| cIT was a monomer gained from chemical reaction of Epimedium flavonoid,and there was cIT in Epimedium.It had been proved that Epimedium flavonoid had activity on osteoporosis,no matter it had glycoside or not.A hypothesis was raised:cIT had activity on osteoporosis as others.We had proved that by experiments.Epimedium flavonoid selected as raw material,we prepared cIT through chemical reaction.Following,we initial investigated its activity on osteoporosis.We had found that the activity of alkaline phosphatase was enhanced by cIT.It indicated that cIT had activity on osteoporosis.However,cIT was poor water-solubility and absorption as other Epimedium flavonoid in biopharmaceutics.We increased its solubility by pharmaceutical method,and then anti-osteoporotic activity and its absorption in vivo were investigated.It also provided an according for pharmaceutical dosage forms selection.1.Epimedium flavonoid as raw material,cIT was prepared by acid hydrolysis.Structure of cIT was definites by Spectrum analysis,and it was systematically named.It suggested that cIT was an isomeride of IT,a kind of flavonoid in Epimedium,which possessed anti-osteoporotic activity.From initial recerch,we found that cIT can increase alkaline phosphatase activities in cells,and it was probably had activity on osteoporosis.2.Solubility of cIT in common solvents and buffers in different pH were determined, and its oil-water partition coefficient was determined,too.It suggested that cIT has poor solubility in both oil and water.A four-site perfused rat intestinal model was established to determin the appear permeability coefficien of cIT,and the result suggested that cIT had poor absorption in vivo(P*eff＜0.75).3.To enhance solubility of the compound,cIT microemulsion,which was clear,limpid and stabilized,was prepared.The mean diameter of microemulsion was 63.7nm,and the content of cIT was 32.02μg·mL-1.For cIT micro emulsion,it was O/W,and drug-loading rate was 2.10%4.Both entrapment rate and mean diameter as index,the single factor tests and orthogonal test were constructed to optimize the formula of cIT liposome.It was consisted of 62.5%soybean lecithin,31.25%cholesterol and 6.25%cIT.Freeze dehydration technique was used to prepare cIT freeze-dried liposome.We investigated the kind of freeze-dried protecting agent,its proportion and so on.10%mannitol was selected as its protector.5.Quality evaluation were undertaken for cIT liposome and cIT microemulsion,such as mean diameter,Zeta electric potential,morphology,and so on.The stability of preparations were also evaluated,cIT liposome was sphericity.Structure of lipid bilayer was seen with electron microscope.For cIT liposome,its mean diameter was 229.7±28.6nm,Zeta electric potential was 47.2±3.5mV,entrapment rate was 92.35±0.66%,drug-loading rate was 6.25%.To investigate stability of liposome,accelerated test was to last six months,and the result showed that mean diameter and entrapment rate keep stabilizing.6.Marrow stroma cell culture model was first used to investigate anti- osteoporotic activity for cIT and its preparations.Alkaline phosphatase activity in cell was determined by CAKP dyeing,total clone was determined by methylene blue dyeing.The results suggested that cIT liposome could significantly promote osteoblasts proliferation,enhance activity of alkaline phosphatase activity in cell,and inhibite satellite phenomenon of fat cell in osteoblasts proliferation in some degree. |
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