Therapeutic Targeting Agents for Bladder Cancer

描述(申请人提供)：本项目的目的是开发治疗膀胱癌的新型细胞选择性靶向药物。利用已经开发的两个不重叠的噬菌体展示衍生的膀胱癌特异性结合肽(BCSBPs)，这些BCSBPs将与一种有效的化疗药物(表阿霉素)和两种生物蛋白(细胞因子IL-2和IL-12)产生偶联物。这些BCSBPs的结合和功能活性将针对先前显示的与多种人类膀胱癌细胞株具有相同结合特性的小鼠膀胱癌细胞株进行优化。长期目标是开发必要的方法学，将这些靶向基序转化为临床使用，用于治疗人类膀胱癌。为实现这些目标，将实现两个具体目标： 具体目标1：将BCSBPs与化疗药物表阿霉素进行化学偶联，并评价其靶向膀胱癌细胞的细胞毒性。这些偶联物将通过化学偶联过程来探索位置效应、靶向基序的数量和间隔物的使用，以实现最大的结合特异性、亲和力和功能性细胞毒活性。 特定目的2：将BCSBPs与Th1细胞因子IL-2和IL-12进行基因偶联，并评价这些融合蛋白对靶向膀胱癌细胞的结合和生物学活性。这些融合蛋白将使用原核或昆虫表达系统来制造。结合特异性将使用GFP结合原型进行优化。IL-2和IL-12的生物活性将使用成熟的生物测定方法进行测试，无论是单独的还是作为协同作用的。这项研究的成功完成将导致未来在动物膀胱癌模型中使用这些新型治疗结合物进行临床前研究，从而进一步将这一策略迅速转化为临床上有用的人类膀胱癌治疗方法。

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.