| Huoxuezhitong pach has an application history for a long time and its therapy effect is definite,but there are many disadvantages in the current drug delivery system(DDS)of it to limit its clinical application.The application of novel polymeric materials can improve the bioavailability of drugs and be lower irritability,which has broad prospects for development.Hence,this group early prepared Huoxuezhitong gel patch and microemulsion gel,but the arriving at subcutaneous of drugs in transdermal preparation is the precondition of its therapeutic effect,so it is necessary to study the transdermal permeability and local pharmacokinetics of the three kinds of transdermal drug delivery.At present,Franz diffusion cell method is easy to operate and its test condition is simple to control,widely used in the study of transdermal permeation;and due to its advantages of in vivo,real time,online sampling for a long time and so on,microdialysis(MD)plays an important role in the study on pharmacokinetics of transdermal drug delivery.Hence,this study was designed to compare the percutaneous permeability and in vivo pharmacokinetics of Huoxuezhitong rubber patch,gel patch and microemulsion gel with the index constituent of Pae,Eug and Pip,using Franz diffusion cell method and MD,and to provide new idea for the development of safe,high-efficient and novel DDS.1.Study on in vitro transdermal permeability of Huoxuezhitong preparations(1)The establishment of determination method for drugs in receptor liquidA determination method of concentration of Pae,Eug and Pip in receptor liquid was established by HPLC,qualified by evaluating its specificity,linearity,precision and stability of samples.(2)The selection of receptor liquidThe receptor liquid was optimized by taking Q24,Jss and skin irritation as index.And the results showed that the Q24 and Jss were higher using PEG400-Et-NS(3:3:4)as receptor liquid than PG-Et-NS(3:3:4)and Et-NS(3:7),and skin irritation of PEG400-Et-NS(3:3:4)was smaller than PG-Et-NS(3:3:4).Hence,PEG400-Et-NS(3:3:4)was preferred as receptor fluid for transdermal permeability study.(3)The comparation study on percutaneous permeability of three kinds of Huoxuezhitong preparationsIn vitro Franz diffusion experiment was applied to assess the percutaneous penetration characteristic and regularity of Huoxuezhitong rubber patch,gel patch and microemulsion gel with the index of Qn and Jss.And the results showed that the Q24 of Pae,Eug and Pip in Huoxuezhitong rubber patch were respectively(2.40±0.20),(19.9±2.46)and(0.77±0.24)?g·cm-2 and the Jss were respectively 0.18,1.26 and 0.02?g·cm-2·h-1;the Q24 of Pae,Eug and Pip in Huoxuezhitong gel patch were respectively(8.77±0.42),(96.0±5.5)and(5.53±0.11)?ig·cm-2 and the Jss were respectively 0.63,6.12 and 0.23 ?g·cm-2·h-1;the Q24 of Pae and Eug in Huoxuezhitong microemulsion gel were respectively(2.59±0.V9)and(28.5±2.3)?g·cm-2 and the Jss were respectively 0.21 and 1.94?g·cm-2·h-1.By the results,for Pae in gel patch,the Q24 and Jss value was superior to rubber patch and microemulsion gel;for Eug in patches,the the sequence was gel patch,microemulsion gel,and rubber patch based on value of Q24 and Jss from large to small;for Pip,the Q24 and Jss value in gel patch was higher obviously than rubber patch.As was above-mentioned,gel patch could significantly improve the transdermal permeation rate and amount of Pae,Eug and Pip,while permeation enhancing effect of microemulsion gel was not obvious.The percutaneous permeation mechanism of three kinds of preparations was studied by fitting percutaneous penetration curve respectively with Zero order model,First order model,Higuchi model,Hixson-Crowell model and Korsmeyer-Pappas model,and it turned out that:? the correlation coefficients of Zero model fitting equation of Pae,Eug and Pip in the three preparations were higher than those of First order model,indicating that Pae,Eug and Pip in the three preparations through the skin were a diffusion process and it was irrelevant to the content of them in patch;? the correlation coefficients of Higuchi model fitting equation of Pae,Eug and Pip in Huoxuezhitong rubber patch were respectively 0.9632,0.9909 and 0.8963;the correlation coefficients of Hixson-Crowell model fitting equation of Pae,Eug and Pip in Huoxuezhitong rubber patch were respectively 0.7578,0.8115 and 0.4715;the correlation coefficients of Korsmeyer-Pappas model fitting equation of Pae,Eug and Pip in Huoxuezhitong rubber patch were respectively 0.9745,0.9860 and 0.8431,which indicated that percutaneous penetration mechanism of three components in rubber patch belonged to the diffusion process.? The correlation coefficients of Higuchi model fitting equation of Pae,Eug and Pip in Huoxuezhitong gel patch were respectively 0.9779,0.9892 and 0.9643;the correlation coefficients of Hixson-Crowell model fitting equation of Pae,Eug and Pip in Huoxuezhitong gel patch were respectively 0.7830,0.8140 and 0.9400;the correlation coefficients of Korsmeyer-Pappas model fitting equation of Pae,Eug and Pip in Huoxuezhitong gel patch were respectively 0.9802,0.9850 and 0.9967,and the slope of this equation were 0.6616,0.7889 and 1.1506,which indicated that percutaneous penetration mechanism of Pae and Eug in gel patch belonged to the corrosion and diffusion process,while Pip belonged to corrosion effect.? The correlation coefficients of Higuchi model fitting