INFARCT AREA PERFUSION-IMPROVING COMPOSITIONS AND METHODS OF VASCULAR INJURY REPAIR

• US 20100143317A1
• Filed: 12/02/2009
• Published: 06/10/2010
• Est. Priority Date: 10/24/2006
• Status: Active Grant

• Current Assignee: Caladrius Biosciences, Inc. (Lisata Therapeutics, Inc.)
• Original Assignee: Amorcyte, LLC (Lisata Therapeutics, Inc.)
• Inventors: Pecora, Andrew, Preti, Robert
• Granted Patent: US 9,034,316 B2
• Time in Patent Office: Days
• US Class Current: 424/93.720
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/155 Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
• CPC Class Codes: A61K 31/22 of acyclic acids, e.g. pravastatin
• CPC Class Codes: A61K 31/4439 containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole nicotine A61K31/465
• CPC Class Codes: A61K 31/5375 1,4-Oxazines, e.g. morpholine
• CPC Class Codes: A61K 35/16 Blood plasma; Blood serum umbilical cord blood A61K35/51
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 38/085 Angiotensins
• CPC Class Codes: A61K 38/10 Peptides having 12 to 20 amino acids A61K38/043 - A61K38/046 take precedence
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

The described invention provides pharmaceutical compositions for treating an infarct area injury and methods of treating or repairing the infarct area injury in a revascularized subject in the aftermath of an acute myocardial infarction resulting from a natural disease process by administering to the subject parenterally through a catheter a sterile pharmaceutical composition containing a therapeutically effective amount of a nonexpanded sterile isolated chemotactic hematopoietic stem cell product as a first therapeutic agent and optionally a therapeutically effective amount of at least one compatible second therapeutic agent. The infarct area-improving amount of the sterile isolated chemotactic hematopoietic stem cell product comprises an enriched population of isolated autologous CD34+ cells containing a subpopulation of potent cells expressing CXCR-4 and having CXCR-4-mediated chemotactic activity such that the enriched population of isolated autologous CD34+ hematopoietic stem cells provides at least 0.5×10⁶ potent CD34⁺ cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity.

Claims

1. A method of treating or repairing an infarct area injury in a revascularized subject following an acute myocardial infarction resulting from a natural disease process, the method comprising the steps:

(a) administering to the subject parenterally through a catheter a sterile pharmaceutical composition comprising;

(i) an infarct area perfusion-improving amount of a nonexpanded sterile isolated chemotactic hematopoietic stem cell product as a first therapeutic agent,

wherein the infarct area perfusion-improving amount of the chemotactic hematopoietic stem cell product comprises an enriched population of isolated autologous CD34+ hematopoietic stem cells containing a subpopulation of potent CD34+ cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity such that the enriched population of isolated autologous CD34+ hematopoietic stem cells provides at least 0.5×

10⁶ potent CD34⁺ cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity;

(ii) a stabilizing amount of serum, wherein the stabilizing amount of serum is greater than 20% (v/v), and

(iii) optionally a therapeutically effective amount of at least one compatible second therapeutic agent; and

(b) improving perfusion in at least one infarct area, relative to controls,

wherein at least 70% of cells in the enriched population of isolated CD34+ cells containing the subpopulation of potent cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity when passed through the catheter and when tested in vitro are CD34+ cells, and

wherein the enriched population of isolated CD34+ cells containing a subpopulation of potent cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity when passed through the catheter and tested in vitro;

(1) retains the CXCR-4-mediated chemotactic activity;

(2) is at least about 70% viable; and

(3) is able to form hematopoietic colonies in vitro

for at least about 24 hours following acquisition from the subject of the enriched population of CD34+ cells containing the subpopulation of potent cells that express CXCR-4; and

wherein administering step (a) occurs at one or more infusion dates, and

wherein a first infusion date comprises a specific time interval defined by a first time and a second time, wherein the first time is after peak inflammatory cytokine cascade production in the infarcted area and the second time is before myocardial scar formation in the infarcted area.

2. The method according to claim 1, wherein the infarct area perfusion-improving amount of the chemotactic hematopoietic stem cell product comprises an enriched population of at least 10×

10⁶ isolated autologous CD34+ hematopoietic stem cells containing a subpopulation of 0.5×

10⁶ potent CD34+ cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity.

3. The method according to claim 1, wherein the infarct area injury comprises apoptotic cardiomyocyte loss in the infarct area.

4. The method according to claim 1, wherein the infarct area injury comprises adverse ventricular remodeling after an acute myocardial infarction, when compared to controls.

