DDT-BMQ-000109, Qualification of biomarkers for in vitro developmental toxicity screening in a human system

DDT-BMQ-000109，人体系统体外发育毒性筛选生物标志物的资格

• 批准号: 10836889
• 负责人: Jessica A Palmer
• 金额: $ 24.99万
• 依托单位: STEMINA BIOMARKER DISCOVERY, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10836889
• 关键词: DDT; BMQ; 000109; Qualification; biomarkers

Project Summary/Abstract Stemina Biomarker Discovery, Inc. (Stemina) has developed an in vitro human pluripotent stem cell-based assay, devTOX quickPredict (devTOXqP), that uses a biomarker ratio of ornithine to cystine to predict if a compound has the potential to cause developmental toxicity over a wide range of exposures. The goal of this U01 cooperative agreement project is to qualify the devTOXqP assay’s metabolite ratio of ornithine to cystine (o/c ratio) through the Center for Drug Evaluation and Research (CDER) Biomarker Qualification Program (BQP). Stemina has an accepted Letter of Intent (LOI) to qualify the devTOXqP o/c ratio as a safety biomarker for detecting human developmental toxicity potential in vitro using human induced pluripotent stem (iPS) cells at the nonclinical stage of drug development for small molecule drugs as part of a weight-of-evidence assessment as described in the ICH S5(R3) guideline recently issued by the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). In Aim 1, we plan to conduct a fit-for-purpose analytical method validation study for the ultra-performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) method used for measuring the o/c ratio to assess the precision, sensitivity, specificity, dilution linearity, reinjection reproducibility, extraction recovery, and sample carryover of the UPLC-HRMS analytical method. We also plan to measure test article concentrations and characterize test compound stability in the cell culture medium for a subset of the test compounds used in determining the relevance of the biomarker (Aim 5). A prospective qualification study will be conducted to: (1) determine the within-laboratory repeatability and reproducibility of the o/c ratio, (2) evaluate the reliability of the o/c ratio in multiple human iPS cell lines based on prediction concordance and the correlation of response to compound exposure, and (3) assess the relevance of the o/c ratio for predicting developmental toxicity potential across a broad range of pharmaceutical compounds (Aims 2, 3 and 6). Finally, we plan to establish a transfer plan with Stemina’s distribution partner to assess between-laboratory transferability and reliability of the of the devTOXqP standard operating procedures for the inter-laboratory portion of the Qualification Plan (Aim 4). Stemina’s partner will provide financial support for its part of the Qualification Plan. The studies proposed in this grant, and the in-kind support of Stemina’s distribution partner, will provide the necessary data for completing the studies described in the Qualification Plan (once approved) and submitting the Full Qualification Package. While in vitro assays such as devTOXqP will never entirely replace animals when used alone, these assays can reduce the number of compounds tested in animals. The assay may also replace the need for a second species in certain categories of compounds when used as part of a weight of evidence approach as described in the S5 (R3) guideline. Qualification of the devTOXqP assay for developmental toxicity assessment will reduce animal use and provide a human-based assessment of developmental toxicity, ultimately leading to safer drugs and fewer birth defects from chemical exposure in utero.

项目概要/摘要 Stemina Biomarker Discovery, Inc. (Stemina) 开发了一种基于体外人类多能干细胞的 devTOX QuickPredict (devTOXqP) 检测，使用鸟氨酸与胱氨酸的生物标志物比率来预测是否 该化合物在广泛的接触范围内有可能引起发育毒性。此举的目标 U01合作协议项目旨在验证devTOXqP检测的鸟氨酸与胱氨酸代谢物比率（o/c 比）通过药物评价和研究中心（CDER）生物标志物资格计划（BQP）。 Stemina 拥有一份公认的意向书 (LOI)，将 devTOXqP o/c 比率限定为安全生物标志物 使用人类诱导多能干 (iPS) 细胞在体外检测人类发育毒性潜力 小分子药物药物开发的非临床阶段，作为证据权重评估的一部分 国际技术协调委员会最近发布的 ICH S5(R3) 指南中描述 人用药品 (ICH) 的要求。在目标 1 中，我们计划进行适合目的的分析 超高效液相色谱-高分辨质谱方法验证研究 (UPLC-HRMS)方法用于测量o/c比以评估精密度、灵敏度、特异性、稀释度 UPLC-HRMS 分析的线性度、回样重现性、萃取回收率和样品残留 方法。我们还计划测量测试物品的浓度并表征测试化合物在细胞中的稳定性 用于确定生物标志物相关性的测试化合物子集的培养基（目标 5）。 将进行前瞻性资格研究：(1) 确定实验室内的可重复性和 o/c 比率的重现​​性，(2) 基于以下方法评估多种人类 iPS 细胞系中 o/c 比率的可靠性 预测一致性和化合物暴露反应的相关性，以及（3）评估相关性 用于预测多种药物化合物潜在发育毒性的 O/C 比 （目标 2、3 和 6）。最后，我们计划与 Stemina 的分销合作伙伴建立一个转移计划来评估 devTOXqP 标准操作程序的实验室间可转移性和可靠性 资格计划的实验室间部分（目标 4）。 Stemina的合作伙伴将为其提供财务支持 资格计划的一部分。这笔赠款中提出的研究以及 Stemina 分配的实物支持 合作伙伴，将提供完成资格计划中描述的研究所需的数据（一旦 批准）并提交完整的资格证书。虽然 devTOXqP 等体外检测永远不会 当单独使用时完全替代动物，这些测定可以减少在动物中测试的化合物的数量。 当用作某些化合物类别时，该测定还可以取代对第二种物质的需要。 S5 (R3) 指南中描述的证据权重方法的一部分。 devTOXqP 检测的资格 用于发育毒性评估将减少动物使用并提供基于人类的评估 发育毒性，最终导致药物更安全，并减少子宫内化学暴露造成的出生缺陷。