In vivo Evaluation of Safety and Pharmacology of the Sustained Release Formulation of Dolutegravir in Pre-Conception and Early Stages of Pregnancy in Animal Models

标题：度鲁特韦缓释制剂的安全性和药理学的体内评价 怀孕和怀孕早期 摘要： 抗逆转录病毒（ARV）的可注射长效（LA）制剂代表了改善抗逆转录病毒治疗的重要替代方案。 坚持艾滋病毒/艾滋病的治疗和预防。Dolutegravir（DTG）是一种高效的ARV药物， 毒性，耐受性改善，药物间相互作用更好，副作用低，遗传屏障高， 阻力由于其优异的特性，dolutegravir被广泛用作HIV的ARV疗法的一部分。 最近，我们使用dolutegravir开发了一种超LA，可移动系统，可提供高达9 几个月，可以安全地取出以停止药物输送。尽管这种方法可能会 用于HIV治疗和预防的超LA药物递送的有效策略，长期暴露于ARV， 尤其是在怀孕期间，会引发安全问题。最近的发现加剧了这些担忧 在妊娠早期阶段对妇女进行基于DTG的治疗可能与几例 严重的神经管缺陷（NTDs）的儿童，其母亲正在接受DTG治疗。因此，至关重要的是， 使用相关体内试验系统评估长期暴露于度鲁特韦的致畸潜力 模型小鼠是一种理想的动物模型，因为它们可以快速评价药物作用，易于获得， 胚胎，以及分析药物水平，这在人类中是不可能的。我们将使用近交系小鼠品系， 对NTD（BALB/cJ、C57 BL/6 J和FVB/NJ）的不同敏感性作为准确评价 在与DTG在人体中使用最相关的条件下长期DTG暴露的相对风险： (i)单次注射长效DTG制剂后的DTG暴露，该制剂旨在改善对 人类的药物方案;（ii）每日口服DTG后长期暴露于所有当前ARV 方案是口服;（iii）暴露于DTG在孕前和怀孕期间。我们将使用急性暴露于 DTG在胚胎发育的关键阶段相当于人类妊娠第3、4、5或6周获得 深入了解怀孕期间潜在的DTG作用机制。具体而言，该分析将能够 确定并分类在人群中观察到的广泛的潜在致畸作用， 原肠胚形成的发育阶段和神经胚形成的开始，神经管闭合， 肢体发育，以及神经管闭合后的阶段，包括腭的形成。此外，本发明还提供了一种方法， 我们将评估叶酸的作用，它是NTD中最关键的因素之一。这些数据将 在评估和解释未来几年可能出现的人类出生缺陷数据方面至关重要。 年我们还将提供母体血浆、胎盘、 羊水和胚胎组织在长期每日口服DTG给药期间，在单剂量长- 在胚胎发育的关键阶段急性口服DTG剂量后。这 将使我们能够将胚胎组织中DTG的浓度与观察到的出生缺陷联系起来。评价 使用小鼠品系长期口服DTG和长效DTG制剂致畸作用 对NTD的不同敏感性代表了一种新的有价值的方法来证明安全性 DTG在怀孕前和怀孕早期阶段。

Title: In vivo evaluation of safety and pharmacology of a sustained release formulation of dolutegravir in pre- conception and early stages of pregnancy Abstract: Injectable long-acting (LA) formulations of antiretrovirals (ARVs) represent an important alternative to improve adherence to HIV/AIDS treatment and prevention. Dolutegravir (DTG) is a highly effective ARV drug with low toxicity, improved tolerability, better drug–drug interaction profile, low side-effects, and high genetic barrier to resistance. Due to its excellent properties, dolutegravir became widely used as part of ARV therapies for HIV. Recently, we used dolutegravir for development of an ultra-LA, removable system that delivers drug for up to 9 months and can be safely removed to stop drug delivery. Although this approach represents a potentially effective strategy for the ultra-LA drug delivery for HIV treatment and prevention, long-time exposure to ARV, especially during pregnancy, raises questions of safety. These concerns are exacerbated by the recent discovery that DTG-based treatment for women in early stages of pregnancy may be associated with several cases of severe neural tube defects (NTDs) in children whose mothers were being treated with DTG. It is thus vital to systematically assess the teratogenic potential of long-term exposures to dolutegravir using relevant in vivo models. Mice are an ideal animal model because they allow for rapid evaluation of drug effects, easy access to embryos, and analysis of drug levels that is not possible in humans. We will use inbred mouse strains with differential sensitivity to NTDs (BALB/cJ, C57BL/6J, and FVB/NJ) as tools for an accurate evaluation of the relative risks of long-term DTG exposure under conditions that are most relevant to the use of DTG in humans: (i) DTG exposure after a single injection of the long-acting DTG formulation designed to improve adherence to drug regimen in humans; (ii) long-term exposure to DTG after daily oral administration as all current ARV regimens are oral; (iii) exposure to DTG in preconception and during pregnancy. We will use acute exposure to DTG at critical stages of embryonic development equivalent to human pregnancy at weeks 3, 4, 5, or 6 to gain insight into the mechanism of potential DTG action during pregnancy. Specifically, this analysis will be able to identify and classify a wide spectrum of potential teratogenic effects observed in human populations in developmental stages of gastrulation and the beginning of neurulation, neural tube closure, the beginning of limb development, and stages following neural tube closure, including palate formation. In addition, we will evaluate the role of the folic acid, one of the most critical factors involved in NTDs. These data will be critical in evaluating and interpreting the human birth defects data that will likely emerge over the next several years. We will also provide a comprehensive analysis of DTG concentration in maternal plasma, placenta, amniotic fluid and embryonic tissues during chronic daily oral DTG administration, after a single dose of a long- acting formulation of DTG and after an acute oral dose of DTG at critical stages of embryonic development. This will allow us to correlate concentration of DTG in embryonic tissues with observed birth defects. Evaluation of teratogenic effect of long-term oral administration of DTG and a long-acting DTG formulation using mouse strains with differential sensitivities to NTD represents a novel and valuable approach to demonstrate the safety profile of DTG in pre-conception and during early stages of pregnancy.