5. The method according to claim 1, wherein the infarct area injury comprises a progressive decline in heart muscle function following the acute myocardial infarction.

6. The method according to claim 1, wherein the infarct area injury comprises hypoperfusion of at least one ischemic peri-infarct zone of myocardial tissue.

7. The method according to claim 1, wherein the infarct area injury comprises myocardial hibernation in the peri-infarct border zone.

8. The method according to claim 1, further comprising the step:

administering at a second infusion date a frozen and thawed second aliquot of the second sterile pharmaceutical composition, the frozen and thawed second aliquot comprising

(i) a frozen and thawed enriched population of isolated autologous CD34⁺ hematopoietic stem cells containing a subpopulation of at least 0.5×

10⁶ potent CD34⁺ cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity; and

(ii) a stabilizing amount of serum, wherein the stabilizing amount of serum is greater than 20% (v/v),

wherein

the frozen and thawed enriched population of isolated CD34+ cells containing the subpopulation of potent cells expressing CXCR-4 and having CXCR-4-mediated chemotactic activity of the second aliquot, when passed through the catheter and tested in vitro;

(1) retains the CXCR-4-mediated activity;

(2) contains at least 70% CD34+ cells;

(3) is at least 70% viable; and

(4) is able to form hematopoietic colonies in vitro

for at least about 24 hours following thawing of the second aliquot.

9. The method according to claim 8, wherein the method further comprises the step:

optionally administering, at a third infusion date, a frozen and thawed third aliquot of the sterile pharmaceutical composition, the third aliquot comprising a frozen and thawed enriched population of isolated autologous CD34⁺ hematopoietic stem cells containing a subpopulation of at least 0.5×

10⁶ potent CD34⁺ cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity;

(ii) a stabilizing amount of serum, wherein the stabilizing amount of serum is greater than 20% (v/v),

wherein

the frozen and thawed enriched population of isolated CD34+ cells containing the subpopulation of potent cells expressing CXCR-4 and having CXCR-4-mediated chemotactic activity of the third aliquot, when passed through the catheter and tested in vitro;

(1) retains the CXCR-4-mediated activity;

(2) contains at least 70% CD34+ cells;

(3) is at least 70% viable; and

(4) is able to form hematopoietic colonies in vitro,

for at least about 24 hours following thawing of the third aliquot.

10. The method according to claim 8, wherein the second infusion date is about 30 days after the first infusion date.

11. The method according to claim 9, wherein the third infusion date is about 60 days after the first infusion date.

12. The method according to claim 1, wherein the enriched population of CD34⁺ cells containing a subpopulation of potent cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity

(a) is capable of forming hematopoietic colonies in vitro;

and

(b) retains at least 2% of the CXCR-4-mediated chemotactic activity

for at least 48 hours following acquisition of the of the enriched population of CD34+ cells containing a subpopulation of potent cells that express CXCR-4 in (a).

13. The method according to claim 1, wherein the enriched population of CD34⁺ cells containing a subpopulation of potent cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity

(a) is capable of forming hematopoietic colonies in vitro;

and

(b) retains at least 2% of the CXCR-4-mediated chemotactic activity

for at least 72 hours following acquisition of the enriched population of CD34+ cells containing a subpopulation of potent cells that express CXCR-4 in (a).

14. The method according to claim 1, wherein the subpopulation of potent cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity retains at least 2% of the CXCR-4-mediated chemotactic activity for at least 24 hours following acquisition from the subject of the enriched population of CD34+ cells containing a subpopulation of potent cells that express CXCR-4 in (a).

15. The method according to claim 1, the method further comprising the step of delivering the composition intravascularly to an infarct related artery.

16. The method according to claim 1, further comprising the step of delivering the composition through the catheter into myocardium.

17. The method according to claim 1, wherein the catheter is a flow control catheter.

18. The method according to claim 1, wherein the catheter is a balloon dilatation catheter.

19. The method according to claim 1, wherein the catheter has an internal diameter of at least about 0.36 mm.

20. The method according to claim 1, wherein the first time of the specific time interval of the first infusion date is at least about 5 days post-infarction.

21. The method according to claim 1, wherein the second time of the specific time interval of the first infusion date is less than about 14 days post-infarction.

22. The method according to claim 1, wherein the first time of the specific time interval of the first infusion date is at least about 5 days post-infarction and the second time of the specific time interval of the first infusion date is less than about 14 days post-infarction.

