Development of dry gene powder for lung and evaluation of its efficacy

Recent y precipitation of powders with supercritical carbon dioxide-（SCF） has been attracting much attention as a method to produce tary partides with high functionality We have already reported that the gene powders grew cl by the SCF prep bad improved stability and iir aced gene expression in lungs after intratracheal insufflation in mice。在目前的研究中，我们准备了利用SCF过程来释放甲状腺素干扰素β （DNA）的动力，以消除淀粉样淀粉样蛋白酶模型总线中的治疗方法：pCMV-MuBβ a等离子体DNA Cabling Murine intern β的解决方案，而chitosan，一种非病毒载体被转移到SCF中，作为乙醇的一种修饰物，以准备它们。甘露醇被用作一种粉末车。要稳定一个长金属，CT 26，小鼠结肠癌细胞，被植入小鼠尾血管。DNA粉末或解决方案是管理上的非直接或间接地测试肺部重量、数量的变位节点和与时间有关的存活率。The ge（ge） ... More 在DNA管理局内没有表达后，在正常和癌症组织中被剥夺，而在其他机构中没有表达。DNA动力抑制了体重中的增加和数量的节点，并使老鼠的存活率比DNA解决方案少得多。内部管理比内部管理更有效，而不是内部管理。这些发现建议使用SCF过程在Murine lung metastasis模型中具有高治疗潜力的发现。下一项研究审查了用超临界碳二氧化物（CO2）的视点的DNA和体内英寸潜力准备的基因的稳定性。一种类似的chitosan-pCMV-Luc复合体解决方案含有甘露醇，被注入到超临界二氧化碳/乙醇的流中，以补充基因粉末。获得的宝石粉和基因解决方案在稳定室放置在25或40 ° C的时间为4周。通过电泳测试的基因的完整性和转染潜力，以及在Mice的超临界二氧化碳过程中进行的范脉冲转染研究中的降解，用制造过程中的侧DNA进行降解;如何，在动力人中回收的超coiled和开放的循环DNA中降解在存储过程中的降解比解决方案要慢很多。在补充中，能量比能量更有可能容纳相同DNA数量的解决方案。Chitosan对DNA在解决方案中稳定性的影响在解决方案中并不明显，但它提高了DNA在生产和储存中的稳定性。因此，一个带有载体的基因粉末是一种准备好使用脉冲性疾病的抑制疗法的提议的配方。Less（低）

Recent y precipitation of powders with supercritical carbon dioxide- (SCF) has been attracting much attention as a method to produce tarry partides with high functionality We have already reported that the gene powders grew cl by the SCF prep bad improved stability and iir aced gene expression in lungs after intratracheal insufflation in mice. In the present study, we prepared chitosan-interferon β (DNA) powders by the SCF process to exannne the therapeutics of there in murinelung metastasis modelAn bus :solution of pCMV-MuBβ a plasmid DNA cabling murine intern β, and chitosan, a nonviral vector was dispersed in SCF with ethanol as a modifier to precipitate them. Mannitol was used as a powder vehicle. To establish a lung metasitatis, CT26, mouse colon carcinoma cells, were injected intravenously into mouse tail vein. The DNA powder or solution was administered intratracheally or intravenously to examine the lung weighty, number of metastatic nodules, and survival rate with time. The ge … More ne expression after intratracheal administration of DNA was abserved in normal and cancer tissue in the lung, while no expression was observed in the other organs. The DNA powders suppressed the increase in lung weight and number of nodules and prolonged the survival of the mice with smaller dose of DNA than DNA solutions. Intratracheal administration was more effective than intravenous administration. These findings suggested that the DNA powders prepared by the SCF processs had high therapeutic potential in murine lung metastasis model.The next study examined the stability of a gene in powers prepared with supercritical carbon dioxide (CO2) from the viewpoints of the ternary structure of DNA and in vivo inch potential. An aqueous chitosan-pCMV-Luc complex solution containing mannitol was injected into the stream of a supercritical CO2/ethanol admixture to precipitate a gene powder. The obtained gem powders and gene solutions were placed in stability chambers at 25 or 40゜C for 4 weeks. The integrity, and transfection potency of the gene were examined by electrophoresis and in van pulmonary transfection study in mice The supercritical CO2 process decreased the sided DNA doting the manufacturing process; however, the decrease in the remaining supercoiled and open circular DNA in the powders during storage was much slower than that in solutions. In addition, the powders had W war w on potency than the solutions containing the same amount of DNA. The effect of chitosan on the stability of DNA in solutions was not obvious in the solutions but it improved the stability of DNA in powders du manufacturing and storage. Thus, a gene powder with a vector is a promising formulation for a ready-to use inhalation therapy of pulmonary diseases. Less