IVD EIT: A Nonanimal Kit and Service for the Detection of Reversible Ocular Irritants

摘要 目前，Draize活兔眼测试用于区分消费品和材料， 安全免受那些危险的，可能导致可逆或不可逆的眼睛损伤。但新 法律目前正在或计划在加州、美国和欧洲实施， 欧盟禁止在眼刺激试验中使用动物。尽管如此，目前还没有单一的 非动物试验或非动物试验的组合，可以准确地分类眼刺激物。特别令人 关注所有非动物试验通常高估了最小损害性的眼部刺激物（EPA IV和 GHS未分类材料），无法确定（或以低灵敏度确定）最极端的类别 (EPA I和GHS 1不可逆/腐蚀性材料），它们的灵敏度可低至50%。 更重要的是，目前验证的替代试验都不能检测可逆刺激物（EPA II/III 和GHS 2A/B）或引起持续7至21天刺激但随后清除而不 永久性角膜损伤这一目标分类代表了正确告知 并向消费者和制造商提供有关产品安全的建议。我们最近开发并测试了一种 体外损伤深度眼刺激试验（IVD EIT）是一种检测可逆性眼刺激物的新方法， 灵敏度和特异性高。本申请的目的是进一步开发IVD EIT， 长期目标是提供一种服务和/或产品套件，用于对可逆性眼刺激物进行准确分类， 作为非动物眼刺激试验策略的一部分，满足该未满足需求。这一目标将通过 实现以下具体目标：目标1（2个月）：评估 IVD EIT通过对由EPA I-IV组成的四种化学测试集进行重复测量实验来实现 化学品来确定实现高重复性所需的角膜和阳性对照的数量。目的 2（4个月）：进行预验证试验，以评估IVD EIT对30种供试物质进行分类的能力 根据GHS和EPA分类，进一步完善预测模型。目标2将具体 确定IVD EIT的准确性、灵敏度和特异性，并确定任何应用领域 局限性（检测到的刺激物类别和化学/物理特性的范围）。如果目标2的结果 验证并证明IVD EIT的商业和监管实用性，如检测能力所定义 可逆刺激物，我们将提出第二阶段SBIR项目。 1

Abstract Currently, the Draize live rabbit eye test is used to distinguish consumer products and materials that are safe from those that are hazardous and can cause reversible or irreversible ocular damage. However, new laws are currently being, or are planned to be, implemented in California, the United States, and the European Union to ban the use of animals in ocular irritation testing. Despite this fact, there is currently no single nonanimal test or combination of nonanimal tests that can accurately classify ocular irritants. It is of particular concern that all nonanimal tests generally overpredict the least damaging class of ocular irritants (EPA IV and GHS nonclassified materials) and cannot determine (or determine with low sensitivity) the most extreme class (EPA I and GHS 1 irreversible/corrosive materials) for which they can have sensitivity rates as low as 50%. More importantly, none of the currently validated alternative tests can detect either reversible irritants (EPA II/III and GHS 2A/B) or those materials that cause irritation that lasts from 7 to 21 days but then clears without permanent corneal damage. This target classification represents a critical UNMET NEED for correctly informing and advising consumers and manufacturers about product safety. We have recently developed and tested a new method—the in vitro depth of injury eye irritation test (IVD EIT) to detect reversible ocular irritants with high sensitivity and specificity. The purpose of this application is to further develop the IVD EIT with the long- term goal of providing a service and/or product kit for the accurate classification of reversible ocular irritants to meet this UNMET NEED as part of a nonanimal ocular irritation testing strategy. This goal will be achieved by accomplishing the following specific aims: Aim 1 (2 months): Assess the repeatability and reproducibility of the IVD EIT by performing a repeat-measures experiment of a four-chemical test set composed of EPA I–IV chemicals to determine the number of corneas and positive controls needed to achieve high repeatability. Aim 2 (4 months): Perform a prevalidation test to assess the ability of the IVD EIT to categorize 30 test substances according to GHS and EPA classifications and further refine the prediction model. Aim 2 will specifically determine the accuracy, sensitivity, and specificity of the IVD EIT and identify any application domain limitations (the range of irritant categories and chemical/physical properties detected). If the results from Aim 2 validate and demonstrate the commercial and regulatory utility of the IVD EIT as defined by the ability to detect reversible irritants, we would then propose a phase II SBIR project. 1