Preparation Of VD3 Lipsomes And Preliminary Study On Inhibition Of Photoperiodic Activity In In Vitro And In Vivo

Vitamin D3(Vitamin D3,VD3)is a fat soluble vitamin,belongs to an effect on calcium and phosphorus metabolism of hormone precursor,also known as the "sunshine vitamin." VD3 has many functions,such as the regulation of calcium and phosphorus metabolism,prevention of osteoporosis improved cardio-cerebral vascular diseases,improve immunity,and so on.But VD3 is fat soluble,insoluble in water,limiting its effect.In order to improve the bioavailability of VD3,VD3 preparations needed to design a safer and more efficient.Liposomes were composed of a layer or multi-layer gender bilayer composition and internal micro-vesicles that contain water areas,can be used as a drug delivery system,achieve the purpose of enhancing efficacy and reduced side effects.This paper VD3 highly oxidizing,prevent skin cells overproduction of free radicals,increase free radical scavenging enzyme activity,VD3 combined with Liposomes,preparation of liposomal Vitamin D3(L-VD3)to prevent the skin from the effects of aging.This article includes the following:1.VD3 in vitro establishment of analytical methodology.Establish a method for in vitro analysis of VD3-high performance liquid chromatography(HPLC),use this method to get the VD3 precision and recovery rates are in line with the requirement in vitro assay.HPLC determination of content of L-VD3 and the method of determination of L-VD3 in drug recovery in line with requirements for in vitro analysis.Established free VD3 separation by centrifugal ultrafiltration and determination of entrapment efficiency of L-VD3 method.This method can be very good separation of the free drug,easy and effective,and the determination of VD3 accuracy and recovery and are in line with the requirement in vitro assay.2.VD3 General liposome and evaluation of ethanol injection respectively,film homogenization,and evaporation method to filter the preparation method of L-VD3,compared the entrapment efficiency of liposomes and particle size to get ethanol injection method is the most suitable method for preparing L-VD3.Then respectively L-VD3 formula in the Phospholipid,lecithin and cholesterol ratio,the ratio of drugsand phosphate buffer of PH value for single factor.Filtered lecithin Phospholipid and cholesterol content,scale,lipid than response surface optimization experiment as the main subject,the optimal formula for: Phospholipid 25 mg,Phospholipid/cholesterol ratio is 4.5:1,to 2.5:l drugs.Vitamin d under conditions of optimal formulation of conventional liposomes encapsulation of theoretical value of 87.83%,the actual value is 86.79%.3.Study on the stability of VD3 liposomes.Based on the temperature,light,freeze-thaw conditions are factors that influence the stability of L-VD3 test.Which temperature in 4-40 ? of conditions Xia,package seal rate basic not occurred too big changes,stability better,suitable save L-VD3;light on L-VD3 of stability no too much effect,so can description VD3 in preparation into fat quality body zhihou light of stability greatly improve has,can without avoidance light save;frozen thaw conditions on L-VD3 of stability also no too much effect,can description VD3 made fat quality body Hou on temperature of mutation stability better,also can is good of adapted temperature of changes.In addition,VD3 in vitro release test is conducted,found L-VD3 in the units trapped in the skin dose to182.13 ? g/cm2 and VD3 units trapped in the skin dose is 113.82 ? g/cm2 interception doses much larger than the L-VD3 units VD3 and L-VD3 transdermal delivery of less than VD3 and VD3 experience more beneficial to skin absorption of lipids.Then they accelerated and long-term experiments,found L-VD3 of entrapment and their appearance has not changed much,explain the stability of L-VD3.4.L-VD3 study on the efficacy of skin photoaging model is established.After sunburn treatment a marked folds of skin,redness,spots,varying degrees of ageing phenomena such as white blocks,model successful.Drug treatment has a time zhihou observation big rat of skin State,found smear L-VD3 and positive control drug Hou of big rat skin basically recovery normal,no obviously of Blaze folds damaged,phenomenon,smooth has elastic;smear has VD3 solution of big rat skin status also has must of improved,although still has some macular red points,but not many;and smear PBS solution of negative control group big rat skin still has Erythema bleeding blaze folds,light aging phenomenon,no get improved.Zhihou will the Group big rat of skin for HE dyeing,observation slice of electric mirror pictures,found smear L-VD3 and positive control drug of big rat epidermal leather structure basic full,internal fibrous tissue,and glands,arrangement uniform,and normal of big rat skin organization slice State almost;smear has VD3 solution of big rat epidermal leather structure slightly has change,internal fibrous tissue,and glands individual deformation,but overall Shang compared ordered arranged;and smear PBS solution of negative control group big rat epidermal leather serious layered,Blurring the boundaries between,fibers are also disorganized and even deformation of internal glands,skin injury situation did not improve.These results suggest,L-VD3 can be effective in the prevention of skin ageing phenomena,have a more significant effect.