A novel peptide assay for hepcidin clinical monitoring

一种用于铁调素临床监测的新型肽测定方法

• 批准号: 10698746
• 负责人: Martyn Darby
• 金额: $ 61.38万
• 依托单位: AFFINERGY, LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698746
• 关键词: Acute; Amino Acids; Anemia; Anemia due to Chronic Disorder; Antibodies; Antisense Oligonucleotides; Bacteriophages; Binding; Biological Assay; Blood; Calibration; Carrier Proteins; Cations; Chromatography; Chronic Kidney Failure; Clinical; Clinical Trials; Competence; Complex; Data; Detection; Development; Diagnosis; Diagnostic tests; Dietary Iron; Disease; Dose; Equilibrium; Equus caballus; FDA approved; Funding; Goals; Guidelines; Hematology; Hematopoietic; Hemochromatosis; Hereditary hemochromatosis; Hormones; Immunoassay; Individual; Inherited; Iron; Iron Metabolism Disorders; Iron Overload; Iron deficiency anemia; Isotopes; Label; Laboratories; Lead; Libraries; Life; Link; Marketing; Mass Spectrum Analysis; Measures; Methods; Monitor; Patients; Peptides; Performance; Peroxidases; Phage Display; Pharmaceutical Preparations; Phase; Plasma; Preparation; Production; Protein Isoforms; Radish; Reagent; Renal carcinoma; Reporting; Reproducibility; Sampling; Serodiagnoses; Serology test; Serum; Small Business Innovation Research Grant; Small Interfering RNA; Specificity; Streptavidin; Structure; Testing; Thalassemia; Therapeutic; Validation; Viola; beta-Galactosidase; commercialization; companion diagnostics; cross reactivity; detection limit; detection method; digital; drug development; hepcidin; immunogenicity; improved; innovative technologies; instrument; instrumentation; iron absorption; iron deficiency; iron metabolism; manufacture; manufacturing scale-up; metal transporting protein 1; nanomolar; new therapeutic target; novel; novel diagnostics; novel therapeutics; overexpression; patient response; peptide hormone; personalized care; prevent; programs; protein aminoacid sequence; research clinical testing; response; small molecule; stability testing; success; time of flight mass spectrometry; tool; toxicant; uptake; Acute; Amino Acids; Anemia; Anemia due to Chronic Disorder; Antibodies; Antisense Oligonucleotides; Bacteriophages; Binding; Biological Assay; Blood; Calibration; Carrier Proteins; Cations; Chromatography; Chronic Kidney Failure; Clinical; Clinical Trials; Competence; Complex; Data; Detection; Development; Diagnosis; Diagnostic tests; Dietary Iron; Disease; Dose; Equilibrium; Equus caballus; FDA approved; Funding; Goals; Guidelines; Hematology; Hematopoietic; Hemochromatosis; Hereditary hemochromatosis; Hormones; Immunoassay; Individual; Inherited; Iron; Iron Metabolism Disorders; Iron Overload; Iron deficiency anemia; Isotopes; Label; Laboratories; Lead; Libraries; Life; Link; Marketing; Mass Spectrum Analysis; Measures; Methods; Monitor; Patients; Peptides; Performance; Peroxidases; Phage Display; Pharmaceutical Preparations; Phase; Plasma; Preparation; Production; Protein Isoforms; Radish; Reagent; Renal carcinoma; Reporting; Reproducibility; Sampling; Serodiagnoses; Serology test; Serum; Small Business Innovation Research Grant; Small Interfering RNA; Specificity; Streptavidin; Structure; Testing; Thalassemia; Therapeutic; Validation; Viola; beta-Galactosidase; commercialization; companion diagnostics; cross reactivity; detection limit; detection method; digital; drug development; hepcidin; immunogenicity; improved; innovative technologies; instrument; instrumentation; iron absorption; iron deficiency; iron metabolism; manufacture; manufacturing scale-up; metal transporting protein 1; nanomolar; new therapeutic target; novel; novel diagnostics; novel therapeutics; overexpression; patient response; peptide hormone; personalized care; prevent; programs; protein aminoacid sequence; research clinical testing; response; small molecule; stability testing; success; time of flight mass spectrometry; tool; toxicant; uptake

