Coiled-coil Based Drug-Free Macromolecular Therapeutics

基于卷曲线圈的无药大分子治疗

• 批准号: 8457100
• 负责人: JINDRICH H. KOPECEK
• 金额: $ 27.33万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8457100
• 关键词: Amino Acids; Animal Model; Antibodies; Antigen-Antibody Reactions; Antigens; Apoptosis; B-Cell NonHodgkins Lymphoma; B-Lymphocytes; Binding; Biological; Biological Assay; Biological Markers; Biological Process; CD20 Antigens; Cell Line; Cell surface; Cells; Charge; Cholecystokinin; Data; Diagnosis; Dose; Evaluation; Event; Exposure to; Fab Immunoglobulins; Frequencies; Goals; Immune response; Immunoglobulin Fragments; In Vitro; Incidence; Induction of Apoptosis; Lead; MS4A1 gene; Malignant Neoplasms; Mediating; Methods; Modality; Molecular Weight; Mus; Non-Hodgkin' s Lymphoma; Patients; Peptides; Pharmaceutical Preparations; Property; Protocols documentation; Roche brand of rituximab; Rodent; Structure; Surface; System; Testing; Therapeutic; Time; Toxic effect; Translating; Treatment Efficacy; United States; Validation; Vertebral column; base; biomaterial compatibility; cell type; copolymer; crosslink; design; improved; in vivo; methacrylamide; novel; novel strategies; novel therapeutics; receptor; therapeutic target; tositumomab; treatment strategy; tumor

DESCRIPTION (provided by applicant): Non-Hodgkin's lymphoma (NHL) is a prevalent cancer with an estimated 66,000 cases diagnosed in the United States in 2008, with the incidence doubling since 1980. Although treatments for NHLs greatly improved following the FDA approval of Rituxan, refractive malignancies still occur that are nonresponsive to current therapies in at least a third of all patients, indicating that improved treatment strategies are needed. One of the most reliable biomarkers and therapeutic targets for B cell NHL is CD20, which is a non- internalizing antigen that remains on the cell surface when bound to a complementary antibody. The crosslinking of CD20-bound antibodies with a secondary antibody results in apoptosis of these cells. The overall goal of this project is to generate new, drug-free macromolecular therapeutics for improved treatment of NHL. A new apoptosis induction system is proposed which is based on N-(2- hydroxypropyl)methacrylamide (HPMA) graft copolymer mediated formation of coiled-coil heterodimers at B- cell surface. The rationale of the design is the absence of low molecular weight drugs and the fact that crosslinking of CD20 at B-cell surface results in apoptosis. The system is composed of a pair of complementary coiled-coil peptides forming antiparallel heterodimers; Fab' fragment of the 1F5 anti-CD20 antibody; and HPMA copolymer. One peptide is conjugated to the Fab' fragment, the other is conjugated in multiple grafts to polyHPMA. A pair of oppositely charged pentaheptad peptides (CCE and CCK) that formed antiparallel coiled-coil heterodimers with a high degree of biorecognition was previously designed. It was hypothesized that the unique biorecognition of CCK and CCE peptides could be a basis for the design of a novel class of macromolecular therapeutics. Indeed, the exposure of CD20+ Raji B cells to Fab'-CCE resulted in the decoration of the cell surface with multiple copies of the CCE peptide via antigen-antibody fragment biorecognition. Further exposure of the CCE decorated cells to HPMA copolymer grafted with multiple copies of CCK resulted in the formation of CCE-CCK coiled-coil heterodimers on the cell surface. This second biorecognition event induced crosslinking of CD20 receptors and triggered apoptosis of Raji B cells. The system will be optimized based on the preliminary data. Shorter coiled-coil forming sequences containing L- and/or D-amino acid residues will be designed and evaluated. HPMA graft copolymer mediated coiled-coil formation at cell surface with concomitant receptor crosslinking and apoptosis induction will be assessed in vitro using five cell types and three apoptosis assays. Importantly, the efficacy and biocompatibility of drug-free macromolecular therapeutics will be evaluated in an animal model. Body distribution, impact of the dose and frequency of administration on the efficacy of treatment will be evaluated with the aim to select leading compound(s) and optimal treatment modalities. Finally, this new concept will be developed into a new therapeutic entity, drug-free macromolecular therapeutics.

描述（由申请人提供）：非霍奇金的淋巴瘤（NHL）是一种普遍的癌症，估计有66,000例在2008年在美国诊断出的病例，自1980年以来的发病率增加了一倍。尽管NHL的治疗方法大大改善了FDA在FDA的批准之后，在RITAUXAN的批准中仍然存在不利的extrive，这是当前的鉴定，这是当前的鉴定，这是当前的鉴定，这一迹象是不断提高的。需要改进的治疗策略。 B细胞NHL最可靠的生物标志物和治疗靶标之一是CD20，它是一种非内在抗原，当与互补抗体结合时，它保持在细胞表面上。与二抗CD20结合的抗体的交联导致这些细胞凋亡。该项目的总体目标是生成新的，无药物的大分子疗法，以改善NHL的治疗。提出了一种新的凋亡诱导系统，该系统基于N-（2-羟基丙烯）甲基丙烯酰胺（HPMA）接枝共聚物共聚物介导的B-细胞表面盘绕螺旋杂化二聚体的形成。设计的理由是缺乏低分子量药物，而在B细胞表面上CD20的交联导致凋亡。该系统由形成反平行异二聚体的一对互补卷曲螺旋肽组成。 1F5抗CD20抗体的Fab'片段；和HPMA共聚物。一种肽与Fab的片段偶联，另一种肽在多个移植物中与多HPMA结合。先前设计了一对相对充满电的五脑肽肽（CCE和CCK），该肽（CCE和CCK）以前设计了具有高度生物识别的反平行盘绕螺旋形二聚体。假设CCK和CCE肽的独特生物认识可能是设计新型大分子疗法的基础。实际上，CD20+ Raji B细胞暴露于Fab'-CCE导致细胞表面通过抗原抗体 - 抗体片段片段的生物认识进行了多个CCE肽的装饰。 CCE装饰的细胞进一步暴露于用多个CCK副本接枝的HPMA共聚物接触，从而导致在细胞表面形成CCE-CCK串联螺旋形的异二聚体。第二次生物识别事件诱导了CD20受体的交联，并触发了Raji B细胞的凋亡。该系统将根据初步数据进行优化。将设计和评估含有L-和/或D-氨基酸残基的较短的盘绕螺旋形成序列。 HPMA移植物共聚物介导的在细胞表面的盘绕螺旋形成，并将使用五种细胞类型和三种细胞凋亡分析在体外评估受体交联和凋亡诱导。重要的是，将在动物模型中评估无药物大分子疗法的功效和生物相容性。将评估身体分布，剂量和给药频率对治疗功效的影响，目的是选择铅化合物和最佳治疗方式。最后，这个新概念将发展成为一个新的治疗实体，无药物的大分子治疗剂。