23. The method according to claim 1, wherein the optional second therapeutic agent is at least one compatible agent that promotes cardiomyocyte growth.

24. The method according to claim 1, wherein the optional second therapeutic agent is selected from the group consisting of an angiotensin converting enzyme inhibitor, a beta-blocker, a diuretic, an anti-arrhythmic agent, an anti-anginal agent, a tyrosine kinase receptor agonist, a vasoactive agent, an anticoagulant agent, a fibrinolytic agent, and a hypercholesterolemic agent.

25. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises the tyrosine kinase receptor agonist neuregulin 1.

26. The method according to claim 23, wherein the method reduces the infarct area injury more than components (i) plus (ii) or component (iii) alone.

27. The method according to claim 1, wherein the method improves microvascular blood flow in the infarct area, when compared to controls.

28. The method according to claim 1, wherein the method decreases area of the infarct injury, when compared to controls.

29. The method according to claim 1, wherein the method decreases infarct mass, when compared to controls.

30. The method according to claim 1, wherein the method increases perfusion of at least one ischemic peri-infarct zone of myocardial tissue, when compared to controls.

31. The method according to claim 1, wherein the method increases perfusion to hibernating myocardium in at least one peri-infarct zone of myocardial tissue, when compared to controls.

32. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises a vascular endothelial growth factor selected from the group consisting of VEGF-A, VEGF-B, VEGF-C, and VEGF-D.

33. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises placental growth factor.

34. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyoctyte growth comprises a catecholamine.

35. The method according to claim 34, wherein the catecholamine is norepinephrine.

36. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises endothelin-1.

37. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises a prostaglandin F_2α

.

38. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises angiotensin II.

39. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises a phorbol ester.

40. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises neuropeptide Y.

41. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises active transforming growth factor β

1.

42. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises G_q protein.

43. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises diacyl glyercol.

44. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises salusin-α

.

45. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises salusin-β

.

46. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises insulin-like growth factor-1.

47. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises myostatin.

48. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises granulocyte colony-stimulating factor.

49. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises macrophage colony-stimulating factor.

50. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises a thiazolidinedione.

51. The method according to claim 50, wherein the thiazolidinedione is rosiglitazone.

52. The method according to claim 23, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises tumor necrosis factor-like weak inducer of apoptosis.

53. A pharmaceutical composition for treating an infarct area injury of a revascularized subject following an acute myocardial infarction resulting from a natural disease process, comprising:

(a) an infarct-injury improving amount of a sterile isolated chemotactic hematopoietic stem cell product,

wherein the infarct area-improving amount of the sterile isolated chemotactic hematopoietic stem cell product comprises an enriched population of isolated autologous CD34+ cells containing a subpopulation of potent cells expressing CXCR-4 and having CXCR-4-mediated chemotactic activity such that the enriched population of isolated autologous CD34+ hematopoietic stem cells provides at least 0.5×

10⁶ potent CD34⁺ cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity;

(b) a stabilizing amount of serum, wherein the stabilizing amount of serum is greater than 20% (v/v), and

(c) a therapeutically effective amount of at least one compatible agent that promotes cardiomyocyte growth,

wherein the composition is administered to the subject parenterally through a catheter;

wherein at least 70% of cells in the enriched population of isolated CD34+ cells containing the subpopulation of potent cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity when passed through the catheter and when tested in vitro are CD34+ cells, and

wherein the enriched population of isolated CD34+ cells containing a subpopulation of potent cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity when passed through the catheter and tested in vitro;

(1) retains the CXCR-4-mediated chemotactic activity;

(2) is at least about 70% viable; and

(3) is able to form hematopoietic colonies in vitro for at least about 24 hours following acquisition from the subject of the enriched population of CD34+ cells containing the subpopulation of potent cells that express CXCR-4.

54. The composition according to claim 53, wherein the infarct area perfusion-improving amount of the chemotactic hematopoietic stem cell product comprises an enriched population of at least 10×

10⁶ isolated autologous CD34+ hematopoietic stem cells containing a subpopulation of 0.5×

10⁶ potent CD34+ cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity.

55. The composition according to claim 53, wherein the composition reduces the infarct area injury more than components (i) plus (ii) or component (iii) alone.

56. The pharmaceutical composition according to claim 53, wherein the infarct area injury comprises apoptotic cardiomyocyte loss in the infarct area.

57. The pharmaceutical composition according to claim 53, wherein the infarct area injury comprises adverse ventricular remodeling after an acute myocardial infarction, when compared to controls.

