises DNA encoding a biologically active agent;

one or more blood-brain barrier and brain cell membrane targeting agents; and

one or more conjugation agents wherein each targeting agent is connected to the exterior surface of the liposome via at least one of the conjugation agents.

19. The method of claim 18 wherein the composition is administered intravenously or intra-arterially.

20. The method of claim 18 wherein the exterior surface of the liposome defines a sphere having a diameter of at most 200 nanometers.

21. The method of claim 18 wherein at least 5 and at most 1000 blood-brain barrier or brain cell membrane targeting agents are conjugated to the surface of the liposome.

22. The method of claim 18 wherein at least 25 and at most 40 blood-brain barrier or brain cell membrane targeting agents are conjugated to the surface of the liposome.

23. The method of claim 18 wherein the conjugation agent is selected from the group consisting of polyethylene glycol, sphingomyelin, biotin, streptavidin, organic polymers, and combinations thereof.

24. The method of claim 18 wherein the molecular weight of the conjugation agent is at least 1000 Daltons and at most 50,000 Daltons.

25. A method for delivering DNA to a cell, the method comprising administering to a patient a composition comprising a receptor-specific nanocontainer, wherein the receptor-specific nanocontainer comprises:

a nanocontainer having an exterior surface and an internal compartment;

an artificial adeno-associated virus (AAV) vector located within the internal compartment of the nanocontainer, wherein the artificial AAV vector comprises DNA encoding a biologically active agent;

one or more receptor specific targeting agents that target the receptor located on the cell; and

one or more conjugation agents wherein each targeting agent is connected to the exterior surface of the nanocontainer via at least one of the conjugation agents.

26. The method of claim 25 wherein the composition is administered intravenously or intra-arterially.

27. The method of claim 25 wherein the exterior surface of the nanocontainer defines a sphere having a diameter of at most 200 nanometers.

28. The method of claim 25 wherein the cells are selected from the group consisting of brain cells, liver cells, lung cells, and spleen cells.

29. The method of claim 25 wherein the artificial AAV vector is for delivery of a single stranded DNA encoding a biologically active agent, the artificial AAV vector comprising the single stranded DNA having AAV-ITRs at the 5-prime and 3-prime ends.

30. The method of claim 25 wherein the artificial AAV vector is for delivery of a single stranded DNA encoding a biologically active agent, the artificial AAV vector comprising, in 5-prime to 3-prime order:

a 5-prime AAV-ITR;

the single stranded DNA;

an internal AAV-ITR;

a reverse complement of the single stranded DNA; and

a 3-prime AAV-ITR.

31. The method of claim 25 wherein the artificial AAV vector is for delivery of a linear, double stranded DNA encoding a biologically active agent, the artificial AAV vector comprising the linear, double stranded DNA having AAV-ITRs at the 5-prime and 3-prime ends of each strand.

32. The method of claim 31 wherein the artificial AAV vector has been thermally treated in at least one heating and cooling cycle.

33. The method of claim 25 wherein the DNA encodes a short hairpin RNA.

34. The method of claim 33 wherein the short hairpin RNA is expressed in the cell.

35. The method of claim 25 wherein the DNA encodes a protein.

36. The method of claim 35 wherein the protein is expressed in the cell.

37. A method for delivering DNA across a blood-brain barrier for expression in the brain, the method comprising administering to a patient a composition comprising:

an artificial adeno-associated virus (AAV) vector comprising DNA encoding a biologically active agent; and

a component to deliver at least the DNA across the blood-brain barrier.

38. The method of claim 37 wherein the composition is administered intravenously or intra-arterially.

39. The method of claim 37 wherein the artificial AAV vector comprises a sequence selected from the group consisting of SEQ ID NOs:

8-11.

40. The method of claim 37 wherein the DNA encoding the biologically active agent comprises a sequence selected from the group consisting of SEQ ID NOs:

1-7.

