Compositions and methods for treating progressive myocardial injury due to a vascular insufficiency

Abstract

The described invention provides methods and regimens for treating adverse consequences of a persistent and progressive myocardial injury—due to a vascular insufficiency that occurs early or late in a subject in need thereof, and progressive myocardial injury-preventing compositions that contain a chemotactic hematopoietic stem cell product, and, optionally, an additional active agent.

1 . A method of treating a progressive myocardial injury due to a vascular insufficiency, the method comprising the steps of:
 (a) acquiring a sterile nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity from a subject under sterile conditions;   (b) sterilely enriching the autologous mononuclear cells comprising CD34+ cells, wherein the enriched CD34+ cells which further contain a subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity are a chemotactic hematopoietic stem cell product   (c) administering parenterally through a catheter on a plurality of infusion dates during lifetime of subject a sterile pharmaceutical composition, the sterile pharmaceutical composition comprising:
 (i) a therapeutically effective amount of the sterile chemotactic hematopoietic stem cell product, 
   wherein the therapeutically effective amount of the chemotactic hematopoietic stem cell product comprises at least 10×10 6  CD34+ cells which further contain a subpopulation of at least 0.5×10 6  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 chemotactic hematopoietic stem cell product is further characterized as having the following properties for at least 24 hours when tested in vitro after passage through a catheter:
 (1) retains the CXCR-4-mediated activity of the chemotactic hematopoietic stem cell product; 
 (2) at least 70% of the cells are CD34+ cells; 
 (3) is at least 70% viable; and 
 (4) is able to form hematopoietic colonies in vitro; 
   (d) optionally administering the chemotactic hematopoietic stem cell product at a plurality of infusion dates during the subject's lifetime; and   (e) treating at least one adverse consequence of the progressive vascular insufficiency.   
     
     
         2 . The method according to  claim 1 , step (a) further comprising freezing at least one aliquot of the nonexpanded, isolated population of autologous mononuclear cells containing CD34+ cells, which further contain a subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity at −86° C. and cryostoring the at least one aliquot in the vapor phase of a liquid nitrogen freezer. 
     
     
         3 . The method according to  claim 2 , step (a) further comprising
 (i) thawing the at least one aliquot of the frozen sterile nonexpanded, isolated population of autologous mononuclear cells containing CD34+ cells which further contain a subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity;   (ii) enriching the sterile nonexpanded, isolated population of autologous mononuclear cells for CD34+ cells, which further contain a subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, wherein the sterile nonexpanded, isolated population of autologous mononuclear cells enriched for CD34+ cells which further contain a subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity is a thawed sterile chemotactic hematopoietic stem cell product; and   (iii) administering to the subject on a second infusion date a therapeutically effective amount of the sterile thawed sterile chemotactic hematopoietic stem cell product, comprising
 (a) at least 10×10 6  CD34+ cells, which further contain a subpopulation of at least 0.5×10 6  potent CD34+ cells expressing CXCR-4 and having CXCR-4 mediated chemotactic activity; and 
 (b) a stabilizing amount of serum, wherein the stabilizing amount of serum is greater than 20% (v/v), 
   wherein the sterile thawed chemotactic hematopoietic stem cell product is further characterized as having the following properties for at least 24 hours following thawing of the nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity when tested in vitro after passage through a catheter:   (1) retains the CXCR-4-mediated activity of the subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity;   (2) at least 70% of the cells are CD34+ cells;   (3) is at least 70% viable; and   (4) is able to form hematopoietic colonies in vitro.   
     
     
         4 . The method according to  claim 3 , wherein enriching step (ii) occurs at least 1 day after acquisition of the sterile nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells from the subject. 
     
     
         5 . The method according to  claim 3 , wherein the sterile chemotactic hematopoietic stem cell product is administered parenterally through a catheter to the subject within about 48 hours to about 72 hours of thawing step (i). 
     
     
         6 . The method according to  claim 1 , wherein the nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, is acquired early after an acute myocardial infarction. 
     
     
         7 . The method according to  claim 6 , wherein the nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, is acquired after peak inflammatory cytokine cascade production in an infarcted area. 
     
     
         8 . The method according to  claim 1 , wherein the nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, is acquired late after an acute myocardial infarction. 
     
     
         9 . The method according to  claim 8 , wherein the nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, is acquired at least 15 days after an acute myocardial infarction. 
     
     
         10 . The method according to  claim 3 , wherein the sterile thawed chemotactic hematopoietic stem cell product is further characterized as having the following properties for at least 48 hours following thawing of the nonexpanded, isolated population of autologous mononuclear cells when tested in vitro after passage through a catheter:
 (i) is able to form hematopoietic colonies; and   (ii) retains at least 2% of the CXCR-4-mediated activity of the subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity.   
     
