Studies On The Vinorelbine Intravenous Emulsion

Vinorelbine can interact tumour cell metabolism by effecting microtubule protein of tumour cell,and it is used to cure the Lymphocytic leukemia,Hodgkin's lymphoma and non-Hodgkin's lymphoma,Bronchogenic carcinoma,Soft tissue sarcoma,Neuroblastoma and so on in clinical.Intravenous injection emulsion has slow and controlled release,targeting function,and there is advantage of reducing the side effect of medicine at the same time.In domain of the antitumor,Antimicrobial,resisting cerebrovascular disease, intravenous emulsion has unique curative effect.Vinorelbine which is short of water-solubility,large of irritation is prepared into intravenous emulsion. This research can raise the alternative of vinorelbine to different tissue, raise the concentration in the lung and the lymph,thus it offers new clinical applied route of vinorelbine.The study can also provide theoretical references for other studies of TCM targeting drug delivery system and promote the development of TCM new form drug delivery system.The paper consists of four parts.1.Technology research of VRB intravenous emulsion1.1 Basic research before prescription1.1.1 The Determination of the apparent octanol/water distribution coefficient of vinorelbineChromatogram condition refering the relevant documents was established as follows:column was Gemini C₁₈(250mm×4.6mm,5μm,Phenomenex),mobile phase was acetonitrile:0.06mol/L potassium dihydrogen phosphate(35:65),column temperature 30℃,sample size5μl,wavelength 215nm,flow rate 0.8ml/min and number of theoretical plates can't be lower than 3000 according to peak area of VRB.It shows that the apparent octanol/water distribution coefficient of vinorelbine is different under the condition of different pH circumstances. The apparent octanol/water distribution coefficient of vinorelbine is high when pH＞7.0.It indicats that VRB is apt to store as the molecule state in alkaline solution.In conclusion,we should guarantee the preparation is under the condition of pH＞7.0 to obtain the high rate of envelopment.1.1.2 Determination of the solubility of VRBThe solubility of VRB was determined in different solvent according to the assay of solubility regulated in pharmacopoeia of Chinese 2005 edition. The solubility of VRB is relatively high in ether and chloroform,and low in light petroleum,indicating that VRB is liposolubility.The solubility of VRB in oil is 60.07mg/ml,which helps the medicine dissolving among the oil,and is favorable to the preparation of intravenous emulsion.1.2 Optimization of prescriptionOn the foundation of investigating in the single factor,central composite design/response surface methodology was introduced to optimize the dosage of the oil,phospholipid and the poloxamer regarding the stability parameter K_E and mean diameter of emulsion.The results were determined that the amount of oil was 9.0%,phospholipid 2.25%and poloxamer 1.51%.Theoretical value of average particle size and the stability parameter K_E was 126.48nm and 19.92% whereas experimental value was 141.3nm and 18.80%respectively.1.3 Preparing craft of VRB intravenous emulsionIn order to gain the best method of making the original emulsion,the influences of different factors which were the times of ultrasonic,the intensity of ultrasonic,ultrasonic time and the mixing temperature of water and oil were evaluated by orthogonal-designing method using L₉(3⁴)table.In the studies,the best preparing craft was made:VRB was dissolved in the soya bean oil;phospholipid,poloxamer,glycerine were dissolved in the water,the water and the oil were heated to 60℃on the water-bath.The oil was added into the water slowly,and the original emulsion was made under the conditon of 400kHz intensity,ultrasound 30min.2 Quality standard study of VRB intravenous emulsion2.1 The assay method of VRB in the intravenous emulsionWe set up the assay method of VRB in the intravenous emulsion withHPLC, and chromatogram condition is the same as the contents of 1.1.The best extracting technology of VRB in emulsion was established as follows:taking emulsion 2g definitely,with 0.1mol/L NaOH2.0ml,adding ether 30ml, ultrasounding 30 minutes,centrifuging 10 minutes,the ether was stored in the certain device,the remainder was added 0.1mol/L NaOH2.0ml,ether 30ml, ultrasounded 30 minutes,centrifuging 10 minutes,combining ether liquids, and evaporating ether to dryness on the aqueous bath,metering volume to 2ml with mobile phase definitely.2.2 The physical and chemical property of VRB emulsionThe distribution in the internal body and pharmacokinetics course were changed after the medicine is prepared into intravenous emulsion.This change is related to the fact that the composition of the membrane in emulsion,particle size,electricity on the surface of membrane,viscosity of the preparation,and so on.Physical and chemical property of surface on emulsion had been done,and the result was that particle size was 121.9nm,the size of Zeta was 35.58my,and the viscosity was 1.83mPa·s.2.3 The research of stability of VRB emulsionThere is no change in the stability parameter K_E and average particle size of emulsion after shaking experiment,centrifugalizating experiment,freezing-heating experiment,accelerating experiment,thermocompression sterilizing experiment.The emulsion holded one storey constantly in various stability experiments,indicating that the stability of emulsion is good.3 Security research of VRB emulsionCompared with VRB injection,the safety of intravenous emulsion was investigated:LD₅₀of VRB injection and intravenous emulsion were 1.53mg/kg and 4.19 mg/kg respectively.Haemolysis test were conducted.The result of subacute toxicity indicated that the side effects of emulsion were diminished. The research of haemolysis test indicated that intravenous emulsion of VRB had no harmful effect on the blood,while VRB injection has slight haemolysis functiom.All of this is caused by that VRB is dissolved in the oil in emulsion, and degreasing the chance of direct contact to the erythrocyte.4 Study on In-vivo Pharmacokinetics and disposition in tissues of VRB emulsion4.1 Study on In-vivo Pharmacokinetics of VRB emulsionPharmacokinetics of VRB in rabbit were studied in this paper.The high performance liquid chromatographic(HPLC)method using a reverse-phase column and UV detector was adopted for the determination of VRB and coculine in plasma to study the pharmacokinetics of VRB in rabbit.The concentration-time curve of rabbits after iv VRB can be fitted to two-compartment model with software 3P87.It indicated that VRB emulsion is longer biologic half-life comparing with VRB injection.4.2 Study on distibution in tissues of VRB emulsionorganization distributing experiment indicated:The concentration in the lung is changed obviously afterâ…³VRB emulsion.The quantitative evaluation parameter of TDDS in the lung was 3.0043,however it was less than 1.000 in the other organs.It can indicate that the emulsion increase the concentration of VRB in the lung,and decrease the drug level in the other organs,so it cut down the side effect.The quantitative evaluation parameter was 0.4106 and 0.2933 in the liver and kidney,which caused by that VRB was metabolized in the liver and kidney mainly,and the preparation of VRB emulsion changed the distibution of the medcine in tissues so as to degrade the concentration in them.