Compositions and methods of vascular injury repair

• US 8,637,005 B2
• Filed: 11/14/2011
• Issued: 01/28/2014
• Est. Priority Date: 11/07/2005
• Status: Active Grant

• Current Assignee: Caladrius Biosciences, Inc. (Lisata Therapeutics, Inc.)
• Original Assignee: Amorcyte, Inc. (Lisata Therapeutics, Inc.)
• Inventors: Pecora, Andrew L., Preti, Robert A.
• Primary Examiner(s): Ford, Allison
• Application Number:
  US13/295,240
• Publication Number:
  US 20120071855A1
• Time in Patent Office: 806 Days
• Field of Search: None
• US Class Current: 424/93.7
• 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: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/10 Antioedematous agents; Diuretics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/06 Antiarrhythmics
• 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 present invention relates to pharmaceutical compositions comprising a chemotactic hematopoietic stem cell product comprising an enriched population of CD34+ cells containing a subpopulation of CD34+/CXCR-4+ cells having CXCR-4-mediated chemotactic activity, methods of preparing these compositions and use of these compositions to treat or repair vascular injury, including infarcted myocardium.

Claims

1. A method of treating or repairing a vascular injury caused by vascular insufficiency in a subject in need thereof, comprising:

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

(a) a therapeutic amount of a sterile chemotactic hematopoietic stem cell product, wherein the therapeutic amount is effective to repair a vascular injury caused by vascular insufficiency, the chemotactic hematopoietic stem cell product comprising a nonexpanded isolated population of autologous mononuclear cells comprising a subpopulation of CD34+ cells, which further contains a subpopulation of potent SDF-1-mobile CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, wherein the therapeutic amount of the chemotactic hematopoietic stem cell product comprises at least 0.5×

10⁶ potent SDF-1 mobile CD34+/CXCR-4+ cells that have CXCR-4 mediated chemotactic activity; and

(b) a stabilizing amount of serum, wherein the stabilizing amount of serum, which ranges from about 0.1% to about 70% (v/v) of the composition, is effective to retain the CXCR-4 mediated chemotactic activity and hematopoietic colony forming activity of the subpopulation of SDF-1 mobile CD34+/CXCR-4+ cells from completion of acquisition of the nonexpanded population of autologous mononuclear cells from the subject to infusion of the chemotactic hematopoietic stem cell product into the subject,

wherein

the pharmaceutical composition is further characterized as having the following properties for at least 24 hours following acquisition of the chemotactic hematopoietic stem cell product, when tested in vitro after passage through the catheter;

(i) the subpopulation of CD34+ cells constitutes from about 1% to about 95% of the isolated nonexpanded population of autologous mononuclear cells,

(ii) the subpopulation of CD34+ cells further contains a subpopulation of potent SDF-1 mobile CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity;

(iii) the subpopulation of CD34+ cells comprises from about 1% to about 95% viable CD34+ cells; and

(iv) the subpopulation of CD34+ cells is able to form hematopoietic colonies in vitro.

2. The method according to claim 1, wherein administering occurs at a plurality of infusion dates.

3. The method according to claim 2, wherein a first infusion date is at least about one day to about 40 years after an occurrence of the vascular injury caused by vascular insufficiency.

4. The method according to claim 3, wherein a second infusion date is at least about one day to about 40 years after an occurrence of the vascular injury caused by vascular insufficiency.

5. The method according to claim 1, wherein the sterile pharmaceutical composition is further characterized as having the following properties for at least 24 hours following acquisition of the chemotactic hematopoietic stem cell product when tested in vitro after passage through the catheter:

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

(b) retains at least 2% of the chemotactic activity of the CXCR-4-mediated chemotactic activity of the subpopulation of potent SDF-1 mobile CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity measured upon acquisition.

6. The method according to claim 1, wherein the sterile pharmaceutical composition is further characterized as having the following properties for at least 48 hours following acquisition of the chemotactic hematopoietic stem cell product when tested in vitro after passage through the catheter:

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

(b) retains at least 2% of the chemotactic activity of the CXCR-4-mediated chemotactic activity of the subpopulation of potent SDF-1 mobile CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity measured upon acquisition.

