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 H₃PO₄ in H₂O/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.

Verubecestat是一种β-位淀粉样蛋白前体蛋白裂解酶1（BACE1）的抑制剂，正在临床试验中评估其治疗阿尔茨海默氏病的能力。合成路线的开发涉及非对映选择性转化，需要确定每个中间和最终活性药物成分（API）的对映体过量（ee）。经过验证的方法的分析技术包依赖于对映选择性SFC和RPLC分离，该分离使用多个3和5μm涂层的基于多糖的手性固定相（CSP）和流动相的组合。对最近开发的手性色谱柱的评估显示，使用H ₂ PO ₄在H ₂中将单个手性选择剂（Teicoplanin）结合到2.7μm核-壳颗粒上O / ACN和乙酸三乙铵：不同等度组成下的甲醇洗脱液可很好地分离所有verubecestat中间体。可以在NicoShell（另一种与2.7μm表面多孔颗粒结合的大环糖肽手性选择剂）上容易地进行verubecestat的EE测定。该方法仅使用两个手性柱和常规HPLC系统中的流动相就可以对整个verubecestat合成路线进行快速可靠的对映体分析，从而简化了技术包装的制备，方法验证和转移至制造设施。