Dissertation > Excellent graduate degree dissertation topics show

Development of the Poly (Ethylene Glycol)-poly (Lactic Acid)-mitomycin C Micelle

Author: YueChangLai
Tutor: ZhangYingGe
School: PLA Military Academy of Medical Sciences
Course: Pharmacology
Keywords: mPEG-PLA MMC Antitumor Micelle Tissue injury
CLC: TQ460.1
Type: Master's thesis
Year: 2010
Downloads: 246
Quote: 0
Read: Download Dissertation

Abstract


This test is designed to prepare single-methoxy the base polyethylene glycol - polylactic acid - mitomycin Nanomicelles its pharmacodynamic evaluation. Monomethoxy polyethylene glycol - a polylactic acid block polymer (mPEG-PLA) was first prepared, and analyzed by infrared spectroscopy and H1NMR the polymer, to determine the structure of the polymer and then using their characterization; the acetone - the volatile prepared mPEG-PLA-MMC Nanomicelles, the Nanomicelles 0.45um millipore filter-sterilized filtrate collected mPEG-PLA-MMC micelles were freeze-dried solid. Micellar particle size and morphology using transmission electron microscopy, atomic force microscopy to characterize; utilize ultraviolet spectrophotometry mPEG-PLA-MMC Nanomicelles encapsulation efficiency and drug loading; the micellar nanocarrier particles effects burst release, release effect was measured by UV spectrophotometry; critical micelle concentration (CMC) was determined by fluorescence spectrophotometry; mPEG-PLA of-MMC Nanomicelles the release of research to take with PBS buffer, UV spectrophotometer method determination; mPEG-PLA block polymers using MTT assay in vitro, the pharmacological effects of MMC, the main experiments with cell human gastric adenocarcinoma BGC-823 cells, human hepatoma HepG2 cells; vivo test using a mouse sarcoma S180 hepatoma ascites tumor H22 model study the pharmacological effects Nanomicelles mPEG-PLA-MMC, compared to single use MMC, study the micelles drugs with enhanced targeting and pharmacological effects of MMC. A single-methoxy polyethylene glycol - poly-lactic acid block polymer (mPEG-PLA) Preparation and Characterization of monomethoxy polyethylene glycol and D, L-lactide as raw materials, through direct open ring The polymerization reaction is the MPEG-PLA block polymer was prepared. MPEG and D, L-lactide was placed in a vacuum oven at 70 ° C, dried 2H 0.1mPa under pressure. The toluene was purified by distillation to spare. The mPEG and D, L-propan-lactide sealed into a three-necked bottle in accordance with a certain quality, stannous octoate as the catalyst, nitrogen protection under conditions of 140 ° C under reflux 8H end of the reaction, monomethoxy polyethylene glycol alcohol - polylactic acid block polymer (mPEG-PLA) and the crude product purified to obtain a more pure product, the color is white or milky white. The block polymer characterization is mainly taken the infrared spectroscopy and H1NMR of analysis. Can be determined by the analysis of the samples prepared as monomethoxy polyethylene glycol - polylactic acid block polymer (mPEG-PLA). Second, the determination of the critical micelle concentration (CMC) as a probe to a fluorescent substance pyrene MPEG-PLA block polymer of the CMC values ??can be determined, as the solution MPEG-PLA copolymer concentration increases the excitation light intensity, pyrene increases, a red shift of the excitation wavelength. When the copolymer concentration above the CMC value, pyrene into the micelles, the ratio of I338/1333 change significantly, the CMC value of the copolymer can be determined through this significant change point. MPEG-PLA solution was prepared containing different concentrations of a certain amount of pyrene measured under the fluorescence spectrophotometer, and the results show that the CMC value of 1.9mg / L. , MPEG-PLA-MMC Preparation and characterization of the micelle 1 Preparation of mPEG-PLA-MMC micelles: MPEG-PLA block polymer with the MMC in accordance with the different quality than the miscibility in the certain volume of the acetone solution, constant stirring so that all the solid dissolved. Dark conditions in a fume hood, the solution in accordance with the speed of 1 drop / s was added dropwise to contain a certain amount of the triple-distilled water in a beaker, the beaker contains a magnetic stirring bar, mixed dropwise with a solution of tri-distilled water, ventilation under conditions the acetone as volatilization completely as possible. After a certain time, remove the solution, it is transferred into the prior handling of the dialysis bag (molecular weight cutoff of the dialysis bag ≥ 5000d), dialyzed in a thermostat water bath oscillator purpose MMC liberated, while micellar preserved. The MMC content of the dialysis bag was measured to determine whether the dialysis completely. Wait until the end of dialysis, the dialysis bag micelle transfer to sterilized 0.45um millipore filter sterilization. The filtrate is collected by autoclaving glassware, pre-frozen at -20 ℃. The end of the pre-frozen, then transferred to a freeze drier, the micelles prepared freeze-dried solid. Prepared mPEG-PLA-MMC Nanomicelles kept sealed, dark, under normal temperature conditions. 2, the particle size of the micelles, morphological characterization 2.1, transmission electron microscopy method transmission electron microscopy (TEM) results showed: dry micelles as spherical particles can form a core-shell structure. Intermediate density greater hydrophobicity nuclear shiny i.e. PLA, outer periphery of the touch layer the hydrophilicity of MPEG. Blank micelles particle diameter of about 50nm, and the particle diameter of drug-loaded micelles is between 80 ~ 100nm, the particle size distribution more uniform. 