Barhate Chandan L, Lopez Diego A, Makarov Alexey A, Bu Xiaodong, Morris William J, Lekhal Azzeddine, Hartman Robert, Armstrong Daniel W, Regalado Erik L
Department of Chemistry, University of Texas at Arlington, Arlington, TX 76019, USA.
Process Research and Development, MRL, Merck & Co., Inc., Rahway, NJ 07065, USA.
J Chromatogr A. 2018 Mar 2;1539:87-92. doi: 10.1016/j.chroma.2018.01.042. Epub 2018 Jan 31.
Verubecestat is an inhibitor of β-site amyloid precursor protein cleaving enzyme 1 (BACE1) being evaluated in clinical trials for the treatment of Alzheimer's disease. Synthetic route development involves diastereoselective transformations with a need for enantiomeric excess (ee) determination of each intermediate and final active pharmaceutical ingredient (API). The analytical technical package of validated methods relies on enantioselective SFC and RPLC separations using multiple 3 and 5 μm coated polysaccharide-based chiral stationary phases (CSPs) and mobile phases combinations. Evaluation of recently developed chiral columns revealed a single chiral selector (Teicoplanin) bonded to 2.7 μm core-shell particles using HPO in HO/ACN and triethylammonium acetate: methanol based eluents at different isocratic compositions allowed good enatioseparation of all verubecestat intermediates. EE determination of verubecestat is easily performed on NicoShell, another macrocyclic glycopeptide chiral selector bonded to 2.7 μm superficially porous particles. This approach enables fast and reliable enantiopurity analysis of the entire verubecestat synthetic route using only two chiral columns and mobile phases on a conventional HPLC system, simplifying technical package preparation, method validation and transfer to manufacturing facilities.
维鲁贝西他是一种β-位点淀粉样前体蛋白裂解酶1（BACE1）抑制剂，正在进行治疗阿尔茨海默病的临床试验评估。合成路线开发涉及非对映选择性转化，需要对每个中间体和最终活性药物成分（API）进行对映体过量（ee）测定。经过验证的方法的分析技术包依赖于使用多种3μm和5μm涂覆的多糖基手性固定相（CSP）和流动相组合的对映选择性超临界流体色谱（SFC）和反相液相色谱（RPLC）分离。对最近开发的手性柱的评估表明，使用万古霉素作为单一手性选择剂，将其键合到2.7μm核壳颗粒上，在不同等度组成的水/乙腈和乙酸三乙铵：甲醇基洗脱剂中使用HPO，能够对所有维鲁贝西他中间体实现良好的对映体分离。在NicoShell（另一种键合到2.7μm表面多孔颗粒上的大环糖肽手性选择剂）上可以轻松进行维鲁贝西他的ee测定。这种方法仅使用两根手性柱和流动相，就能在传统高效液相色谱系统上对整个维鲁贝西他合成路线进行快速可靠的对映体纯度分析，简化了技术包的制备、方法验证以及向生产设施的转移。
