Composition for increasing expression of blood coagulation factor gene, comprising core-shell structured microparticles as active ingredient

Abstract

The present disclosure relates to a composition for increasing the expression of a blood coagulation factor gene, which contains core-shell structured microparticles as an active ingredient. When administered in vivo along with blood coagulation factor VIII gene or a variant gene thereof, the composition for increasing the expression of a blood coagulation factor gene of the present disclosure can increase the expression of the gene by at least 30%. When administered along with a gene therapeutic agent, the composition can achieve a therapeutic effect even with a very small amount of a gene, and thus is useful.

1 . A method for increasing the expression of a blood coagulation factor gene in a subject in need thereof comprising administering to the subject the blood coagulation factor gene and a composition comprising core-shell structured microparticles, wherein
 the core is sulfur hexafluoride or perfluorobutane as a biocompatible gas, and the shell is a lipid or a derivative thereof, and   the blood coagulation factor gene is one or more genes selected from a human blood coagulation factor VIII gene or a variant gene thereof.   
     
     
         2 . The method for increasing the expression of a blood coagulation factor gene according to  claim 1 , wherein the lipid is one or more selected from a group consisting of a simple lipid, a phospholipid, a glyceroglycolipid, a sphingoglycolipid, a cholesterol and a cationic lipid. 
     
     
         3 . The method for increasing the expression of a blood coagulation factor gene according to  claim 2 , wherein the phospholipid is selected from a group consisting of a phosphatidylcholine derivative, a phosphatidylethanolamine derivative, a phosphatidylserine derivative, a diacetylated phospholipid, L-α-dioleyl phosphatidylethanolamine, diolein, phosphatidic acid, phosphatidylglycerol, phosphatidylinositol, lysophosphatidylcholine, sphingomyelin, a polyethylene glycolated phospholipid, egg yolk lecithin, soy lecithin and a hydrogenated phospholipid. 
     
     
         4 . The method for increasing the expression of a blood coagulation factor gene according to  claim 2 , wherein the glyceroglycolipid is selected from a group consisting of sulfoxyribosyl glyceride, diglycosyl diglyceride, digalactosyl diglyceride, galactosyl diglyceride and glycosyl diglyceride. 
     
     
         5 . The method for increasing the expression of a blood coagulation factor gene according to  claim 2 , wherein the sphingoglycolipid is galactosyl cerebroside, lactosyl cerebroside or ganglioside. 
     
     
         6 . The method for increasing the expression of a blood coagulation factor gene according to  claim 2 , wherein the cationic lipid is selected from a group consisting of 1,2-dioleoyl-3-trmethylammonium propane (DOTAP), N-(2,3-dioleyloxypropan-1-yl)-N,N,N-trmethylammonium chloride (DOTMA), 2,3-dioleyloxy-N-[2-(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminium trifluoroacetate (DOSPA), 1,2-dimyristyloxypropyl-3-dimethylhydroxyethylammonium bromide (DMRIE), 1,2-dioleoyloxypropyl-3-diethylhydroxyethylammonium bromide (DORIE) and 3β-[N—(N′N′-dimethylaminoethylhy)carbamoyl]cholesterol (DC-Chol). 
     
     
         7 . The method for increasing the expression of a blood coagulation factor gene according to  claim 1 , wherein the blood coagulation factor VIII is composed of an amino acid sequence represented by SEQ ID NO 1, and the variant of blood coagulation factor VIII is composed of an amino acid sequence represented by SEQ ID NO 3. 
     
     
         8 . The method for increasing the expression of a blood coagulation factor gene according to  claim 1 , wherein the composition increases the expression of the blood coagulation factor gene by 30% or more when the composition is administered along with the human blood coagulation factor VIII gene or a variant gene thereof. 
     
     
         9 . A method for preventing or treating a bleeding disorder or bleeding in a subject, comprising administering to the subject the blood coagulation factor gene and a composition comprising core-shell structured microparticles, wherein
 the core is sulfur hexafluoride or perfluorobutane as a biocompatible gas, and the shell is a lipid or a derivative thereof, and   the blood coagulation factor gene is one or more genes selected from a human blood coagulation factor VIII gene or a variant gene thereof.   
     
     
         10 . The method for preventing or treating a bleeding disorder or bleeding according to  claim 9 , wherein the bleeding disorder is hemophilia A, hemophilia induced or complicated by an inhibitory antibody against blood coagulation factor VIII or blood coagulation factor VIIIa, or hemophilia B. 
     
     
         11 . The method for preventing or treating a bleeding disorder or bleeding according to  claim 9 , wherein the bleeding disorder or bleeding is one of neonatal coagulopathy, severe liver disease, thrombocytopenia, congenital deficiency of factor V, VII, X or XI, and von Willebrand disease having an inhibitor against von Willebrand factor. 
     
     
         12 . The method for preventing or treating a bleeding disorder or bleeding according to  claim 9 , wherein the bleeding is caused by blood loss associated with high-risk surgical procedures, traumatic blood loss, blood loss caused by bone marrow transplantation or blood loss caused by cerebral hemorrhage.