Molecular Requirements for Recombinant Cytotoxins Efficacy

重组细胞毒素功效的分子要求

基本信息

批准号：
7163020
负责人：
Waldemar Debinski
金额：
$ 24.38万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-12-28 至 2010-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7163020
关键词：
Affect Affinity Agonist Appendix Bacterial Toxins Binding Binding Sites Biological Brain Neoplasms Cell membrane Cells Cellular biology Chimeric Proteins Clinical Clinical Trials Condition Cytotoxin Decision Making Development Diphtheria Toxin Effectiveness Endopeptidases Engineering Enzymes Exhibits Exotoxins Family Future Generations Glioblastoma Glioma Helix (Snails)Homologous Gene Hypoxia IL4 gene In Situ In Vitro Interleukin-13 Interleukin-4 Knowledge Laboratories Ligands Link Malignant Glioma Malignant Neoplasms Measurable Mediating Membrane Proteins Modeling Molecular Molecular Target Mutate Neuraxis Normal tissue morphology Numbers Organ Patients Peptide Hydrolases Phase Phase III Clinical Trials Physiological Play Proteins Pseudomonas Pseudomonas aeruginosa toxA protein Recombinants Regulation Research Personnel Role Site Solid Neoplasm Solutions Specificity Structure Structure-Activity Relationship Targeted Toxins Testicular Neoplasms Testing Toxic effect Toxin Tumor Antigens United States Food and Drug Administration alpha helix base cancer cell cancer therapy cell killing cytokine cytotoxic design ear helix experience in vivo information gathering interest interleukin-13 receptor mutant neoplastic cell novel pre-clinical programs receptor research clinical testing response transcription factor tumor

项目摘要

DESCRIPTION (provided by applicant): A restricted receptor (R) for interleukin 13 (IL13) was found in a vast majority of high-grade gliomas (HGG) patients. The HGG-associated receptor for IL13 was identified molecularly to be the IL13R(2, and it belongs to a family of tumor antigens, termed cancer/testis tumor antigens (CTA). CTA provide high specificity for molecular targeting/recognition of cancer. Furthermore, IL13-based bacterial toxin, Pseudomonas exotoxin A (PE)-containing recombinant cytotoxins were documented to be the potent anti-glioma agents in pre-clinical evaluation, and the first generation of these cytotoxins is in Phase III clinical trials. The structure-function relationship analysis of IL13 and its recently revealed solution structure documented that alpha-helix D appears to play pivotal role in the binding of IL13 to its HGG-associated receptor. It is proposed to generate novel highly specific and highly efficacious IL13-based cytotoxins that will incorporate (i) new knowledge on IL13 structure-function relationship, (ii) new information on glioma cell biology, and (iii) previous experience with the use of cytotoxins in a clinical setting. Thus, these new cytotoxins will be composed of genetically engineered forms of IL13 and a derivative of another bacterial toxin, Diphtheria toxin, DT390. The principal idea behind the design of these cytotoxins is to produce novel cytotoxins that allow the binding region of IL13, the D-helix, to be freely available to the HGG-associated receptor for IL13 and to eliminate the site on the cytokine that interacts with its normal physiological receptor that is expressed in many vital organs, including the central nervous system. IL13 will be engineered to have the DT toxin moiety remote form its binding site to the IL13R(2, which cannot be fully achieved with PE. Moreover, IL13 alpha-helix D mutants, super agonists of the IL13R(2, have been identified and they will be used in the design of optimized cytotoxins. The number of the restricted binding sites for the cytotoxins and an intracellular protease activating bacterial toxins (furin) play major roles in the cytotoxins' tumor cell killing. Thus, the regulation of their expression levels will be examined and the expression levels correlated with the novel IL13-DT cytotoxins efficacy. The novel DT390-IL13 mutant-based cytotoxins will be tested in vitro and in vivo in order to demonstrate how the changes in the molecular design in the context of biological expression of the target and toxin's activating moiety are reflected in their anti-tumor efficacy. It is expected that the new information gathered will be invaluable in further molecular design of recombinant anti-cancer cytotoxins.

描述（由申请人提供）：在绝大多数高级神经胶质瘤（HGG）患者中发现了白介素 13（IL13）的限制性受体（R）。 IL13 的 HGG 相关受体被分子鉴定为 IL13R(2，它属于肿瘤抗原家族，称为癌症/睾丸肿瘤抗原 (CTA)。CTA 为癌症的分子靶向/识别提供高度特异性。此外，基于 IL13 的细菌毒素，含有假单胞菌外毒素 A (PE) 的重组细胞毒素被证明是临床前有效的抗神经胶质瘤药物IL13 的结构-功能关系分析及其最近揭示的溶液结构表明，α-螺旋 D 似乎在 IL13 与其 HGG- 的结合中发挥着关键作用。提议产生新型高度特异性和高效的基于 IL13 的细胞毒素，其中将结合（i）有关 IL13 结构-功能关系的新知识，（ii）有关神经胶质瘤细胞生物学的新信息， (iii) 以前在临床环境中使用细胞毒素的经验。因此，这些新的细胞毒素将由基因工程形式的 IL13 和另一种细菌毒素白喉毒素 DT390 的衍生物组成。这些细胞毒素设计背后的主要思想是生产新型细胞毒素，使 IL13 的结合区域（D 螺旋）能够自由地供 IL13 的 HGG 相关受体使用，并消除细胞因子上与 IL13 相互作用的位点。它的正常生理受体在许多重要器官中表达，包括中枢神经系统。 IL13 将被设计为使 DT 毒素部分远离其与 IL13R(2 的结合位点，这是 PE 无法完全实现的。此外，IL13 α-螺旋 D 突变体，即 IL13R(2 的超级激动剂) 已被鉴定并它们将用于优化细胞毒素的设计。细胞毒素和细胞内蛋白酶激活细菌毒素（弗林蛋白酶）的受限结合位点的数量在其中发挥着重要作用。因此，将检查其表达水平的调节，并在体外和体内测试与新型IL13-DT细胞毒素功效相关的表达水平。为了证明靶标和毒素激活部分的生物表达背景下分子设计的变化如何反映在其抗肿瘤功效中，预计收集到的新信息将得到体现。在重组抗癌细胞毒素的进一步分子设计中具有不可估量的价值。