41. The method of claim 37 wherein the DNA encodes a short hairpin RNA.

42. The method of claim 41 wherein the short hairpin RNA is expressed in the brain.

43. The method of claim 37 wherein the DNA encodes a protein.

44. The method of claim 43 wherein the protein is expressed in the brain.

45. A method of treating a neurodegenerative disorder caused by a pathogenic protein, the method comprising:

providing a neurovascular catheter having a distal end positioned in a blood vessel supplying a patient'"'"'s brain; and

delivering to the catheter a composition comprising;

an artificial adeno-associated virus (AAV) vector comprising DNA encoding a biologically active agent; and

a component to deliver at least the DNA across the blood-brain barrier.

46. The method of claim 45 wherein the DNA encodes a short hairpin RNA.

47. The method of claim 46 wherein the short hairpin RNA is expressed in the brain.

48. A method of treating a neurodegenerative disorder caused by a pathogenic protein, the method comprising:

providing a neurovascular catheter having a distal end positioned in a blood vessel supplying a patient'"'"'s brain; and

delivering to the catheter a composition comprising a receptor-specific liposome and a pharmaceutically acceptable carrier for the receptor-specific liposome, wherein the receptor-specific liposome comprises;

a liposome having an exterior surface and an internal compartment;

an artificial adeno-associated virus (AAV) vector located within the internal compartment of the liposome, wherein the artificial AAV vector comprises DNA encoding a biologically active agent;

one or more blood-brain barrier and brain cell membrane targeting agents; and

one or more conjugation agents wherein each targeting agent is connected to the exterior surface of the liposome via at least one of the conjugation agents.

49. The method of claim 48 wherein the DNA encodes a short hairpin RNA.

50. The method of claim 49 wherein the short hairpin RNA is expressed in the brain.

51. A method of treating a neurological disease caused by the absence of a protein, the method comprising:

providing a neurovascular catheter having a distal end positioned in a blood vessel supplying a patient'"'"'s brain; and

delivering to the catheter a composition comprising;

an artificial adeno-associated virus (AAV) vector comprising DNA encoding a biologically active agent; and

a component to deliver at least the DNA across the blood-brain barrier.

52. The method of claim 51 wherein the DNA encodes a protein.

53. The method of claim 52 wherein the protein is expressed in the brain.

54. The method of claim 51 wherein the neurological disease is an inborn error of metabolism.

55. A method of treating a neurological disease caused by the absence of a protein, the method comprising:

providing a neurovascular catheter having a distal end positioned in a blood vessel supplying a patient'"'"'s brain; and

delivering to the catheter a composition comprising a receptor-specific liposome and a pharmaceutically acceptable carrier for the receptor-specific liposome, wherein the receptor-specific liposome comprises;

a liposome having an exterior surface and an internal compartment;

an artificial adeno-associated virus (AAV) vector located within the internal compartment of the liposome, wherein the artificial AAV vector comprises DNA encoding a biologically active agent;

one or more blood-brain barrier and brain cell membrane targeting agents; and

one or more conjugation agents wherein each targeting agent is connected to the exterior surface of the liposome via at least one of the conjugation agents.

56. The method of claim 55 wherein the DNA encodes a protein.

57. The method of claim 56 wherein the protein is expressed in the brain.

58. The method of claim 55 wherein the neurological disease is an inborn error of metabolism.

59. A composition for delivering DNA across a blood-brain barrier for expression in the brain, the composition comprising a receptor-specific liposome, wherein the receptor-specific liposome comprises:

a liposome having an exterior surface and an internal compartment;

an artificial adeno-associated virus (AAV) vector located within the internal compartment of the liposome, wherein the artificial AAV vector comprises DNA encoding a biologically active agent;

one or more blood-brain barrier and brain cell membrane targeting agents; and

one or more conjugation agents wherein each targeting agent is connected to the exterior surface of the liposome via at least one of the conjugation agents.

60. The composition of claim 59 wherein the exterior surface of the liposome defines a sphere having a diameter of at most 200 nanometers.

61. The composition of claim 59 wherein at least 5 and at most 1000 blood-brain barrier or brain cell membrane targeting agents are conjugated to the surface of the liposome.

62. The composition of claim 59 wherein at least 25 and at most 40 blood-brain barrier or brain cell membrane targeting agents are conjugated to the surface of the liposome.

63. The composition of claim 59 wherein the conjugation agent is selected from the group consisting of polyethylene glycol, sphingomyelin, biotin, streptavidin, organic polymers, and combinations thereof.

