ENLARGED PATHWAYS FOR TRANSDERMAL PROTEIN DELIVERY

描述：(改编自申请人的摘要)-这项建议的目标是建立大分子(例如，肽、蛋白质和核酸)的经皮给药方法，具有极好的控制和低成本的潜力。申请者将研究通过皮肤角质层(SC)的扩大的经皮透皮水通道，这是在“高电压”(HV)脉冲(Ukin大于或等于50V)电产生初级水后的二次过程中产生的。目标。由于涉及的参数很多，其目的是在体外广泛研究和优化通路扩大，然后是体内动物试验，包括通路创建、大分子转运和副作用的初步评估。意义重大。这些目标的成功实现将为基于紧凑、低成本的电化学技术的经皮给药奠定基础。这项设想的技术将提供电子控制的快速水溶性分子，范围从传统药物到体内半衰期短的大分子。因此，注射器的许多用途都可以被取代。以前的工作。非优化方案表明，(1)纯粹的物理刺激可能会产生更大的通路，(2)无毒的通路扩大分子会产生更大的通路，以潜在的治疗水平(每厘米每小时100微克/平方厘米)运输蛋白质。根据未来设备的要求，传输似乎集中在占1平方厘米皮肤面积不到10%的小皮肤部位(局部传输区域；“LTRS”)。使用微电极和微孔的原型电极/储液器设备在体内提供无痛脉冲。申请人和其他人的实验和局部电场的理论建模都表明，副作用应该是最小的。在一个案例中，抗体分子(免疫球蛋白每摩尔150,000克)被运输。方法：研究方法。申请者将强调多参数扩大途径创建方案的体外优化，使用人、无毛大鼠和猪的皮肤，然后进行体内评估。他们将使用皮肤电阻和荧光分子通量测量并结合图像分析来表征包含更大路径的LTRs。将对局域场、加热、路径创建和传输进行理论分析，这对于优化扩大的路径是必不可少的。

DESCRIPTION: (Adapted from the applicant's abstract) - The goal of this proposal is to establish methods for transdermal delivery of macromolecules, (e.g., peptides, proteins, and nucleic acids), with the potential for excellent control and low cost. The applicants will study enlarged transdermal aqueous pathways through the skin's stratum corneum (SC), that are created by secondary processes occurring after electrical creation of primary aqueous by "high- voltage" (HV) pulses (Uskin greater than or equal to 50V). Aims. Because of the many parameters involved, the aims are to study extensively and optimize pathway enlargement in vitro, followed by in vivo animal studies involving pathway creation, macromolecule transport, and initial evaluation of side effects. Significance. Successful pursuit of the Aims will establish a basis for transdermal drug delivery based on a compact, low-cost electrochemical technology. The envisioned technology will provide electronically controlled rapid delivery of water-soluble molecules ranging from conventional pharmaceuticals to macromolecules with short in vivo half lives. Thus, many of the uses of syringes could be replaced. Previous Work. Nonoptimized protocols show that (1) purely physical stimuli may create enlarged pathways, and (2) a nontoxic pathway-enlarging molecule created enlarged pathways which transported proteins at potentially therapeutic levels (100 micrograms per hour per centimeter squared). As required for future devices, transport appears to be concentrated at small skin sites (local transport regions; "LTRs") which occupy less than 10% of a 1 square centimeter area of skin. Prototype electrode/reservoir devices using microelectrodes and microholes provide painless pulsing in vivo. Both experiments (of the applicants and others) and theoretical modeling of localized electric fields show that side effects should be minimal. In one case, antibody molecules (IGG at 150,000 g per mole) were transported. Methods. The applicants will emphasize in vitro optimization of multiparametric enlarged pathway-creation protocols, using human, hairless rat, and pig skin, followed by in vivo assessment. They will use skin electrical resistance and fluorescent molecule flux measurements with image analysis to characterize the LTRs containing enlarged pathways. Theoretical analysis of localized fields, heating, pathway creation, and transport will be carried out, and is essential to optimizing enlarged pathways.