2.2, atomic force microscopy preparation micellar solution of a certain concentration, the ultrasound it as much as possible separation to reduce agglomeration. After the ultrasound solution was added dropwise to freshly cleaved flat mica surface, fixed 5min with filter paper, the solution sucking, and placed in the air dried completely. Dry completely after the samples were placed in an atomic force microscope observation determination. The results showed: micelle particle size is consistent with transmission electron microscopy results. 2.3, infrared spectroscopy using KBr pellets infrared spectrometer analysis the blank micelles of mPEG-PLA-MMC nano-micelles, mPEG-PLA and mPEG-PLA blank micelles with MMC physical mixture of three different samples. Comparative infrared spectra, it is determined that the MMC does exist in the prepared complexes. 2.4, H1NMR method take the amount of sample, which was dissolved to CH2DCCl3, the solution was transferred to the NMR tube, 400MHz under conditions analyzed. Magnetic resonance images, by comparison mPEG-PLA-MMC nano-micelles, mPEG-PLA blank micelles and mPEG-PLA blank micelles with MMC physical mixing determine MMC exist in mPEG-PLA block copolymer, prepared need mPEG-PLA-MMC nano-micelles. Determination of mPEG-PLA-MMC micelle drug loading and encapsulation efficiency and sustained release of the process of Preparation of mPEG-PLA-MMC micelles, collecting the solution outside the dialysis bag, wherein the free MMC absorbance was measured into the MMC standard curves A = 0.0042 0.0607 * C (R = 1.0000, P lt; 0.0001), can be obtained free of MMC's quality. Minus the total MMC free MMC's quality, the quality of the micelles, mPEG-PLA-MMC, MMC can be obtained using the following formula seeking drug-loaded micelles drug loading and encapsulation efficiency. Calculated when mPEG-PLA block polymer with MMC quality ratio is 10:3, the drug-loaded micelles drug loading and encapsulation efficiency is better, up to 15% drug loading, encapsulation efficiency of up to 52 %. Said certain amount of drug-loaded micelles, which was dissolved in a certain volume of PBS, transferred to a dialysis bag (molecular weight cutoff of ≥ 5000d) by the pre-processing to be sealed and placed in PBS beaker is equipped with a large number of The beaker was placed in a thermostat water bath oscillator, to set the temperature and speed, at a predetermined time point in time replacement of a fixed amount of fresh PBS liquid, can not be detected until the use of UV-method to the MMC. The content of accumulated with the mapping of the corresponding point in time, you can get the mPEG-PLA-MMC micelles burst release, sustained-release effect diagram. Experimental results show that the drug-loaded micelles is not obvious burst release, sustained-release effect release time of 18 ± 1d. , MPEG-PLA studies of MMC in vitro anti-cancer cells in vitro tests designed to examine from the perspective of in vitro mPEG-PLA pharmacological effects of MMC. MTT assay was used to prove that by measuring the inhibition rate of mPEG-PLA whether it will affect the MMC killer cells. BGC-823 cells and Hepg2 of cells used in the experiment as experimental subjects. Experimental results show that mPEG-PLA-MMC opposite at lower doses Nanomicelles anti-tumor activity in vitro mPEG-PLA block polymer enhanced role in the activity of the MMC, lower in the high dose (200μg/ml above) there was no significant difference. , The generation of the results of this experiment is reasonable taking into account the effect of sustained release of the mPEG-PLA-MMC Nanomicelles low dose because of mPEG-PLA-MMC Nanomicelles release of MMC less and therefore inhibition lower, high release more doses of MMC, MMC reaches a certain amount, the inhibition of cell is relatively stable, so the difference between the two is not significant. Six, mPEG-PLA MMC vivo anti-tumor effect and toxicity studies in vivo test mice sarcoma S180 model tumor inhibition rate and to extend the life of the mouse hepatoma ascites tumor H22 model rate observed mPEG-PLA block polymerization was in the body of the MMC synergies. Toxicity study BABL / C mice, a series of dose, observed tissue injury. 1 mouse sarcoma S180 model to mice sarcoma S180 test model mPEG-PLA block polymer can significantly increase the MMC in vivo anti-tumor effect. mPEG-PLA-MMC Nanomicelles dose for 2,6,18 mg / kg inhibition of tumor was 31.23%, 51.78%, 66.80%, and the MMC dose of 1mg/kg of 34.78% inhibition of tumor. mPEG-PLA-MMC Nanomicelles MMC's content is about 1/6, mPEG-PLA-MMC Nanomicelles for 6mg/kg when single use MMC1mg/kg dose quite. The data show that: mPEG-PLA the-MMC Nanomicelles dose of 2mg/kg when single-use MMC dose 1mg/kg considerable inhibition of tumor in the middle and high dose inhibition of tumor significantly increase was statistically significant (P lt; 0.001). Mouse hepatoma H22 model ascites tumor ascites tumor in mouse hepatoma H22 test model mPEG-PLA-MMC Nanomicelles group can significantly prolong the survival time of tumor-bearing mice, and extend the life extension rate. mPEG-PLA-MMC Nanomicelles dose 2,6,18 mg / kg life extension rate of 76.6%, 171.0% and 206.5%, respectively, the MMC dose 1mg/kg life extension rate of 123.4%. Data show that: mPEG-PLA the-MMC Nanomicelles dose of 2mg/kg when single-use MMC dose 1mg/kg low rate of life extension, and the difference was statistically significant (P lt; 0.01) between dose of 6,18 mg / kg when single-use MMC dose of 1mg/kg effect on prolonging the lives of mice increased significantly, with a statistically significant (P lt; 0.01). 3 local tissue damage experimental results showed that: mPEG-PLA-MMC Nanomicelles-MMC tissue damage can be significantly reduced. mPEG-PLA-MMC Nanomicelles group MMC group at the time of tissue damage than late, the incidence of low, smaller damage. lowest dose of mPEG-PLA-MMC Nanomicelles group injury 0.08mg MMC experimental group was 0.02 mg; less tissue damage area mPEG-PLA-MMC the Nanomicelles group relative to the same dose of MMC, a statistically significant difference (P lt; 0.05 or P lt; 0.01).