64. The composition of claim 59 wherein the molecular weight of the conjugation agent is at least 1000 Daltons and at most 50,000 Daltons.

65. A composition for delivering DNA to a cell, the composition comprising a receptor-specific nanocontainer, wherein the receptor-specific nanocontainer comprises:

a nanocontainer having an exterior surface and an internal compartment;

an artificial adeno-associated virus (AAV) vector located within the internal compartment of the nanocontainer, wherein the artificial AAV vector comprises DNA encoding a biologically active agent;

one or more receptor specific targeting agents that target the receptor located on the cell; and

one or more conjugation agents wherein each targeting agent is connected to the exterior surface of the nanocontainer via at least one of the conjugation agents.

66. The composition of claim 65 wherein the exterior surface of the nanocontainer defines a sphere having a diameter of at most 200 nanometers.

67. The composition of claim 65 wherein the artificial AAV vector is for delivery of a single stranded DNA encoding a biologically active agent, the artificial AAV vector comprising the single stranded DNA having AAV-ITRs at the 5-prime and 3-prime ends.

68. The composition of claim 65 wherein the artificial AAV vector is for delivery of a single stranded DNA encoding a biologically active agent, the artificial AAV vector comprising, in 5-prime to 3-prime order:

a 5-prime AAV-ITR;

the single stranded DNA;

an internal AAV-ITR;

a reverse complement of the single stranded DNA; and

a 3-prime AAV-ITR.

69. The composition of claim 65 wherein the artificial AAV vector is for delivery of a linear, double stranded DNA encoding a biologically active agent, the artificial AAV vector comprising the linear, double stranded DNA having AAV-ITRs at the 5-prime and 3-prime ends of each strand.

70. The composition of claim 69 wherein the artificial AAV vector has been thermally treated in at least one heating and cooling cycle.

71. A composition for delivering DNA across a blood-brain barrier for expression in the brain, the composition comprising:

an artificial adeno-associated virus (AAV) vector comprising DNA encoding a biologically active agent; and

a component to deliver at least the DNA across the blood-brain barrier.

72. The composition of claim 71 wherein the artificial AAV vector comprises a sequence selected from the group consisting of SEQ ID NOs:

8-11.

73. The composition of claim 71 wherein the DNA encoding the biologically active agent comprises a sequence selected from the group consisting of SEQ ID NOs:

1-7.

74. The composition of claim 71 wherein the DNA encodes a short hairpin RNA.

75. The composition of claim 71 wherein the DNA encodes a protein.

76. An artificial adeno-associated virus (AAV) vector comprising, in 5-prime to 3-prime order:

a 5-prime AAV-ITR;

a single stranded DNA encoding a biologically active agent;

an internal AAV-ITR;

a reverse complement of the single stranded DNA encoding the biologically active agent; and

a 3-prime AAV-ITR.

77. The vector of claim 76 wherein the artificial AAV vector comprises a sequence selected from the group consisting of SEQ ID NOs:

10-11.

78. The vector of claim 76 wherein the DNA encoding the biologically active agent comprises a sequence selected from the group consisting of SEQ ID NOs:

1-7.

79. The vector of claim 76 wherein the DNA encodes a short hairpin RNA.

80. The vector of claim 76 wherein the DNA encodes a protein.

81. An artificial adeno-associated virus (AAV) vector for delivery of a linear, double stranded DNA encoding a biologically active agent, the artificial AAV vector comprising the linear, double stranded DNA having AAV-ITRs at the 5-prime and 3-prime ends of each strand.

82. The vector of claim 81 wherein the artificial AAV vector has been thermally treated in at least one heating and cooling cycle.

83. The vector of claim 81 wherein the DNA encoding the biologically active agent comprises a sequence selected from the group consisting of SEQ ID NOs:

1-7.

84. The vector of claim 81 wherein the DNA encodes a short hairpin RNA.

85. The vector of claim 81 wherein the DNA encodes a protein.

86. A method of making an artificial adeno-associated virus (AAV) vector comprising:

assembling in a DNA plasmid through a DNA cloning method, in 5-prime to 3-prime order, a 5-prime AAV inverted terminal repeat (AAV-ITR), a DNA encoding a biologically active agent, and a 3-prime AAV-ITR;

generating reaction products comprising a single stranded RNA transcript of a single stranded DNA from the DNA plasmid through an ill vitro transcription method;

generating a single stranded DNA from the RNA transcript in the reaction products by reverse transcription through a reverse transcription method; and

removing the RNA transcript from the reaction products by digestion of the RNA using an RNase enzyme.

