代谢异常在各种病理状况中发挥着关键作用，因此需要对特定代谢物进行量化以进行诊断。虽然质谱法仍然是代谢物测量的主要方法，但其有限的通量强调了对能够快速检测的生物传感器的需求。此前，我们报道了具有12个羧酸基团的柱[6]芳烃（P6AC）与1-甲基烟酰胺（1-MNA）形成主客体复合物，1-甲基烟酰胺（1-MNA）是由烟酰胺N-甲基转移酶（NNMT）在体内产生的。 P6AC 充当生物传感器，通过测量 1-MNA 结合时光诱导电子转移引起的荧光猝灭。然而，P6AC 的低灵敏度使其无法检测未纯化的生物样品中的 1-MNA。在这项研究中，我们发现具有 12 个磺酸基的 P6A (P6AS) 是 1-MNA 相互作用的特异且有效的超分子宿主，即使在生物样品中也是如此。 P6AS在水中的1-MNA结合亲和力为(5.68±1.02)×106 M-1，比P6AC高约700倍。此外，P6AS的1-MNA检测限为2.84×10-7 M，明显低于P6AC的检测限。直接将 P6AS 添加到培养基中足以量化癌细胞产生的 1-MNA。此外，该传感器甚至能够特异性检测未纯化的人尿液中的 1-MNA。因此，P6AS 能够快速、高通量地定量 1-MNA，我们的策略的进一步改进将有助于建立 NNMT 抑制剂的高通量筛选、肝脏疾病的诊断以及人类癌细胞体内成像。

Metabolic abnormalities play a pivotal role in various pathological conditions, necessitating the quantification of specific metabolites for diagnosis. While mass spectrometry remains the primary method for metabolite measurement, its limited throughput underscores the need for biosensors capable of rapid detection. Previously, we reported that pillar[6]arene with 12 carboxylate groups (P6AC) forms host-guest complexes with 1-methylnicotinamide (1-MNA), which is produced in vivo by nicotinamide N-methyltransferase (NNMT). P6AC acts as a biosensor by measuring the fluorescence quenching caused by photoinduced electron transfer upon 1-MNA binding. However, the low sensitivity of P6AC makes it impractical for detecting 1-MNA in unpurified biological samples. In this study, we found that P6A with 12 sulfonate groups (P6AS) is a specific and potent supramolecular host for 1-MNA interactions even in biological samples. The 1-MNA binding affinity of P6AS in water was found to be (5.68 ± 1.02) × 106 M-1, which is approximately 700-fold higher than that of P6AC. Moreover, the 1-MNA detection limit of P6AS was determined to be 2.84 × 10-7 M, which is substantially lower than that of P6AC. Direct addition of P6AS to culture medium was sufficient to quantify 1-MNA produced by cancer cells. Furthermore, this sensor was able to specifically detect 1-MNA even in unpurified human urine. P6AS therefore enables rapid and high-throughput quantification of 1-MNA, and further improvement of our strategy will contribute to the establishment of high-throughput screening of NNMT inhibitors, diagnosis of liver diseases, and imaging of human cancer cells in vivo.