Preparation Of Danshensu-Phospholipid Complex And Its Evaluation

Danshen to labiatae plants of the genus Salvia danshen (Salvia miltiorrhiza Bge) dry root and rhizome, which has effect such as dispelling pains and stasis blood menstruation and blessing bother, used to menstruation to be not moved, the sting of a chest, sleepless, conditions such as angina. Danshensu, isolated from active ingredients of water soluble part, is a major ingredient of salvia miltiorrhiza extract. Pharmacological experiment proves that danshensu has function such as expansion of heart blood-vessel, gathered resist platelet adhesion to the removal of oxygen free radical. According to Biopharmaceutics Classification System (BCS), as a class Ⅲ compound, the oral absorption was limited because of the bad lipophilicity of danshensu. The phospholipid complex is one of preparation technologies to improve drug bioavailability in recent years, this study intends to use danshensu as model drug, with the combination of phospholipid and danshensu as the evaluation index, using the solvent method to prepare danshensu phospholipid complex, in order to improve its bioavailability and enhance the affinity of danshensu for cellular membrane. On the basis of the preparation of danshensu phospholipid complex, prepared its soybean oil, palm oil, and olive oil solution, to further enhance its lipophilicity and improve the bioavailability and absorption in vivo.In this paper, the preparation technology of phospholipid complex, the physic-chemical property and its pharmacokinetic were studied systematically and deeply, which could provide reference for the research and develop preparations of danshensu. The paper includes the following several parts:1. Dansehnsu phospholipid complex preparation, characterization, and the physic-chemical properties(1) Optimize of the preparation technology of danshensu phospholipid complexA reasonable criterion of accurate evaluation-complexing rate established during preparation of danshensu phospholipid complex, and the reaction solvent, concentration of reactants, weight ratio of phospholipids and danshensu (w/w), reaction time and reaction temperature were examined. On the basis of the single factor investigation, we select the concentration of reactants, weight ratio of phospholipids and danshensu (w/w), and reaction temperature, which influence notable of complexing rate, and then a central composite design approach was employed for process optimization.The optimization process of the final verification, have a good correlation between predicted values and measured values. The best preparation technology for:reaction of anhydrous ethanol solvent; Concentration of reactants to 45 mg/100 mL; Phospholipids-drug weight ratio (w/w) for 2:1, response time is 2 h, the reaction temperature for 57℃.(2) Characterization of danshensuphospholipid complex and the physic-chemical propertiesBy differential scanning calorimetry analysis (DSC), X-ray diffraction analysis (XRD) map, we can see that in phospholipid complex with endothermic peak, loss of crystal characteristics, and shows that the phospholipid complex exists in the form of amorphous, preliminary confirmed the formation of phospholipid complex. By infrared (IR) spectrum can see that, compared with danshendu and the mixture, there was no new chemical bonds to form in phospholipid complex, speculation in complex was combined danshensu with phospholipids by hydrogen bonding or van der Waals force. Oil/water partition coefficient (P) studies show that, compared with danshendu and the mixture, phospholipid complex was in the oil-water partition coefficient in different pH of PBS solution are increased.2. The research on pharmacokinetic of danshensu phospholipid complex in rats(1) To establish accurate, reliable and easy method for determination of the danshensu content in plasma of ratsChromatographic conditions:Waters ACQUITY UPLC BCH C18 column (1.7μm and 2.1 x 100 mm); Protect column:BCH C18 column (1.7μm and 2.1 x 5 mm); Mobile phase:(A) 0.1% formic acid water and acetonitrile (B) and gradient elution,0-0.5 min 90% A,10% B,0.5-1.5 effect is smaller, yet to be further improved.3. The pharmacokinetic study in rats of gastric gavage the oil solution of danshensu phospholipid complexThe 24 healthy male SD rats were randomly divided into four groups, respectively by gastric gavage with saline solution and the oybean oil, coconut oil, palm oil solution of danshensu phospolipid complex. The blood concentration at different intervals was tested by LC-MS/MS, and DAS software is applied to estimate pharmacokinetic parameters. Results show that compared with saline solution of danshensu phospholipid compound, the bioavailability of soybean oil/palm oil solution were improved significantly. But the bio availability of the olive oil solution is less than that with phospholipid complex, the reason needs further research.