ANTITUMOR CATALYTIC ANTIBODIES

化疗是临床治疗恶性肿瘤的有力武器， 癌不幸的是，大多数抗肿瘤药物与 严重的副作用，如严重的胃肠道和骨骼 骨髓毒性这一特点在很大程度上是由于 这些高毒性药物对靶点缺乏选择性 肿瘤细胞高于正常细胞。因此，无论是设计 靶向药物或有效的药物递送载体对于 推进癌症化疗。该建议旨在制定一项 新的和一般性的战略，为特定地点的交付， 依赖于设计和 针对该位点的靶向催化抗体（CAb）的优化- 抗肿瘤剂氨基甲酸酯前药的特异性释放。初始 模型研究将集中于油井的输送 表征的荧光剂5-氟尿苷（FUDR）和 阿糖胞苷（ara-C）。在此期间开发的方法 调查将适用于未来交付的 现有的和未来的化疗药物。这一目标 建议如下： L.设计并合成：a）芳香族氨基酸氨基甲酸酯 FUDR和ara-C的前药，和B）能够 产生CAbs， FUDR和ara-C的前药。 2.产生噬菌体融合蛋白： 来自用氨基磷酸酯半抗原免疫的小鼠的文库，和B） 来自幼稚外周B淋巴细胞的人抗体文库。 3.通过以下步骤从抗体噬菌体文库产生CAb：a） 用显色琼脂平板测定法鉴定活性催化剂， B）CAb的细菌表达，和c）动力学 表征纯化的CAb的功效。 4.为了构建能够结合 表面展示人癌细胞转铁蛋白受体， 体外 5.为了评价双特异性催化抗体在免疫调节中的能力， 与FUDR或ara-C的氨基甲酸酯前药组合以抑制 在体外的人癌的生长，其表面显示 转铁蛋白受体

Chemotherapy is a potent weapon in the clinical treatment of cancer. Unfortunately, most antitumor agents are associated with serious side effects such as severe gastrointestinal and bone marrow toxicity. This characteristic is due in large part to the lack of selectivity of these highly toxic drugs for the target tumor cells over normal cells. Therefore, the design of either targeted drugs or effective drug delivery vehicles is crucial to advancing cancer chemotherapy. This proposal seeks to develop a novel and general strategy for the site-specific delivery of antineoplastic therapeutics that will rely on the design and optimization of targeted catalytic antibodies (CAbs) for the site- specific release of carbamate prodrugs of antitumor agents. Initial model studies will concentrate on the delivery of the well characterized antineoplastic agents 5-fluorouridine (FUDR) and cytosine arabanoside (ara-C). Methods developed during these investigations will be applicable for the future delivery of existing and future chemotherapeutics. The objectives of this proposal are: l. To design and synthesize: a) aromatic amino acid carbamate prodrugs of FUDR and ara-C, and b) phosporamidate haptens capable of generating CAbs that will activate aromatic amino acid carbamate prodrugs of FUDR and ara-C. 2. To generate as bacteriophage fusion proteins: a) murine antibody libraries from mice immunized with phosphoramidate haptens, and b) human antibody libraries from naive peripheral B-lymphocytes. 3. To generate CAbs from an antibody bacteriophage library by: a) identifying active catalysts with a chromogenic agar plate assay, b) the bacterial expression of CAbs, and c) kinetically characterizing the efficacy of the purified CAbs. 4. To construct bispecific catalytic antibodies capable of binding the surface displayed transferrin receptor of human carcinomas in vitro. 5. To evaluate the ability of bispecific catalytic antibodies in combination with carbamate prodrugs of FUDR or ara-C to inhibit the growth of human carcinomas in vitro that surface display the transferrin receptor.