Related Dissertations

  1. Design, Synthesis and Biological Evaluation of Novel Podophyllotoxin Derivatives as Antitumor Agents,R284
  2. Research on Biological Modification and Protection Activity of R-Phycoerythin from Porphyra Yezoensis,TS201.21
  3. Novel Imidazophenanthroline Derivatives: Design, Synthesis and Evaluation of Biological Activity,TQ460.1
  4. Expression, Purification and Characterization of Trucated Type Recombinant Buckwheat Trypsin Inhibitor,R284.1
  5. Antitumor Effect and Expression of the Pro-apoptotic Bcl-2 Family Member N of Elemene in Mice Bearing Hepatocarcinoma H22 Cells-derived Tumor,R285.5
  6. Establishment of Subcutaneous Transplanted Tumor Model of Skin Adenocarcinoma in Balb/c Mice,R739.5
  7. 1, Bcl-2 small molecule inhibitors of protein synthesis and antitumor activity of two hydroxytyrosol new methods of synthesis,R96
  8. Design, Synthesis and Study of Antitumor Activity of Novel Small Molecular Inhibitors of PKB,R914
  9. Preparation of Doxorubicin Hydrochloride Nanoliposome and Its in Vitro Antitumor Activity,R96
  10. Study on Extraction and Antioxidant Activity, Anti-tumor Activity and Anti-bacterial Activity of Flavonoid from Halianths Tuber Leaves,S632.9
  11. Preliminary Study on Anti-tumor Effect of Sinapine on H22 Mice Hepatocellular Carcinoma and Its Mechanisms,R735.7
  12. Association Analysis on MMP-3 Polymorphism and Breast Cancer and Anti-tumor Mechanism Study on Long-quan Compound Traditional Chinese Medicine,R737.9
  13. The Study of Evaluating the Effect and Safety of Mitomycin C and Tetrandrine on the Prevention of Posterior Capsular Opacification,R779.66
  14. Preparation of Simvastatins-loaded MePEG-PLA Nanoparticles and Effects of Simvastatins-loaded MePEG-PLA Nanoparticles on Osteoblasts,R965
  15. Purification of Antitumor Protein from Trachyrhamphus Serratus and Investigation of It’s in Vitro Avtivity,R96
  16. Friction and corrosion behavior of complex metal alloys ( CMA ) reinforced metal matrix composites ( MMC ),TB331
  17. Preparation and in Vitro Antioxidant and Antitumor Activities of R-Phycoerythrin from Porphyra Yezoensis,TS201.21
  18. Expression, Purification and Antitumor Activities of a New Recombinant Human Interferon-λ1 (IL-29),R96
  19. Extraction of Total Flavonoids from Tartary Buckwheat and Its Anti-tumor Activity in Vitro,S517
  20. Screening of Antitumor Fractions of Solanum Lyratum and Preliminary Study on Its Mechanism,R285
  21. Synthesis and Characterization of a Novel Degradabable Dendrimer and Preparation of Its Anti-tumor Prodrug,R914

CLC: > Industrial Technology > Chemical Industry > Pharmaceutical chemical industry > General issues > Basic theory
© 2012 www.DissertationTopic.Net  Mobile