87. The method of claim 86 further comprising purifying the single stranded DNA from the reaction products by a DNA purification method selected from the group consisting of gel purification, column affinity methods, and combinations thereof.

88. A method of making an artificial adeno-associated virus (AAV) vector comprising:

assembling in a circular DNA plasmid through a DNA cloning method, in 5-prime to 3-prime order, a 5-prime AAV inverted terminal repeat (AAV-ITR), a DNA encoding a biologically active agent, and a 3-prime AAV-ITR;

linearizing the circular plasmid by digesting the plasmid with a restriction enzyme that cuts the DNA at a single, known location in the plasmid sequence just 5-prime to the 5-prime AAV-ITR;

chemically conjugating an affinity tag to the 5-prime ends of each strand of the linearized plasmid;

cutting the DNA sequence with a restriction enzyme that cuts the DNA at a different single, known location in the plasmid sequence just 3-prime to the 3-prime AAV-ITR, such that the restriction digest results in two linear double stranded DNA segments of different sizes;

separating the populations of DNA segments by size using a size separation method and recovering a double stranded DNA;

melting the double stranded DNA to separate its two complementary strands into two single strands, and passing the mixture through an affinity column for the affinity tag such that the strand which was tagged is captured on the column while the non-tagged single strand flows through as the final product.

89. The method of claim 88 wherein the affinity tag comprises a biotin molecule and the affinity column comprises a streptavidin affinity column.

90. The method of claim 88 wherein the size separation method is selected from the group consisting of column filtration, gel electrophoresis, and combinations thereof.

91. A method of making an artificial adeno-associated virus (AAV) vector comprising:

assembling in a circular DNA plasmid through a DNA cloning method, in 5-prime to 3-prime order, a 5-prime AAV inverted terminal repeat (AAV-ITR), a DNA encoding a biologically active agent, and a 3-prime AAV-ITR;

linearizing the circular plasmid by digesting the plasmid with a restriction enzyme that cuts the DNA at a single, known location in the plasmid sequence just 5-prime to the 5-prime AAV-ITR;

chemically conjugating an affinity tag to the 5-prime ends of each strand of the linearized plasmid;

cutting the DNA sequence with a restriction enzyme that cuts the DNA at a different single, known location in the plasmid sequence just 3-prime to the 3-prime AAV-ITR, such that the restriction digest results in two linear double stranded DNA segments of different sizes;

separating the populations of DNA segments by size using a size separation method and recovering a double stranded DNA.

92. The method of claim 91 wherein the size separation method is selected from the group consisting of column filtration, gel electrophoresis, and combinations thereof.

93. The method of claim 91 further comprises thermally treating the recovered double stranded DNA in at least one heating and cooling cycle.

94. A method of making a self complementary, artificial adeno-associated virus (AAV) vector comprising:

assembling in a DNA plasmid through a DNA cloning method, in 5-prime to 3-prime order, a 5-prime AAV inverted terminal repeat (AAV-ITR), a DNA encoding a biologically active agent, an internal AAV-ITR, a reverse complement of the a DNA encoding the biologically active agent, and a 3-prime AAV-ITR;

linearizing the circular plasmid by digesting the plasmid with restriction enzymes that cut out a DNA sequence comprising in 5-prime to 3-prime order, a 5-prime AAV inverted terminal repeat (AAV-ITR), a DNA encoding a biologically active agent, an internal AAV-ITR, a reverse complement of the a DNA encoding the biologically active agent, and a 3-prime AAV-ITR;

recovering a double stranded DNA by using a size separation method;

melting the double stranded DNA to separate its two complementary strands into two single strands; and

lowering the temperature of the melted DNA to allow the single strands to self-anneal into a hairpin form.

95. The method of claim 94 wherein the size separation method is selected from the group consisting of column filtration, gel electrophoresis, and combinations thereof.