Interactions of natural and synthetic compounds with steroidogenic enzymes and sex hormone receptor signalling

社会对可能扰乱内分泌过程、可能导致生殖问题、某些癌症和其他与（性）分化、生长和发育有关的毒性的化学品的关注日益增加。已经鉴定出多种化合物作为类固醇生成的抑制剂，包括杀虫剂、药物和天然化合物。其中大多数是体外研究，很少有体内研究。芳香酶（CYP 19）和类固醇5 α-还原酶（SRD 5A）是特别感兴趣的，因为它们是分别产生最有效的雌激素和雄激素（17 β-雌二醇和DHT）的关键类固醇生成酶，并且参与各种内分泌病理学。我们假设，天然或合成的化合物与内源性雄激素（雄烯二酮，睾酮）的结构相似，作为CYP 19和SRD 5A活性（利用雄激素作为天然底物）的抑制剂，并作为雄激素受体（AR）的拮抗剂。我的研究计划的目标是：1）确定天然或合成化学品的结构要求，使其能够干扰CYP 19和SRD 5A的催化活性或基因表达，以及AR信号传导; 2）阐明机制在基于细胞的测定中，体外酶抑制/诱导或AR抑制（激动）（涉及的信号传导途径）; 3）在生物发光小鼠模型中评价这些化合物的体内生物活性。应用纳米技术，在体外具有有效和可能有益的效果，但在体内效果不佳的化合物将被封装在纳米颗粒中，这些纳米颗粒具有改善体内递送和生物功效的特性。这项研究计划将提供一个更好的理解机制（信号通路）参与CYP 19，SRD 5A和AR调节。它将确定新的结构，可能导致开发新的CYP 19和SRD 5A抑制剂或AR拮抗剂，可用于治疗各种疾病。它将评价或改善体外显示有活性的化合物的体内有效性，提供关于将体外效应外推至体内情况的有效条件的重要信息。

There is increasing societal concern regarding chemicals that may disrupt endocrine processes, potentially resulting in reproductive problems, certain cancers and other toxicities related to (sexual) differentiation, growth and development. Various compounds have been identified as inhibitors of steroidogenesis, including pesticides, drugs and natural compounds. Most of these are in vitro studies, few in vivo studies exist. Aromatase (CYP19) and steroid 5alpha-reductase (SRD5A) are of particular interest as they are the key steroidogenic enzymes producing the most potent estrogens and androgens, 17ß-estradiol and DHT, respectively, and are involved in various endocrine pathologies. We hypothesize that natural or synthetic compounds with structures resembling the endogenous androgens (androstenedione, testosterone) act as inhibitors of CYP19 and SRD5A activity (which utilize androgens as natural substrates) and as antagonists of the androgen receptor (AR). The objectives of my research programme are to 1) identify the structural requirements of natural or synthetic chemicals enabling them to interfere with the catalytic activity or gene expression of the CYP19 and SRD5A, as well as with AR signalling; 2) elucidate mechanisms (signalling pathways involved) of enzyme inhibition/induction or AR ant(agonism) in vitro in cell-based assays; 3) evaluate the biological activity of these compounds in vivo in bioluminescent mouse models. Applying nanotechnology, compounds with potent and possibly beneficial effects in vitro, but poorly effective in vivo, will be encapsulated in nanoparticles with properties that will improve in vivo delivery and biological efficacy. This research programme will provide a better understanding of the mechanisms (signalling pathways) involved in CYP19, SRD5A and AR regulation. It will identify novel structures that may lead to the development of novel CYP19 and SRD5A inhibitors or AR antagonist which could be used to treat various diseases. It will evaluate or improve the effectiveness in vivo of compounds shown to be active in vitro providing important information on the conditions under which it is valid to extrapolate in vitro effects to an in vivo situation.