Antisense restenosis composition and method

Abstract

The present invention provides an improved method for reducing the risk or severity of restenosis following cardiac angioplasty. The method includes administering to a target vessel region, a morpholino antisense compound having uncharged phosphorus-containing backbone linkages, and spanning the start codon of a human c-myc mRNA. Also disclosed are novel antisense compounds and compositions, and a method for assaying the effectiveness of antisense delivery and uptake to a target vessel region.

1 . A method for treating a vascular injury site in a human patient by reducing restenosis at the site, said method comprising administering to the patient, by intravascular delivery, an antisense compound having: 
 (i) morpholino subunits linked together by phosphorodiamidate linkages, 2 atoms long, joining the morpholino nitrogen of one subunit to the 5′ exocyclic carbon of an adjacent subunit, and    (ii) a sequence of bases attached to the subunits and containing the sequence identified by SEQ ID NO:1.    
     
     
         2 . The method of  claim 1  wherein the antisense compound has intersubunit linkages of the form:  
       
         
           
           
               
               
           
         
       
       where X is an unsubstituted, monosubstituted or disubstituted nitrogen.  
     
     
         3 . The method of  claim 1 , wherein the antisense compound has from 8 to 40 nucleotides, including a targeting sequence complementary to a region spanning the start codon of human c-myc mRNA, where the nucleotide sequence is SEQ ID NO:1.  
     
     
         4 . The method of  claim 3 , wherein the mode of administration is selected from the group consisting of: 
 (a) contacting the region of the vessel with a reservoir containing the antisense compound, and introducing the compound from the reservoir into the vessel by iontophoresis or electroporation;    (b) injecting the compound from the catheter directly into the region of the vessel, under pressure, through injectors contained on the surface of the catheter balloon, where said injectors are capable of penetrating the tunica media in the vessel;    (c) injecting into or contacting the region of the vessel, microparticles containing the antisense compound in entrapped form;    (d) contacting the region of the vessel with a hydrogel coating contained on the surface of the catheter balloon, and containing the antisense compound is diffusible form; and    (e) contacting the region of the vessel with a stent having an outer surface layer containing the antisense compound in diffusible form.    
     
     
         5 . The method of  claim 4 , wherein the amount of antisense compound administered is between 0.5 and 20 mg.  
     
     
         6 . The method of  claim 4 , in the mode of administration set forth in (a), wherein the antisense compound is contained in a volume between two inflated balloons in the catheter, the compound contains a net charge, and the volume is subjected to pulsed electric fields effective to iontophoretically drive the compound into the vessel region.  
     
     
         7 . The method of  claim 4 , in the mode of administration set forth in (a), wherein the antisense compound is contained in a volume between two inflated balloons in the catheter, and the volume is subjected to pulsed electric fields effective to facilitate compound uptake into vessel-region cells by electroporation.  
     
     
         8 . The method of  claim 4 , in the mode of administration set forth in (b), wherein the catheter balloon has a plurality of outer-facing channels that communicate with a distal-tip reservoir, each channel having one or more injection ports, and said injecting includes forcing a solution or suspension of the antisense compound from said reservoir through said injection ports when the balloon is in an inflated position.  
     
     
         9 . The method of  claim 4 , in the mode of administration set forth in (c), wherein the catheter has a distal end reservoir, the microparticles are contained as a particle suspension in the reservoir, and said injecting includes forcing the suspension out of the catheter through a catheter surface in contact with the vessel region.  
     
     
         10 . The method of  claim 4 , wherein the particles are microbubbles containing the antisense compound in entrapped form.  
     
     
         11 . The method of  claim 4 , in the mode of administration set forth in (e), wherein the stent is biodegradable.  
     
     
         12 . The method of  claim 4 , wherein the compound is derivatized with triethyleneglycol attached to the 5′ end of the compound, and the compound is administered from a solution containing at least 30 mg/ml of the antisense compound.  
     
     
         13 . A morpholino antisense compound having: 
 (i) morpholino subunits linked together by phosphorodiamidate linkages, 2 atoms long, joining the morpholino nitrogen of one subunit to the 5′ exocyclic carbon of an adjacent subunit, and    (ii) a sequence of bases attached to the subunits and containing the sequence identified by SEQ ID NO:1, for the preparation of a pharmaceutical composition for use in inhibiting restenosis in a patient.    
     
     
         14 . A morpholino antisense compound having (i) from 8 to 40 nucleotides, including a targeting nucleic acid sequence complementary to a region that spans the start codon of a human c-myc mRNA gene, and (ii) phosphorous-containing intersubunit linkages.  
     
     
         15 . The compound of  claim 13  having intersubunit linkages of the form:  
       
         
           
           
               
               
           
         
       
       where X is an unsubstituted, monosubstituted or disubstituted nitrogen.  
     
     
         16 . The compound of  claim 13 , wherein the compound is derivatized with triethyleneglycol attached to the 5′ end of the compound.  
     
     
         17 . The compound of  claim 13 , which is entrapped in liposomal or biodegradable microparticles.