Elucidating Chemical Features that Block or Facilitate Passage across the Blood-Testis and/or Blood-Epidydimal Barriers in Mice

摘要 开发一种小分子男性避孕药因血-睾丸屏障(BTB)和血液-睾丸屏障而变得复杂。 附睾屏障(BEB)，因为许多男性避孕靶定位于 生精上皮或附睾管腔内的精子。更好地理解 使外源生物能够穿越BTB或BEB的化学特征将有助于发现和 开发新的男性避孕药。已发表的数据和我们对新型溴域睾丸(BRDT)的研究 抑制剂表明，有可能确定促进BTB或BEB通过的特性。我们会 确定促进药物在两个独立的R61中通过BTB和BEB的关键化学特征 和R33相。在R61阶段，我们将1)优化测量鼠标的协议和吞吐量 使用药物测试组的组织分布；以及2)测量100种化合物及其 代谢物和识别影响睾丸转运的分子描述符。我们将用化学方法来量化 小鼠血浆、睾丸网液(RTF)、睾丸和附睾液中的无关化合物 色质联用(LC-MS)。复合空间分布在睾丸网状部、睾丸和 将使用成像MS绘制附睾图。我们将使用化学信息学来关联结构和 实验RTF/血浆、睾丸/血浆和附睾/血浆比率的分子描述符 找出促进或阻碍睾丸堆积的特征。在R33阶段，我们计划扩大我们的 了解并直接测试改变关键推断的物理性质或分子的效果 描述符。我们将使用一系列合理设计的合成材料来验证已识别的化学特征 类比。分析化合物系列将使我们能够区分物理化学参数的趋势 以及由特定化学类型赋予的特殊性质。我们还将测试另外200部小说Small 识别影响睾丸摄取的新化学类型的分子，并测试其摄取的异常化合物 利用转运蛋白基因敲除小鼠掩盖它们的物理特性，以确定 在没有转运体的情况下发生，以改进我们的BTB和BEB的物理化学渗透性模型。 这项工作将建立适用于必须在BTB或BEB之后作用的药物的预测性摄取规则，促进 识别新的男性避孕药和治疗方法，使其在免疫系统中积累- 有特权的睾丸或附睾室将是有益的。

ABSTRACT Developing a small molecule male contraceptive is complicated by the blood-testis barrier (BTB) and blood- epididymal barrier (BEB), since many male contraceptive targets are localized to the adluminal compartment of the seminiferous epithelium or to sperm within the epididymal tubule lumen. Improved understanding of the chemical features that confer the ability for xenobiotics to cross BTB or BEB will facilitate the discovery and development of new male contraceptives. Published data and our studies of novel bromodomain testis (BRDT) inhibitors suggest that it is possible to identify properties that facilitate passage across the BTB or BEB. We will identify key chemical features that facilitate the passage of drugs across BTB and BEB in two separate R61 and R33 phases. In the R61 phase, we will 1) optimize the protocols and throughput for measuring mouse tissue distribution using a drug test set; and 2) measure tissue distribution of 100 compounds and their metabolites and identify molecular descriptors that influence delivery to the testis. We will quantify chemically unrelated compounds in mouse plasma, rete testis fluid (RTF), testis, and epididymis using liquid chromatography mass spectrometry (LC-MS). Compound spatial distribution in the rete testis, testis, and epididymis will be mapped using imaging MS. We will use chemoinformatics to correlate structures and molecular descriptors with the experimental RTF/plasma, testis/plasma, and epididymis/plasma ratios to identify features that facilitate or impede testis accumulation. In the R33 phase we plan to broaden our understanding and directly test the effects of altering the key inferred physical properties or molecular descriptors. We will 3) validate identified chemical features using a series of rationally designed synthetic analogs. Analyzing compound series will allow us to distinguish between trends in physicochemical parameters and special properties conferred by a particular chemotype. We will 4) test an additional 200 novel small molecules to identify new chemotypes that influence testis uptake and 5) test outlier compounds whose uptake belies their physical properties using transporter knock-out mice, to determine the tissue distribution that occurs without transporters so as to improve our physicochemical permeability models for the BTB and BEB. This work will establish predictive uptake rules applicable to drugs that must act behind BTB or BEB, facilitating the identification of new male contraceptives and of therapeutics for which accumulation in the immune- privileged compartments of the testis or epididymis would be beneficial.