Therapeutic Targeting Agents for Bladder Cancer

描述（由申请人提供）：本项目的目的是开发用于治疗膀胱癌的新型细胞选择性治疗靶向药物。利用已经开发的两种非重叠噬菌体展示衍生的膀胱癌特异性结合肽（BCSBP），将在这些BCSBP与活性化疗药物（Epiradine）和两种生物蛋白（细胞因子IL-2和IL-12）之间产生缀合物。这些BCSBP的结合和功能活性将针对先前显示与多种人膀胱癌细胞系共享这些BCSBP的相同结合特征的小鼠膀胱癌细胞系进行优化。长期目标是开发必要的方法，将这些靶向基序转化为针对人类膀胱癌的临床应用免疫学。为了实现这些目标，将实现两个具体目标： 具体目标1：将BCSBPs与化疗药物Epiradine化学偶联，并评估偶联物靶向膀胱癌细胞的细胞毒性。将通过化学偶联过程制备缀合物，该化学偶联过程探索位置效应、靶向基序的数量和间隔区的使用以实现最大结合特异性、亲和力和功能性细胞毒性活性。 具体目标2：将BCSBP与Th 1细胞因子IL-2和IL-12基因偶联，并评估这些融合蛋白对靶向膀胱癌细胞的结合和生物活性。这些融合蛋白将使用原核或昆虫表达系统制备。将使用GFP缀合物原型优化结合特异性。将使用完善的生物测定法测试IL-2和IL-12单独和作为协同剂的生物活性。本研究的成功完成将导致在动物膀胱癌模型中使用这些新型治疗性缀合物的未来临床前研究，这可能进一步导致将该策略快速转化为临床上有用的人膀胱癌治疗方式。

DESCRIPTION (provided by applicant): The purpose of this project is to develop novel cell-selective therapeutic targeting agents for treating bladder cancer. Making use of two non-overlapping phage-display derived bladder cancer-specific binding peptides (BCSBPs) already developed, conjugates will be created between these BCSBPs and both an active chemotherapeutic drug (Epirubicin) and two biologic proteins (cytokines IL-2 and IL-12). Binding and functional activity of these BCSBPs will be optimized against a mouse bladder cancer cell line previously shown to share the same binding characteristics for these BCSBPs as multiple human bladder cancer cell lines. The long-term goal is to develop the necessary methodology for translating these targeting motifs into clinical use immunotherapeutic against human bladder cancer. Towards achieving these goals, two specific aims will be undertaken: Specific Aim 1: To chemically conjugate BCSBPs with the chemotherapeutic drug Epirubicin, and to assess cytotoxicity of the conjugates in targeting bladder cancer cells. The conjugates will be made through a chemical coupling process that explores positional effects, number of targeting motifs and use of spacers to achieve maximal binding specificity, affinity, and functional cytotoxic activity. Specific aim 2: To genetically conjugate BCSBPs with Thl cytokines IL-2 and IL-12, and to assess the binding and biological activities of these fusion proteins upon targeted bladder cancer cells. These fusion proteins will be made using prokaryotic or insect expression systems. Binding specificity will be optimized using a GFP-conjugate prototype. Biological activity for IL-2 and IL-12, alone and as synergistic agents will be tested using well-established bioassays. Successful completion of this study will result in future pre-clinical studies using these novel therapeutic conjugates in animal bladder cancer models that may further lead to a quick translation of this strategy into a clinically useful treatment modality for human bladder cancer.