SUMMARY/ABSTRACT The goal of this Phase II SBIR program is to advance the commercialization of a serological diagnostic test for hepcidin, the hormone master regulator of iron metabolism that is now the target of several therapeutics in advanced stages of clinical testing. Hepcidin is produced in response to elevated systemic iron levels and subsequently blocks dietary iron absorption. The hormone triggers the ubiquitylation of the iron transporter protein ferroportin, thereby reducing both cellular uptake and efflux of iron and encouraging its systemic clearance until reaching a healthy equilibrium. Hepcidin's function is at the center of inherited iron metabolism disorders including hemochromatosis and -thalassemia, rendering it an intriguing target for new drug development efforts. Antisense oligonucleotides, siRNAs, and small molecules targeting hepcidin regulators are currently in clinical trials to modulate hepcidin expression to ameliorate both iron overload and anemia in various clinical indications. An acute need for hepcidin diagnostic tests has therefore emerged, and once available, would likely be used by clinicians as not only a companion diagnostic for these next generation therapeutics, but also monitoring tools for patients with iron dysregulation and hematopoietic disorders. Currently there is no FDA-approved diagnostic test for quantifying hepcidin in patients, largely due to the hormone's low antigenicity and the presence of multiple isoforms, many of which are inactive and therefore clinically irrelevant. Affinergy has developed a unique peptide-based sandwich assay that will enable frequent, simple, and affordable monitoring of bioactive hepcidin-25 levels in plasma. Through a previously funded Phase I program and further internal development, Affinergy has used its core competency, phage display biopanning, to identify proprietary hepcidin-binding peptides (HBPs), unique in both sequence and structure, that specifically recognize hepcidin. Further phage display biopanning has identified a phage which uniquely recognizes the HBP-hepcidin-25 complex at nanomolar concentrations. Plate-based assays using these reagents revealed a lower-limit of detection of 4 nM, consistent with the low end of normal hepcidin levels which is generally reported as 15 ng/mL or 5 nM. The sensitivity and accuracy of the assay was shown to strongly correlate with mass spectrometry-based detection of hepcidin-25. Based on these data, the Phase II program will focus on optimizing a peptide-based assay, scaling up manufacturing of these proprietary reagents, testing for the impact of contaminants (medications, common toxicants, etc.) on assay performance, and validating the assay prior to assembling a de novo 510(k) premarketing clearance application for submission to the FDA.

摘要/摘要 该II阶段SBIR计划的目标是推进血清学诊断测试的商业化 对于肝素，铁代谢的主要调节剂，现在是几种治疗剂的靶标 临床测试的高级阶段。肝素是为了响应升高的全身铁水平而产生的 随后阻止饮食中的滥用饮食。霍斯纳触发了铁运输蛋白的泛素化 蛋白质铁托蛋白，从而减少了铁的细胞摄取和外排，并鼓励其全身性 清除直到达到健康的等效物。肝素的功能是继承的铁代谢的中心 包括血色素症和丘阿无症在内的疾病，使其成为新药的有趣靶标 发展工作。反义寡核苷酸，siRNA和针对肝素调节剂的小分子 目前正在临床试验中调节肝素表达以改善铁超负荷和贫血 各种临床适应症。因此，已经出现了对肝素诊断测试的急性需求，一次 可用的，临床医生可能不仅将其用作这些下一代的同伴诊断 治疗，还为铁失调和造血疾病的患者监测工具。 目前尚无通过FDA批准的诊断测试来量化患者的肝素，这主要是由于 马酮的低抗原性和多种同工型的存在，其中许多是不活跃的，因此 临床上无关。 Affinergy开发了一种独特的基于肽的三明治测定法 通常，简单且负担得起的血浆生物活性肝素25水平的监测。通过一个 Affinergy以前资助的第一阶段计划和进一步的内部发展，已经使用了其核心能力， 噬菌体展示生物植物，以识别专有的肝素结合肽（HBP），在两个序列中都独有 和结构，特别识别肝素。进一步的噬菌体展示生物动物已经确定了噬菌体 它独特地识别纳摩尔浓度下的HBP - 甲基蛋白-25复合物。基于板的测定 使用这些试剂显示出较低的检测4 nm，与正常肝素的低端一致 通常报告为15 ng/ml或5 nm的水平。显示了测定的灵敏度和准确性 与基于质谱的肝素-25检测密切相关。基于这些数据，阶段 II计划将着重于优化基于肽的测定法，扩大这些专有的制造 试剂，测试污染物（药物，常见毒物等）对测定性能的影响， 并在组装新的510（k）之前验证该测定法 提交给FDA。