Delivery of Agents by Modulating Junctions with Zot

描述（由申请人提供）：有效大分子的鉴定正以惊人的速度增长，然而由于物理化学的限制，许多大分子不能有效地递送。一种可行的药物递送方法可能是利用细胞间紧密连接的生理调节，以增强细胞旁药物运输。我们最近证明，封闭带毒素（Zot） Zot是一种45kDa的蛋白，通过模拟负责调节细胞间紧密连接的真核类似物来发挥其渗透作用。这种技术允许增强细胞旁通量，并具有有效递送低生物利用度治疗大分子的潜力。因此，我们的假设是Zot技术可以通过口服、血脑屏障和鼻腔给药，可逆地打开紧密的细胞连接，从而增强药物的传递。将实现下列具体目标。赛。定义Zot主要结构域与经上皮/内皮细胞旁通路调控之间的结构-功能关系。SA2。目的：测定Zot的剂量反应、药代动力学（PK）和急性毒性。我们将定义Zot（或衍生物）在体内的完整剂量-反应谱。SA3。研究Zot提高亲水性制剂（甘露醇、菊糖）和治疗性大分子（胰岛素、环孢素A、紫杉醇）口服生物利用度的能力。初步数据显示Zot能增强口服治疗分子的吸收。SA4。研究Zot增强中枢神经系统最小转运治疗性大分子血脑屏障递送的能力。Zonulin是Zot受体的配体，已经在大脑中发现，数据表明Zot增强了治疗药物（阿霉素、菊粉、无环鸟苷）的血脑屏障递送。SA5。研究Zot作为一种新型粘膜佐剂的能力，提高大分子量治疗性大分子（如肽、对炭疽杆菌的保护性抗原）经鼻给药后的全身生物利用度。经鼻给药后发现Zot可显著提高破伤风毒素（TT）抗体的产生。SA 3-5的体内研究将评估Zot提高结构多样化治疗大分子的系统水平的能力。Zot技术的潜在影响是重要和全面的，因为它可以应用于通过口服、血脑屏障或鼻腔给药将多种大分子递送到它们的目标。

DESCRIPTION (provided by applicant): The identification of potent macromolecules is increasing at a staggering rate, however many are not delivered effectively due to physiochemical limitations. A viable approach to drug delivery may be to exploit the physiological regulation of intercellular tight junctions, to enhance paracellular drug transport. We have recently demonstrated that Zonula Occludens Toxin (Zot) Zot, a 45kDa protein, exerts its permeating effect by mimicking a eukaryotic analogue in charge of modulation of intercellular tight junctions This technology allows for enhanced paracellular flux and has the potential to effectively deliver low bioavailable therapeutic macromolecules. Thus our hypothesis is that the Zot technology can enhance drug delivery by reversibly opening tight cellular junctions via oral, BBB and nasal delivery. The following specific aims will be pursued. SAI. To define the structure-function relationships between the major domains of Zot and regulation of the transepithelial/endothelial paracellular pathway. SA2. To determine the dose response, pharmacokinetics (PK) and acute toxicity of Zot. We will define the complete dose-response profile of the Zot (or derivative) in vivo. SA3. To examine the ability of Zot to enhance the oral bioavailability of the hydrophilic agents (mannitol, inulin) and therapeutic macromolecules (insulin, cyclosporin A, paclitaxel). Preliminary data displays that Zot enhances oral absorption of therapeutic molecules. SA4. To examine the ability of Zot to enhance the BBB delivery of the minimally transported CNS therapeutic macromolecules. Zonulin, the ligand for the Zot receptor has been found in the brain and data has shown that Zot enhances the BBB delivery of therapeutic agents (doxorubicin, inulin, acyclovir). SA5. To examine the ability of Zot to act as a novel mucosal adjuvant that enhances the systemic bioavailability of large molecular weight therapeutic macromolecules (e.g., peptides, Protective antigen to B. anthracis) after nasal delivery. Zot has been found to significantly enhance the production of tetanus toxin (TT) antibodies after nasal delivery. The in vivo studies proposed for SA 3-5 will evaluate the ability of Zot to enhance the systemic levels of structurally diverse therapeutic macromolecules. The potential impact of the Zot technology is significant and comprehensive, since it may be applied to delivering a diversified range of macromolecules to their targets via oral, BBB or nasal delivery.