Modified Human Anti-a(3) IV Antibodies for Drug Delivery in Nephritis

描述(申请人提供)：限制人类疾病的长期目标。为了实现这一目标，我们将利用以下优势：13(IV)胶原蛋白(13(IV))的限制性及其与肾脏的可用表位；体内可与13(IV)肾小球结合的一组独特的人类单抗(MAb)的可用性，以及我们在抗体工程、蛋白质表达和肾炎实验模型方面的经验，以创建体内与肾脏结合的人类融合蛋白。实验方法将涉及对人m抗13(IV)抗体的修饰，从而保持它们与13(IV)的抗原结合，从而使它们与小鼠体内的GBM结合。然而，它们的Fc区域将被改变，因此它们将被构建成具有向肾小球传递抗炎或抗纤维化信号的潜力，而不是促进炎症。最初，这将涉及到候选人源抗13(IV)微体的体外筛选、表达和纯化。随后，将在正常小鼠和肾炎小鼠中进行进一步的候选筛选和优化，以确定理想的试剂输送到肾小球，长期目标是输送疾病改良剂。这些技术以及上述试剂小组将被用于实现以下具体目标：1：生产和鉴定带有连接子的重组人抗13(IV)抗体片段；2：评估在健康和疾病期间，人抗体片段将试剂运送到体内肾脏的能力。一旦条件得到优化，这些试剂将被用于未来的研究，以检验这样的假设，即修改后的人抗13(IV)抗体将有效地将药物输送到肾脏，以限制局部炎症，而不会产生全身副作用。如果成功，这种方法和技术的一个主要优势是它将适用于所有形式的肾小球肾炎。此外，由于这些试剂来自人类抗体序列，它们有可能被直接评估为治疗人类肾小球肾炎，只需有限的修改。公共卫生相关性：这些研究的总体目标是开发更有效、毒性更低的肾炎治疗方法。为此，与肾脏结合的人类抗体将被修饰，因此它们有可能定位于肾脏，并提供抗炎和抗纤维化药物，以改变肾炎的病程。虽然拟议的研究将在老鼠身上进行，但由于这些试剂将以人类序列为基础，它们将有可能在未来直接应用于各种形式的人类肾小球肾炎。

DESCRIPTION (provided by applicant): The long-term aim of to limit disease in man. Toward this goal, we will take advantage of: the restricted nature of 13(IV) collagen (13(IV) and its available epitopes with the kidney; the availability of a unique panel of human monoclonal antibodies (mAb) that bind to 13(IV) within glomeruli in vivo, and our experience in Ab engineering, protein expression, and experimental models of nephritis, to create human fusion proteins that bind to the kidney, in vivo. The experimental approach will involve modification of human m anti-13(IV) Ab, whereby their antigen binding for 13(IV) is maintained, so that they bind to the GBM in vivo, in mice. However their Fc regions will be altered, so that instead of promoting inflammation, they will constructed so that they have the potential to deliver either anti-inflammatory or anti-fibrotic signals to glomeruli. Initially, this will involve, selection expression and purification of candidate human m anti-13(IV) minibodies, in vitro. Subsequently, further selection and optimization of candidates will be carried out in normal, and then in nephritic mice, to identify the ideal reagents for delivery to glomeruli, with the long-term goal to deliver disease modifying agents. These technologies, along with the aforementioned panel of reagents, will be used to pursue the following Specific Aims: 1: To produce and characterize recombinant human anti-13(IV)Ab fragments with linkers, and 2: To evaluate the capacity of the human Ab fragments to deliver reagents to the kidney, in vivo, during health and disease. Once conditions are optimized, the reagents will be used in future studies to test the hypothesis that modified human anti-13 (IV) Ab will efficiently deliver drugs to the kidney to limit local inflammation, without systemic side effects. If successful, a major advantage of this approach and technology is that it will have application to all forms of glomerulonephritis. Furthermore, since the reagents are derived from human Ab sequences, they have the potential of being directly evaluated to treat human glomerulonephritis, with limited modification. PUBLIC HEALTH RELEVANCE: The overall goal of these studies is develop more effective, less toxic therapy for glomerulonephritis. For this purpose, human antibodies that bind to the kidney will be modified, so they have the potential to localize in the kidney and deliver anti- inflammatory and anti-fibrotic agents, to modify the course of nephritis. Although the proposed studies will be performed in mice, since the reagents will be based on human sequences, they will have the potential to be directly applied to various forms of human glomerulonephritis in the future.