     
         11 . The method according to  claim 3 , wherein the sterile thawed chemotactic hematopoietic stem cell product is further characterized as having the following properties for at least 72 hours following thawing of the nonexpanded, isolated population of autologous mononuclear cells when tested in vitro after passage through a catheter:
 (i) is able to form hematopoietic colonies; and   (ii) retains at least 2% of the CXCR-4-mediated activity of the subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity.   
     
     
         12 . The method according to  claim 1 , wherein the vascular insufficiency is an ischemia. 
     
     
         13 . The method according to  claim 12 , wherein the ischemia is a myocardial ischemia. 
     
     
         14 . The method according to  claim 12 , wherein the ischemia is a transient ischemia. 
     
     
         15 . The method according to  claim 12 , wherein the ischemia is a chronic myocardial ischemia. 
     
     
         16 . The method according to  claim 1 , wherein the vascular insufficiency is a vascular insufficiency after an acute myocardial infarction resulting from underlying disease. 
     
     
         17 . The method according to  claim 16 , wherein the ischemia is a peri-infarct border zone ischemia. 
     
     
         18 . The method according to  claim 16 , wherein a first infusion date comprises a specific time interval defined by a first time and a second time, and wherein the first time is after peak inflammatory cytokine cascade production in an infarcted area and the second time is before myocardial scar formation in the infarcted area. 
     
     
         19 . The method according to  claim 18 , wherein the first time of the first infusion date is at least about 5 days post-infarction. 
     
     
         20 . The method according to  claim 18 , wherein the first time of the first infusion date is about 5 days post-infarction and the second time is about 14 days post-infarction. 
     
     
         21 . The method according to  claim 16 , wherein the method treats cardiomyocyte cell death in the peri-infarct border zone, relative to controls. 
     
     
         22 . The method according to  claim 16 , wherein the method treats hypoperfusion in the peri-infarct border zone, relative to controls. 
     
     
         23 . The method according to  claim 16 , wherein the method treats myocardial hibernation in the peri-infarct border zone, relative to controls. 
     
     
         24 . The method according to  claim 16 , wherein the method decreases infarct area, relative to controls. 
     
     
         25 . The method according to  claim 16 , wherein the method decreases infarct mass, relative to controls. 
     
     
         26 . The method according to  claim 16 , wherein the progressive myocardial injury is a progressive decline in heart muscle function following the acute myocardial infarction. 
     
     
         27 . The method according to  claim 1 , wherein step (e) comprises treating at least one adverse consequence of an acute myocardial infarction selected from premature death, recurrent myocardial infarction, development of congestive heart failure, development of significant arrhythmias, development of acute coronary syndrome, worsening of congestive heart failure, worsening of significant arrhythmias, and worsening of acute coronary syndrome. 
     
     
         28 . The method according to  claim 1 , wherein the progressive myocardial injury is heart failure. 
     
     
         29 . The method according to  claim 1 , wherein the catheter is a flow control catheter. 
     
     
         30 . The method according to  claim 1 , wherein the catheter is a balloon dilatation catheter. 
     
     
         31 . The method according to  claim 1 , wherein the catheter has an internal diameter of at least about 0.36 mm. 
     
     
         32 . The method according to  claim 1 , wherein the administering step (c) is through the catheter into myocardium. 
     
     
         33 . The method according to  claim 1 , wherein the administering step (c) is through the catheter intravascularly. 
     
     
         34 . The method according to  claim 1 , wherein the pharmaceutical composition further includes at least one compatible active agent. 
     
     
         35 . The method according to  claim 34 , wherein the active agent is selected from the group consisting of an angiotensin converting enzyme inhibitor, a beta-blocker, a diuretic, an anti-arrhythmic agent, a hematopoietic stem cell mobilizing agent, a tyrosine kinase receptor agonist, an anti-anginal agent, a vasoactive agent, an anticoagulant agent, a fibrinolytic agent, and a hypercholesterolemic agent. 
     
     
         36 . The method according to  claim 35 , wherein the tyrosine kinase receptor agonist is human neuregulin 1. 
     
     
         37 . A regimen for treating a progressive myocardial injury due to a vascular insufficiency in a revascularized subject, which comprises
 (a) administering parenterally through a catheter on a plurality of infusion dates during lifetime of the subject a sterile pharmaceutical composition comprising a sterile chemotactic hematopoietic stem cell product, wherein the sterile chemotactic hematopoietic stem cell product comprises
 (i) a nonexpanded, isolated population of autologous mononuclear cells enriched for CD34+ cells, which further contain a subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, 
   wherein the therapeutically effective amount of the chemotactic hematopoietic stem cell product comprises at least 10×10 6  CD34+ cells which further contain a subpopulation of at least 0.5×10 6  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 chemotactic hematopoietic stem cell product 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 a catheter:
 (1) retains the CXCR-4-mediated activity of the subpopulation of potent CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity; 
 (2) at least 70% of the cells are CD34+ cells; 
 (3) is at least 70% viable; and 
 (4) is able to form hematopoietic colonies in vitro; and 
   (b) treating at least one adverse consequence of the progressive vascular insufficiency.   
     
