Ďuriš Andrej, Daïch Adam, Santos Cécile, Fleury Laurence, Ausseil Frédéric, Rodriguez Frédéric, Ballereau Stéphanie, Génisson Yves, Berkeš Dušan
Department of Organic Chemistry, Slovak University of Technology, Radlinského 9, 81237, Bratislava, Slovak Republic), Fax.
Normandie Univ, UNIHAVRE, CNRS, URCOM, 76600, Le Havre (France), CNRS INC3M, FR 3038, EA 3221, UFR des Sciences et Techniques, 25 rue Philippe Lebon, B.P. 1123, 76063, Le Havre Cedex, France), Fax.
Chemistry. 2016 May 4;22(19):6676-86. doi: 10.1002/chem.201505121. Epub 2016 Mar 31.
A series of 12 analogues of the Cer transfer protein (CERT) antagonist HPA-12 with long aliphatic chains were prepared as their (1R,3S)-syn and (1R,3R)-anti stereoisomers from pivotal chiral oxoamino acids. The enantioselective access to these intermediates as well as their ensuing transformation relied on a practical crystallization-induced asymmetric transformation (CIAT) process. Sonogashira coupling followed by triple bond reduction and thiophene ring hydrodesulfurization (HDS) into the corresponding alkane moieties was then implemented to complete the synthetic routes delivering the targeted HPA-12 analogues in concise 4- to 6-step reaction sequences. Ten compounds were evaluated regarding their ability to bind to the CERT START domain by using the recently developed time-resolved FRET-based homogeneous (HTR-FRET) binding assay. The introduction of a lipophilic appendage on the phenyl moiety led to an overall 10- to 1000-fold enhancement of the protein binding, with the highest effect being observed for a n-hexyl residue in the meta position. The importance of the phenyl ring for the activity was indicated by the reduced potency of the 3-deoxyphytoceramide aliphatic analogues. The 1,3-syn stereoisomers were systematically more potent than their 1,3-anti analogues. In silico studies were used to rationalized these trends, leading to a model of protein recognition coherent with the stronger binding of (1R,3S)-syn HPAs.
从关键的手性氧代氨基酸出发，制备了一系列具有长脂肪链的神经酰胺转运蛋白（CERT）拮抗剂HPA - 12的12种类似物，包括其（1R,3S）-顺式和（1R,3R）-反式立体异构体。对这些中间体的对映选择性合成及其后续转化依赖于一种实用的结晶诱导不对称转化（CIAT）过程。随后实施了Sonogashira偶联反应，接着进行三键还原以及噻吩环加氢脱硫（HDS）反应生成相应的烷烃部分，以简洁的4至6步反应序列完成合成路线，得到目标HPA - 12类似物。使用最近开发的基于时间分辨荧光共振能量转移（FRET）的均相（HTR - FRET）结合测定法，评估了10种化合物与CERT START结构域结合的能力。在苯基部分引入亲脂性取代基导致蛋白质结合能力总体提高了10至1000倍，间位的正己基残基效果最为显著。3 - 脱氧植物神经酰胺脂肪族类似物活性的降低表明了苯环对活性的重要性。1,3 - 顺式立体异构体在系统上比其1,3 - 反式类似物更具活性。通过计算机模拟研究来合理化这些趋势，从而得到一个与（1R,3S）-顺式HPA更强结合相一致的蛋白质识别模型。