7. The method according to claim 1, wherein the sterile pharmaceutical composition is further characterized as having the following properties for at least 72 hours following acquisition of the chemotactic hematopoietic stem cell product when tested in vitro after passage through a catheter:

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

(b) retains at least 2% of the chemotactic activity of the CXCR-4-mediated chemotactic activity of the subpopulation of potent SDF-1 mobile CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity measured upon acquisition.

8. The method according to claim 1, wherein parenterally administering comprises delivering the sterile pharmaceutical composition into a coronary blood vessel through the catheter.

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

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

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

12. The method according to claim 1, wherein parenterally administering comprises delivering the sterile pharmaceutical composition into myocardium, an artery, a vein, or a muscle.

13. The method according to claim 1, wherein the isolated, nonexpanded population of autologous mononuclear cells comprising a subpopulation of CD34+ cells, which further contains a subpopulation of potent SDF-1 mobile CD34+/CXCR-4+ cells having CXCR-4-mediated chemotactic activity is purified from peripheral blood collected from the subject.

14. The method according to claim 1, wherein the isolated, nonexpanded population of autologous mononuclear cells comprising a subpopulation of CD34+ cells, which further contains a subpopulation of potent SDF-1 mobile CD34+/CXCR-4+ cells having CXCR-4-mediated chemotactic activity is purified from peripheral blood collected from the subject after mobilizing the isolated, nonexpanded population of autologous mononuclear cells using a hematopoietic stem cell mobilizing agent.

15. The method according to claim 1, wherein the nonexpanded isolated population of autologous mononuclear cells comprising a subpopulation of CD34+ cells, which further contains a subpopulation of potent SDF-1 mobile CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity is purified from cellular components of a bone marrow aspirate harvested from the subject.

16. The method according to claim 1, wherein the sterile pharmaceutical composition further includes at least one compatible active agent.

17. The method according to claim 16, wherein the at least one compatible active agent is selected from the group consisting of a hematopoietic stem cell mobilizing agent, an angiotensin converting enzyme inhibitor, a beta-blocker, a cytokine, a diuretic, an anti-arrhythmic agent, an anti-anginal agent, an anticoagulant, a vasoactive agent, a fibrinolytic agent, a hematopoietic stem cell mobilizing agent, and a hypercholesteromic agent.

18. The method according to claim 1, wherein the vascular insufficiency results from occlusion of a coronary artery.

19. The method according to claim 18, wherein the vascular insufficiency resulting from occlusion of the coronary artery is a microvascular insufficiency.

20. The method according to claim 18, wherein the vascular insufficiency resulting from occlusion of the coronary artery is an ischemia.

21. The method according to claim 18, wherein the vascular insufficiency resulting from occlusion of the coronary artery is a transient vascular insufficiency.

22. The method according to claim 18, wherein the vascular insufficiency resulting from occlusion of the coronary artery is a chronic ischemia.

23. The method according to claim 18, wherein the vascular insufficiency resulting from occlusion of the coronary artery is a myocardial ischemia.

24. The method according to claim 18, wherein the vascular insufficiency resulting from occlusion of the coronary artery produces a myocardial infarction.

25. The method according to claim 1, wherein the potent SDF-1 mobile CD34+/CXCR-4+ cells of the sterile pharmaceutical composition migrate to and repair damage caused by the vascular insufficiency.

26. The method according to claim 25, wherein migration of the potent CD34+/CXCR-4+ cells of the sterile pharmaceutical composition is driven by chemotaxis.

27. The method according to claim 26, wherein the chemotaxis is mediated by at least one of SDF-1 and CXCR-4.

28. The method according to claim 1, wherein sterility of the chemotactic hematopoietic cell product is confirmed by a method comprising the steps:

(a) centrifuging the chemotactic hematopoietic cell product to form (i) a separated cell product comprising a pellet comprising the-isolated population of mononuclear cells comprising a subpopulation of CD34+ cells, which further contains a subpopulation of potent SDF-1 mobile CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity and (ii) a supernatant;

(b) removing the supernatant of the separated cell product without disturbing the cell pellet of the separated cell product;

(c) analyzing the sterility of the supernatant of the separated cell product, thereby determining the sterility of the cell pellet of the separated cell product without depleting the chemotactic hematopoietic cell product.