Uniquely constrained peptides for modulating TNF receptor activity

用于调节 TNF 受体活性的独特限制肽

基本信息

批准号：
10387498
负责人：
Ashley M. Kretsch
金额：
$ 6.76万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-16 至 2023-02-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10387498
关键词：
Affinity Agonist Anabolism Anti-Bacterial Agents Antibodies Antigens Area Autoimmune Autoimmune Diseases Binding Binding Sites Biological Biological Assay Biological Products Biology CD19 gene Cell Surface Receptors Cells Characteristics Chemicals Chemistry Clinical Communication Complement Consumption Cyclic Peptides Data Development Disease Educational process of instructing Endothelin Enzymes Exhibits Fellowship Generations Genetic Genome Genomics Glucagon Receptor Goals Health Human Humira Illinois Immune Targeting Immunologic Receptors In Vitro Individual Institutes Lasso Learning Libraries Ligands Mentors Mentorship Messenger RNA Methods Modality Modification Monoclonal Antibodies Mutagenesis OX40 Oral Peptide Conformation Peptide Hydrolases Peptide Library Peptides Pharmaceutical Preparations Pharmacologic Substance Positioning Attribute Post-Translational Protein Processing Property Research Personnel Research Training Resistance Resources Ribosomes Safety Scientist Shapes Signal Transduction Site Solvents Specificity Structure Structure-Activity Relationship Students Surface T-Cell Activation TNF gene TNFSF4 gene Therapeutic Therapeutic Monoclonal Antibodies Time Toxic effect Training Tumor Necrosis Factor Receptor Universities Variant Work Work Ethic adalimumab analog antagonist anti-PD-1 anti-cancer base cancer therapy career design effective therapy graduate student high throughput screening improved member next generation novel pembrolizumab peptide I rational design receptor binding screening small molecule small molecule therapeutics success targeted agent three dimensional structure tumor necrosis factor ligand superfamily member 4 undergraduate student unnatural amino acids

项目摘要

PROJECT SUMMARY There is a steady rise in the clinical use of biologics, particularly monoclonal antibodies (mAbs). We present lasso peptides as a potential drug modality within the underleveraged region of chemical space between mAbs and small molecules. Lasso peptides represent a class of ribosomally-synthesized and post-translationally modified peptide (RiPP). Biosynthesis of lasso peptides involves a genetically encoded precursor peptide and maturation enzymes that catalyze a macrolactam ring and configure the peptide into a unique lariat knot. Lasso peptides recapitulate the desired properties of synthetic cyclic peptide but with key advantages: (i) lasso peptides are uniquely constrained, highly stable globular structures with the correct shape to engage peptide-binding targets, (ii) lasso peptides are entirely genetically encoded, straightforward generation of large and sequence- diverse libraries, (iii) enzymatic tolerance supports modification of the entire solvent-exposed surface, non- natural amino acids, and post-translational modifications, and (iv) the C-terminus provides a facile conjugation site for peptide display, and high-throughput screening. Due to their unique topology, lasso peptides cannot be chemical synthesized, thus we rely on the biosynthetic enzymes to thread the peptide. This proposal will design a biosynthesis-informed library of lasso peptides and as a proof-of-concept, use this library to select for lasso peptides that bind OX40. OX40 is an immunostimulatory member of the tumor necrosis factor receptors (TNFR), studies have found that the native OX40 ligand, OX40L, and agonist antibodies can activate OX40 to stimulate the proliferation and activation of T cells. There is a compelling need to validate immune targets so that therapeutics can be developed, complementing the success of anti-PD1 (ie Keytruda) and anti- TNFα (ie Humira) biologics. Our group has developed a high-throughput method with cell-free biosynthesis (CFB) to produce lasso peptide libraries. In Aim I we will combine CFB with mRNA display to identify library members that are successfully cyclized. Aim II will select for lasso peptides that bind OX40. We will assess structure-activity relationships by validating the three-dimensional structure of the OX40-binding variants and conduct binding affinity studies and in vitro signaling assays to evaluate their activity. Motifs related to OX40 binding and activity will be identified through structural and mutagenesis data. Improved understanding of receptor binding-activity relationship will significantly inform pharmaceutical efforts. This fellowship will provide the necessary training to prepare me for an academic career in mentoring researchers and teaching students. My goal is to develop my abilities to become a more comprehensive scientist through improving communication in both oral and written form, a strong collaborative work ethic, and mentorship of undergraduate and graduate students. Working with my sponsor Douglas Mitchell, an expert in RiPPs biosynthesis, and the resources available at the University of Illinois at Urbana-Champaign, the Department of Chemistry, and the Institute for Genomic Biology, will provide an unrivaled chance to succeed in aims of the research and the training plan.

项目摘要生物制剂的临床使用，尤其是单克隆抗体（mAb）的临床使用稳步上升。我们在场拉索肽是mab之间化学空间不足区域内的潜在药物形态和小分子。拉索·佩蒂斯（Lasso Petides 修饰肽（RIPP）。拉索辣椒的生物合成涉及遗传编码的前体胡椒和成熟的酶，可催化大花酰胺环并将肽构成独特的幼虫结。套索肽概括了合成循环胡椒的所需特性，但具有关键优势：（i）拉索·佩蒂德斯（Lasso Petides）是独特的约束，高度稳定的全球结构，具有正确的形状以吸引肽结合靶标，（ii）套索肽完全是遗传编码的，直接生成大的序列 - （III）酶耐受支持整个溶液暴露的表面，非 - 天然氨基酸和翻译后修饰，（iv）C-末端提供了轻松的共轭用于辣椒展示的站点和高通量筛选。由于它们独特化学合成，因此我们依靠生物合成酶将胡椒粉丝纹。该提议将设计套索辣椒的生物合成图书馆，作为概念证明，请使用此库选择结合OX40的拉索肽。 OX40是肿瘤坏死因子的免疫刺激剂成员受体（TNFR），研究发现，天然OX40配体，OX40L和激动剂抗体可以激活 OX40刺激T细胞的增殖和激活。有迫切需要验证免疫靶标可以开发治疗，完成抗PD1（IE KEYTRUDA）和抗 - 的成功 TNFα（IE Humira）生物制剂。我们的小组开发了一种具有无细胞生物合成的高通量方法（CFB）生产套索肽库。在目标我我将结合CFB与mRNA显示以识别库成功自行车的成员。 AIM II将选择结合OX40的拉索肽。我们将评估通过验证OX40结合变体的三维结构，结构活性关系进行结合亲和力研究和体外信号传导测定以评估其活性。与OX40相关的主题结构和诱变数据将确定结合和活性。改善了对接收器结合活性关系将大大为药物工作提供依据。这个奖学金将提供必要的培训，使我为心理研究人员和教学学生的学术生涯做好准备。我的目标是通过改善沟通来发展自己成为更全面的科学家的能力以口头和书面形式，强大的协作职业道德以及本科和毕业生的指导学生。与我的赞助商道格拉斯·米切尔（Douglas Mitchell）合作，Ripps Biosynesis的专家和资源可在伊利诺伊大学的Urbana-Champaign大学，化学系和研究所获得基因组生物学，将为研究和培训计划的目标提供无与伦比的机会。