Delivery of Agents by Modulating Junctions with Zot

描述（由申请人提供）：强效大分子的鉴定正以惊人的速度增加，但由于理化限制，许多大分子无法有效递送。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，以增强细胞旁药物转运。我们最近已经证明，封闭小带毒素（Zot）Zot，一种45 kDa的蛋白质，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。该技术允许增强的细胞旁通量，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技术可以通过可逆地打开紧密的细胞连接，通过口服，血脑屏障和鼻腔递送增强药物递送。将努力实现以下具体目标。SAI。明确Zot主要结构域与跨上皮/内皮细胞旁通路调控之间的结构-功能关系。SA 2.研究Zot的剂量反应、药代动力学（PK）和急性毒性。我们将定义体内Zot（或衍生物）的完整剂量-反应曲线。SA 3.检查Zot增强亲水性药物（甘露醇、菊粉）和治疗性大分子（胰岛素、环孢菌素A、紫杉醇）口服生物利用度的能力。初步数据显示Zot增强治疗分子的口服吸收。SA 4.检查Zot增强最小转运CNS治疗性大分子的BBB递送的能力。Zot受体的配体Zonulin已在脑中发现，并且数据显示Zot增强治疗剂（多柔比星、菊粉、阿昔洛韦）的BBB递送。SA 5.为了检查Zot作为一种新型粘膜佐剂的能力，其增强大分子量治疗性大分子（例如，肽类，B的保护性抗原。炭疽病）。已经发现Zot在经鼻递送后显著增强破伤风毒素（TT）抗体的产生。针对SA 3-5提出的体内研究将评价Zot增强结构多样的治疗性大分子的全身水平的能力。Zot技术的潜在影响是显著和全面的，因为它可以应用于通过口服、BBB或鼻腔递送将多种大分子递送至其靶标。

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.