equation of Pae and Eug in Huoxuezhitong microemulsion gel were respectively 0 0.9619 and 0.9823;the correlation coefficients of Hixson-Crowell model fitting equation of Pae and Eug in Huoxuezhitong microemulsion gel were respectively 0.7398?0.7783;the correlation coefficients of Korsmeyer-Pappas model fitting equation of Pae and Eug in Huoxuezhitong gel patch were respectively 0.962 and 0.9713,and the slope of this equation were 0.759 and 0.865,which indicated that percutaneous penetration mechanism of Pae and Eug in microemulsion gel patch belonged to the diffusion and corrosion effect,and the later was major.2.The establishment of the percutaneous microdialysis method(1)The study on probe adsorptionDue to the strong lipid solubility of the active ingredients of the Huoxue Zhitong preparations,there is a problem of probe adsorption.So,in this part,the perfusion fluid was selected referring the probe absorbability as index,and the results showed that there was little Pae,Eug and Pip exist in probe with the employment of normal saline containing 30%alcohol.(2)The study on factors affecting the recovery of probeIn this part,in vitro and in vivo probe stability and repeatability of the recovery of the three components were studied,and their influence factors of concentration,calibration approaches were also investigated.The results indicated that the in vivo and vitro probe recovery coefficient of three components was stable up to 8 h and constant after changing the solution containing drug to a blank perfused for three times.And the in vitro delivery of microdialysis probe of Pae,Eug and Pip with a range of concentration were respectively(45.07±4.66)%,(27.82±2.95)%and(41.3±3.96)%,which performed concentration independent.The similar delivery was calibrated by dialysis,retrodialysis and no-net flux.Hence,the concentration of analyses in aim site could be calibrated by in vitro or in vivo recovery experiment.And it was realizable for microdialysis method built by this study to be applied to the research on percutaneous pharmacokinetics of Huoxuezhitong preparations.3.Study on transdermal pharmacokinetics of Huoxuezhitong preparations(1)The establishment of determination method for drugs in receptor liquidA determination method of concentration of Pae,Eug and Pip in dialysate was established by UPLC-MS,qualified by evaluating its specificity,linearity,precision,and matrix effect.(2)The comparation study on transdermal pharmacokinetics of three kinds of Huoxuezhitong preparationsIn this part,transdermal microdialysis technique was used for comparative study on transdermal pharmacokinetics of Huoxuezhitong rubber patch,gel patch and microemulsion gel,with Cmax?Tmax?AUC?t1/2 and as indicators.And it was turned out that:? the Cmax of Pae,Eug and Pip in Huoxuezhitong rubber patch were respectively(12.59±3.29),(18.87±7.11)and(1.80±1.22)ng·mL-1,the Tmax were respectively(186.7±48.4),(240.0±138.6)and(488.0±150.7),the AUC 0-760 were respectively(4.85±1.83),(6.92±3.57)and(0.80±0.46)?g·mL-1·min-1,the t1/2 were respectively(201.1±95.3),(218.2±123.3)and(369.6±143.3)min,and the MRT were respectively(434.7±97.8),(444.2±167.9)and(842.6±142.0)min;?the Cmax of Pae,Eug and Pip in Huoxuezhitong gel patch were respectively(83.46±27.09),(1269.4±884.7)and(0.50±0.20)ng·mL-1,the Tmax were respectively(166.7±39.3),(120.0±0.0)and(184.0±45.6),the AUC 0-760 were respectively(34.79±11.15),(375.7±208:4)and(0.20±0.12)?g·mL-1·min-1,the t1/2 were respectively(372.9±87.1),(356.2±182.6)and(391.4±150.2)min,and the MRT were respectively(625.3±136.9),(580.9±232.3)and(658.4±230.2)min;?the Cmax of Pae and Eug in Huoxuezhitong microemulsion gel were respectively(33.88±11.40)and(166.4±60.3)ng·mL-1,the were respectively(133.3±20.7)and(88,0±17.9)?the AUC 0-760 were respectively(11.09±4.46)and(35.29±17.98)?g·mL-1·min-1,the t1/2 were respectively(275.2±91.6)and(227.3±177.1)min,and the MRT were respectively(461.9±127.0)and(379.8±254.8)min.By the results,? For Pae,Cmax in gel patch was 2.5 times of microemulsion gel and 6.6 times of rubber patch,AUC 0-760 in gel patch was 3.1 times of microemulsion gel and 7.2 times of rubber patch,t1/2 in gel patch was 1.4 times of microemulsion gel and 1.9 times of rubber patch,MRT in gel patch was 1.4 times of microemulsion gel and rubber patch;Tmax in rubber patch was 1.1 times of gel patch and 1.4 times of microemulsion gel.? For Eug,Cmax in gel patch was 7.6 times of microemulsion gel and 67.3 times of rubber patch,AUC 0-760 in gel patch was 10.6 times of microemulsion gel and 54.3 times of rubber patch,t1/2 in gel patch was 1.6 times of microemulsion gel and rubber patch,MRT in gel patch was 1.5 times of microemulsion gel and 1.3times of rubber patch;Tmax in rubber patch was 2.0 times of gel patch and 2.7 times of microemulsion gel.? For Pip,Cmax in rubber patch was 3.6 times of gel patch,Tmax in rubber patch was 2.7 times of gel patch,AUC 0-760 in rubber patch was 4.1 times of gel patch,MRT in rubber patch was 1.1 times of gel patch,and t1/2 in gel patch was 1.4 times of gel patch.It revealed that the rubber patch has controlled-release effect on Pip and Eug,and gel patch could promote significantly the percutaneous absorption of Pae and Eug.In addition,for Pae,gel patch could extend MRT,but for Pip,the traditional rubber patch has obvious advantage in the aspect of enhancing penetration. |
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