Preparation And Preliminary Efficacy Test Of Lovastatin Mediated By GM1 Modified Recombinant High-density Lipoprotein

Poor lifestyle,few physical exercise,and high-fat/high-sugar/high-protein intake generally results in “three high” symptoms and even cardiovascular diseases(e.g.atherosclerosis).The spread of healthy lifestyle and the research/development of related drugs have been getting more and more attention.Lovastatin(LT)is one of the most widely used statins for the treatment of cardiovascular diseases.However,it has many disadvantages,e.g.poor solubility in blood,short duration of presence in the systemic circulation,no site-targeting within the body,no sustained release,among others.Therefore,a variety of strategies have been developed to solve these shortcomings,among which the lipoprotein-based delivery system for statins is potentially promising.Lipoproteins,a class of endogenous lipid-protein complexes in human blood,have a phospholipid monolayer which forms a hydrophobic core and a hydrophilic shell and the inlaid apolipoproteins which can recognize/target specific tissues or cells for cholesterol delivery.Among them,high-density lipoprotein(HDL),containing apolipoprotein A-1(ApoA-1)which can be recognized by scavenger receptors,transports cholesterol from peripheral tissues to the liver for metabolism and elimination.Moreover,HDL has lipid-lowering and anti-atherosclerotic effects and is commonly known as "vascular scavenger." Therefore,recombinant HDL(rHDL),in combination with other modifications,can achieve efficient targeted delivery,sustained release,greater bioavailability,and other purposes.The use of rHDL as a drug carrier to transport statins is a hotspot in recent years,but its retention time in the systemic circulation,sustained release,and other properties still needs to be improved.Utilization of monosialoglycans(e.g.GM1)to modify r HDL is expected to achieve this goal.Ganglioside GM1 is one of the endogenous cell membrane components.It has been reported that liposomal surface modification of GM1 can prolong its residence time in blood and reduce liposome uptake by liver.In this study,for the first time,GM1-modified rHDL was used as a drug carrier to deliver lovastatin,and the drug-carrier complex(GM1-LT-rHDL)was prepared and identified,as well as preliminary test of its anti-atherosclerotic efficacy.(1)The lipid carrier was prepared first by thin film dispersion method,further modified with GM1 and ApoA-1,and then loaded with the drug to obtain GM1-LT-rHDL.Exogenously added ApoA-1 can target the drug to the damaged plaque area whereas exogenously added GM1 helps to improve sustained release of the drug,prolong its retention time in blood,and reduce its liver uptake.(2)The morphology and size of GM1-LT-rHDL were detected by Zeta particle size analyzer and transmission electron microscope.The particle size was 117.1 ± 3.6 nm and the zeta potential was 41.97 ± 1.57 mV.(3)Using silicon beads and macrophages as models,by applying laser confocal microscopy and flow cytometry,it was proved that GM1-LT-rHDL contains GM1 and ApoA-1 molecules,confirming the successful construction of GM1-LT-rHDL.(4)The encapsulation efficiency and loading content of the drug are 76.55 ± 0.41% and was 3.83 ± 0.02%,respectively detected by high performance liquid chromatography.(5)The in vitro drug release experiments confirmed the sustained drug release of GM1-LT-rHDL.(6)The in vitro cellular experiments confirmed that GM1-LT-rHDL can be recognized by the scavenger receptor SR-BI on the surface of macrophages and that GM1-LT-rHDL has potential anti-atherosclerotic effect by inhibiting the Ox-LDL-induced transformation of macrophage into foam cells.(7)By using atherosclerotic model mice(ApoE-/-mice fed with high fat),it was certified that GM1-LT-rHDL has blood lipid-lowering and anti-atherosclerotic effects by detecting blood lipids with automatic biochemical analyzer and imaging the atherosclerotic plaques of the aorta stained with oil red O.In this study,the preparation and identification of the drug-carrier complex(GM1-LT-rHDL)were successfully carried out,and the efficacy of anti-atherosclerosis was preliminarily verified.However,the metabolism of GM1-LT-rHDL in the animal and its distribution in various tissues need further studied in the future.The drug carrier system based on GM1-modified rHDL can not only transport lovastatin and other statins for the treatment of cardiovascular diseases(e.g.arteriosclerosis)but also work as a carrier system for other water-insoluble drugs for the treatment of other diseases which can be further investigated in the future.