     
         38 . The regimen according to  claim 37 , wherein the vascular insufficiency is an ischemia. 
     
     
         39 . The regimen according to  claim 38 , wherein the ischemia is a myocardial ischemia. 
     
     
         40 . The regimen according to  claim 38 , wherein the ischemia is a transient ischemia. 
     
     
         41 . The regimen according to  claim 38 , wherein the ischemia is a chronic myocardial ischemia. 
     
     
         42 . The regimen according to  claim 37 , wherein the vascular insufficiency is a vascular insufficiency after an acute myocardial infarction resulting from underlying disease. 
     
     
         43 . The regimen according to  claim 37 , wherein the nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, is acquired early after occurrence of an acute myocardial infarction. 
     
     
         44 . The regimen according to  claim 43 , wherein the nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, is acquired after peak inflammatory cytokine cascade production in an infarcted area. 
     
     
         45 . The regimen according to  claim 37 , wherein the nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, is acquired late after occurrence of an acute myocardial infarction. 
     
     
         46 . The method according to  claim 45 , wherein the nonexpanded, isolated population of autologous mononuclear cells comprising CD34+ cells, which further contain a subpopulation of CD34+/CXCR-4+ cells that have CXCR-4-mediated chemotactic activity, is acquired at least 15 days after occurrence of an acute myocardial infarction. 
     
     
         47 . The regimen according to  claim 42 , wherein a first infusion date comprises a specific time interval defined by a first time and a second time, and wherein the first time is after peak inflammatory cytokine cascade production in an infarcted area and the second time is before myocardial scar formation in the infarcted area. 
     
     
         48 . The regimen according to  claim 47 , wherein the first time of the first infusion date is at least about 5 days post-infarction. 
     
     
         49 . The regimen according to  claim 47 , wherein the first time of the first infusion date is about 5 days post-infarction and the second time is about 14 days post-infarction. 
     
     
         50 . The regimen according to  claim 42 , wherein the a first infusion date is at least 5 days after occurrence of an acute myocardial infarction. 
     
     
         51 . The regimen according to  claim 42 , wherein a second infusion date is at least 30 days after occurrence of an acute myocardial infarction. 
     
     
         52 . The regimen according to  claim 37 , wherein the ischemia is a peri-infarct border zone ischemia. 
     
     
         53 . The regimen according to  claim 52 , wherein step (b) comprises treating cardiomyocyte cell death in the peri-infarct border zone, relative to controls. 
     
     
         54 . The regimen according to  claim 52 , wherein step (b) comprises treating hypoperfusion in the peri-infarct border zone, relative to controls. 
     
     
         55 . The regimen according to  claim 52 , wherein step (b) comprises treating myocardial hibernation in the peri-infarct border zone, relative to controls. 
     
     
         56 . The regimen according to  claim 52 , wherein step (b) comprises decreasing infarct area, relative to controls. 
     
     
         57 . The regimen according to  claim 52 , wherein step (b) comprises decreasing infarct mass, relative to controls. 
     
     
         58 . The regimen according to  claim 52 , wherein step (b) comprises treating at least one adverse consequence of the acute myocardial infarction selected from premature death, recurrent myocardial infarction, development of congestive heart failure, development of significant arrhythmias, development of acute coronary syndrome, worsening of congestive heart failure, worsening of significant arrhythmias, and worsening of acute coronary syndrome. 
     
     
         59 . The regimen according to  claim 42 , wherein the progressive myocardial injury is a progressive decline in heart muscle function following the acute myocardial infarction. 
     
     
         60 . The regimen according to  claim 37 , wherein the progressive myocardial injury is heart failure. 
     
     
         61 . The regimen according to  claim 37 , wherein the catheter is a flow control catheter. 
     
     
         62 . The regimen according to  claim 37 , wherein the catheter is a balloon dilatation catheter. 
     
     
         63 . The regimen according to  claim 37 , wherein the catheter has an internal diameter of at least about 0.36 mm. 
     
     
         64 . The regimen according to  claim 37 , wherein the composition is administered through the catheter into myocardium. 
     
     
         65 . The regimen according to  claim 37 , wherein the composition is administered through the catheter intravascularly. 
     
     
         66 . The regimen according to  claim 37 , wherein the pharmaceutical composition further includes at least one compatible active agent. 
     
     
         67 . The regimen according to  claim 66 , wherein the active agent is selected from the group consisting of an angiotensin converting enzyme inhibitor, a beta-blocker, a diuretic, an anti-arrhythmic agent, a hematopoietic stem cell mobilizing agent, a tyrosine kinase receptor agonist, an anti-anginal agent, a vasoactive agent, an anticoagulant agent, a fibrinolytic agent, and a hypercholesterolemic agent. 
     
     
         68 . The regimen according to  claim 67 , wherein the tyrosine kinase receptor agonist is human neuregulin 1.