Gene therapy for treating familial hypercholesterolemia

Abstract

Compositions and regimens useful in reducing one or more of: LDL-cholesterol, total cholesterol, and/or fasting triglycerides in a subject, and/or modifying fractional catabolic rate (FCR) of LDL apolipoprotein B (apoB) from baseline to a selected time point after rAAV administration are provided. The method involves administering to the human subject via a peripheral vein by infusion of a suspension of replication deficient recombinant adeno-associated virus (rAAV).

1 . A pharmaceutical composition suitable for peripheral vein infusion in human subjects, comprising a suspension of replication deficient recombinant adeno-associated virus (rAAV) in a formulation buffer, wherein:
 (a) the rAAV comprises a vector genome comprising AAV ITRs and a nucleic acid sequence encodes a human LDL receptor (hLDLR) operably linked to a liver specific promoter, said vector genome packaged in an AAV8 capsid;   (b) the formulation buffer comprises an aqueous solution of phosphate buffered saline and a surfactant, wherein the surfactant is at concentration of about 0.0005% to about 0.001% of the suspension;   (c) (i) the rAAV is at least about 95% free of empty capsids as determined by oqPCR or ddPCR
 (ii) the rAAV Genome Copy (GC) titer is at least 1×10 13  GC/ml; 
 (iii) the Empty:Full particle ratio is between 0:4 to 1:4; and 
 (iv) a dose of 5×10 11  GC/kg of the rAAV suspension decreases baseline cholesterol levels in a double knockout (DKO) LDLR−/−Apobec−/− mouse model of Homozygous Familial Hypercholesterolemia (HoFH) by 25% to 75%. 
   
     
     
         2 . The composition according to  claim 1 , wherein the rAAV is AAV8.TBG.hLDLR, wherein the liver specific promoter is a thyroxine binding globulin (TBG) promoter. 
     
     
         3 . The composition according to  claim 1 , wherein the formulation buffer is 180 mM NaCl, 10 mM Na phosphate, 0.001% poloxamer 188, pH 7.3. 
     
     
         4 . A pharmaceutical composition according to of  claim 1  suitable for use in treating a human subject diagnosed with Familial Hypercholesterolemia (FH). 
     
     
         5 . The composition according to  claim 1 , wherein the composition is administrable to the human subject via a peripheral vein by infusion of a suspension of replication deficient recombinant adeno-associated virus (rAAV) at a dose of (a) at least about 5×10 11  Genome Copies (GC)/kg or (b) 2.5×10 12  Genome Copy (GC)/kg to 7.5×10 12  Genome Copy (GC)/kg body weight of the human subject as determined by oqPCR or ddPCR 
     
     
         6 . The composition according to  claim 5 , wherein the rAAV is AAV8.TBG.hLDLR, wherein the liver specific promoter is a thyroxine binding globulin (TBG) promoter and the nucleic acid sequence further comprises two alpha mic/bik enhancer sequences, an intron, a polyadenylation (polyA) signal and wherein the hLDLR coding sequence has the nucleic acid sequence selected from SEQ ID NO: 2 or SEQ ID NO: 4. 
     
     
         7 . The composition according to  claim 4 , wherein the formulation buffer is 180 mM NaCl, 10 mM Na phosphate, 0.001% Poloxamer 188, pH 7.3. 
     
     
         8 . The composition according to  claim 4 , wherein the subject has been diagnosed with Homozygous FH (HoFH). 
     
     
         9 . The composition according to  claim 4 , wherein the subject has been diagnosed with Heterozygous FH (HeFH). 
     
     
         10 . A method of treating a human subject in a need thereof, wherein the method comprises administering a composition according to  claim 1  to reduce one or more of: LDL-cholesterol, total cholesterol, and/or fasting triglycerides in a human subject, and/or modifying fractional catabolic rate (FCR) of LDL apolipoprotein B (apoB) from baseline to a selected time point after an rAAV administration. 
     
     
         11 . The method according to  claim 10 , wherein the composition is administrable to the human subject via a peripheral vein by infusion of a suspension of replication deficient recombinant adeno-associated virus (rAAV) at a dose of (a) at least about 5×10 11  Genome Copies (GC)/kg or (b) 2.5×10 12  Genome Copy (GC)/kg to 7.5×10 12  Genome Copy (GC)/kg body weight of the human subject as determined by oqPCR or ddPCR. 
     
     
         12 . The pharmaceutical composition according to  claim 11 , wherein said human subject is co-treated with one or more of a PCSK9 inhibitors; a statin; niacin; ezetimibe, or bile acid sequestrants. 
     
     
         13 . A method of treating a human subject diagnosed with Familial Hypercholesterolemia (FH), comprising administering to the human subject via a peripheral vein by infusion of a pharmaceutical composition according to  claim 1 , wherein the suspension of replication deficient recombinant adeno-associated virus (rAAV) is at a dose of (a) at least about 5×10 11  Genome Copies/kg or (b) 2.5×10 12  Genome Copy (GC)/kg to 7.5×10 12  Genome Copy (GC)/kg body weight of the human subject as determined by oqPCR or ddPCR. 
     
     
         14 . The method according to  claim 13 , wherein the rAAV is AAV8.TBG.hLDLR. 
     
     
         15 . The method according to  claim 13 , wherein the formulation buffer is 180 mM NaCl, 10 mM Na phosphate, 0.001% Poloxamer 188, pH 7.3. 
     
     
         16 . The method according to  claim 13 , wherein the subject has been diagnosed with Homozygous FH (HoFH). 
     
     
         17 . The method according to  claim 13 , wherein the subject has been diagnosed with Heterozygous FH (HeFH). 
     
     
         18 . A method of reducing one or more of: LDL-cholesterol, total cholesterol, and/or fasting triglycerides in a subject, and/or modifying fractional catabolic rate (FCR) of LDL apolipoprotein B (apoB) from baseline to a selected time point after rAAV administration, said method comprising administering to the human subject via a peripheral vein by infusion of the pharmaceutical composition according to  claim 1  comprising a suspension of replication deficient recombinant adeno-associated virus (rAAV) at a dose of (a) at least about 5×10 11  Genome Copies (GC)/kg or (b) 2.5×10 12  Genome Copy (GC)/kg to 7.5×10 12  Genome Copy (GC)/kg body weight of the human subject as determined by oqPCR or ddPCR. 
     
     
         19 . The method according to  claim 18 , wherein said human subject is co-treated with one or more of a PCSK9 inhibitors; a statin; niacin; ezetimibe, or bile acid sequestrants.