| Objective:Docetaxel is a common used anti-cancer drug, but it isn’t convenient for clinical because of its highly hydrophobic. In order to improve its solubility in water, we synthesized a new diblock polymer composed of mPEG and PLA as a drug delivery carrier. mPEG segment is hydrophilic, and PLA segment is hydrophobic, so the amphiphilic polymers can self-assemble into micelles in aqueous solution, and the hydrophobic core serves as reservoir for docetaxel.Methods:mPEG-b-PDLLA diblock polymer was synthesized by a ring opening polymerization of D/L-lactide and mPEG2000under the catalysis of stannous octoate. Structure of the synthesized polymers was confirmed by1H-NMR and MALDI-TOF-TOF-MS. The value of CMC was determined by fluorometric method.The film-hydration method was used for preparation of drug loaded micelles. Synthesis of drug loaded micelles were optimized by single factor test, and studied the process reproducibility of best prescription. The morphology of drug loaded micelles prepared by optimum formulation was observed by transmission electron microscopy (TEM). The zeta potential and size distribution of the particles were also assayed. The drug loading encapsulation were determined by High Performance Liquid Chromatography (HPLC). The release of Duopafei and drug loaded micelles in PBS buffer (pH=7.4) were investigated. The hemocytolysis of Duopafei and drug loaded micelles were observed by means of the hemolytic test method in vitro.In vitro evaluation of DTX-PM:The anti-proliferation effect of blank micelles, Duopafei and DTX-PM were investigated by wound healing assay. MTT assay was done to evaluate the cytotoxicity of blank micelles, Duopafei and DTX-PM. Then, the ratios of apoptotic cells induced by blank micelles, Duopafei and DTX-PM were measured using the flow cytometry. By the way, cellular uptake of mPEG-b-PDLLA/C6was observed by laser confocal.Result:’H-NMR showed the average molecular weight was about3300, and the CMC was4.677μg/mL. The average particle diameter was (16.2±0.28) nm, PDI value was0.107±0.026, the average zeta potential was-1.35mv. By HPLC, the encapsulation efficiency of docetaxel was (89.6%±0.22)%, and the drug loading capacity was (14.01%±0.16)%. The release experiment shows that the drug release stably and adequately from micelles. Red blood cells of guinea pig appeared hemolytic resistivity to DTX-PM.In vitro Evaluation of DTX-PM:The moving ability and invasion of the4T1cells were significantly inhibited by DTX-PM. MTT result showed that low concentrations of the drug-loaded micelles have higher cell proliferation inhibition rate than the commercially available docetaxel, docetaxel-loaded mPEG-b-PDLLA nanoparticles have good inhibition activity in vitro.Conclusions:As a whole, this study designed a series of the PM loaded with DTX, and DTX-PM showed much better therapeutic effect against4T1cells. Moreover, this study revealed the great potential of the PM for the treatment of malignant breast cancer, which may provide new strategy in the development of nanomedicine for diagnosis and therapy for cancer. Objective:Active ingredients of this product, namely TZT-9, is extracted from Tripterygium wilfordii. It provides fundamental basis for developing dosage form which is safe, effective, stable and controlled. This study consists of molding craft and quality standard of dosage form.Methods:We systematically studied the molding craft and quality standard of dosage form. In order to improve the process adaptability, we adopt the method of powder tableting directly. Then, the formulation of TZT-9tablets were optimized by determining the main content of medicine, absorbent, filler, disintegrating agent and other factors. Moistureproof film coating was utilized to prevent TZT-9tablets appear the severe problem of moisture absorption. In order to investigate the technological reproducibility of TZT-9tablets, so we prepared three batches of pilot scale TZT-9tablets, inspected the quality standard, and determined the content of TZT-9in TZT-9tablets. Filmed coated TZT-9tablets were further packed with blister for long-term preservation.The study of quality standard research was in accordance with "Chinese Pharmacopoeia"2010edition of ID in Appendix2tablets under the relevant provisions while establishing the method of content determination of TZT-9tablets. At the same time, we established the quality standard method for content determination of TL-1, TL-2and TL-3. Respectively, the determination of the TL-1, TL-2and TL-3intra-day precision assay, inter-day precision assay, and solution stability, recovery rate, the minimum quantitative limit and the determination of the minimum detection limit were all studied.Results:According to test results of final TZT-9tablets of the optimal prescription for every1000pieces were that TZT-9extract0.25g, Microcrystalline cellulose37.66g, direct pressure accessories75.34g, carboxymethyl starch sodium4.8g, silicon dioxide0.75g, magnesium stearate1.2g. The tablets were all in smooth shape, good formability. Coat for three batch of pilot enlarge tablets, every piece of coating were weight gain of3%.Analytical linearity of TL-1was obtained for the method over the concentration range of0.196-2.94μg and the correlation coefficient r2=0.9998. The relative standard deviation (RSD) was1.42%for intra-day precision assay (n=6). The RSD of high concentration, median concentration and low concentration were8.60%,1.75%,4.24%for inter-day precision assay (n=5) respectively. The RSD of high concentration, median concentration and low concentration were102.39%,100.34%,100.45%for relative recovery(n=3). The limit of detection (LOD)(S/N=3) was about3.27ng, and the limits of quantification was1.31ng.Analytical linearity of TL-2was obtained for the method over the concentration range of0.194-3.88μg and the correlation coefficient r2=0.9996. The relative standard deviation (RSD) was0.38%for intra-day precision assay (n=6). The RSD of high concentration, median concentration and low concentration were1.48%,6.44%,2.17%for inter-day precision assay (n=5) respectively. The RSD of high concentration, median concentration and low concentration were100.15%,98.66%,98.65%for relative recovery(n=3). The limit of detection (LOD)(S/N=3) was about1.94ng, and the limits of quantification was0.97ng.Analytical linearity of TL-3was obtained for the method over the concentration range of0.36-3.28μg and the correlation coefficient r2=0.9997. The relative standard deviation (RSD) was1.02%for intra-day precision assay (n=6). The RSD of high concentration, median concentration and low concentration were0.68%,0.30%,1.05%for inter-day precision assay (n=5) respectively. The RSD of high concentration, median concentration and low concentration were101.86%,98.72%,100.28%for relative recovery(n=3). The limit of detection (LOD)(S/N=3) was about6.07ng, and the limits of quantification was2ng.Conclusions:TZT-9tablets can improve patient compliance. Through inspecting molding craft, safe and quality stability tablets were formulated. The content determination method in quality standard is reproducible, simple, rapid and can be used for the quality control of TZT-9tablets. |
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