58. The pharmaceutical composition according to claim 53, wherein the infarct area injury comprises a progressive decline in heart muscle function following the acute myocardial infarction.

59. The pharmaceutical composition according to claim 53, wherein the infarct area injury comprises hypoperfusion of at least one ischemic peri-infarct zone of myocardial tissue.

60. The pharmaceutical composition according to claim 53, wherein the infarct area injury comprises myocardial hibernation in the peri-infarct border zone.

61. The pharmaceutical composition according to claim 53, wherein the enriched population of isolated CD34+ cells containing a subpopulation of potent cells expressing CXCR-4 and having CXCR-4-mediated chemotactic activity is purified from cellular components of a bone marrow aspirate acquired from the subject.

62. The pharmaceutical composition according to claim 53, wherein the enriched population of isolated CD34+ cells containing a subpopulation of potent cells expressing CXCR-4 and having CXCR-4-mediated chemotactic activity is purified from peripheral blood.

63. The pharmaceutical composition according to claim 53, wherein the enriched population of CD34+ cells containing a subpopulation of potent cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity

(a) is capable of forming hematopoietic colonies in vitro;

and

(b) retains at least 2% of the CXCR-4-mediated chemotactic activity

for at least about 48 hours following acquisition from the subject of the enriched population of CD34+ cells containing the subpopulation of potent cells that express CXCR-4 in (a).

64. The composition according to claim 53, wherein the enriched population of CD34+ cells containing a subpopulation of potent cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity

(a) is capable of forming hematopoietic colonies in vitro;

and

(b) retains at least 2% of the CXCR-4-mediated chemotactic activity

for at least about 72 hours following acquisition from the subject of the enriched population of CD34+ cells containing the subpopulation of potent cells that express CXCR-4 in (a).

65. The composition according to claim 53, wherein the enriched population of CD34+ cells containing a subpopulation of potent cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity retains at least 2% of the CXCR-4-mediated chemotactic activity for at least about 24 hours following acquisition from the subject of the enriched population of CD34+ cells containing the subpopulation of potent cells that express CXCR-4 in (a).

66. The composition according to claim 53, wherein the composition is administered through the catheter intravascularly to an infarct-related artery.

67. The composition according to claim 53 wherein the composition is administered through the catheter into myocardium.

68. The composition according to claim 53, wherein the composition improves microvascular blood flow in the infarct area, when compared to controls.

69. The composition according to claim 53, wherein the composition increases perfusion of at least one ischemic peri-infarct zone of myocardial tissue, when compared to controls.

70. The composition according to claim 53, wherein the composition increases perfusion to hibernating myocardium in at least one peri-infarct zone of myocardial tissue, when compared to controls.

71. The composition according to claim 53, wherein the composition decreases area of infarct area injury, when compared to controls.

72. The composition according to claim 53, wherein the composition decreases infarct mass, when compared to controls.

73. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth is selected from the group consisting of an angiotensin converting enzyme inhibitor, a beta-blocker, a diuretic, an anti-arrhythmic agent, an anti-anginal agent, a tyrosine kinase receptor agonist, a vasoactive agent, an anticoagulant agent, a fibrinolytic agent, and a hypercholesterolemic agent.

74. The composition according to claim 73, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises the tyrosine kinase receptor agonist neuregulin 1.

75. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises an vascular endothelial growth factor selected from the group consisting of VEGF-A. VEGF-B, VEGF-C, and VEGF-D.

76. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises placental growth factor.

77. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyoctyte growth comprises a catecholamine.

78. The composition according to claim 77, wherein the catecholamine is norepinephrine.

79. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises endothelin-1.

80. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises a prostaglandin F_2α

.

81. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises angiotensin II.

82. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises a phorbol ester.

83. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises neuropeptide Y.

84. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises active transforming growth factor β

1.

85. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises G_q protein.

86. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises diacyl glyercol.

87. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises salusin-α

.

88. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises salusin-β

.

89. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises insulin-like growth factor-1.

90. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises myostatin.

91. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises granulocyte colony-stimulating factor.

92. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises macrophage colony-stimulating factor.

93. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises a thiazolidinedione.

94. The composition according to claim 93, wherein the thiazolidinedione is rosiglitazone.

95. The composition according to claim 53, wherein the at least one compatible agent that promotes cardiomyocyte growth comprises tumor necrosis factor-like weak inducer of apoptosis.