Department of Biomedicine, University of Bergen, 5020 Bergen, Norway.
Norwegian Porphyria Centre (NAPOS), Laboratory for Clinical Biochemistry, Haukeland University Hospital, 5021 Bergen, Norway.
Mol Ther. 2020 Feb 5;28(2):677-689. doi: 10.1016/j.ymthe.2019.11.010. Epub 2019 Dec 4.
Mutations in hydroxymethylbilane synthase (HMBS) cause acute intermittent porphyria (AIP), an autosomal dominant disease where typically only one HMBS allele is mutated. In AIP, the accumulation of porphyrin precursors triggers life-threatening neurovisceral attacks and at long-term, entails an increased risk of hepatocellular carcinoma, kidney failure, and hypertension. Today, the only cure is liver transplantation, and a need for effective mechanism-based therapies, such as pharmacological chaperones, is prevailing. These are small molecules that specifically stabilize a target protein. They may be developed into an oral treatment, which could work curatively during acute attacks, but also prophylactically in asymptomatic HMBS mutant carriers. With the use of a 10,000 compound library, we identified four binders that further increased the initially very high thermal stability of wild-type HMBS and protected the enzyme from trypsin digestion. The best hit and a selected analog increased steady-state levels and total HMBS activity in human hepatoma cells overexpressing HMBS, and in an Hmbs-deficient mouse model with a low-expressed wild-type-like allele, compared to untreated controls. Moreover, the concentration of porphyrin precursors decreased in liver of mice treated with the best hit. Our findings demonstrate the great potential of these hits for the development of a pharmacological chaperone-based corrective treatment of AIP by enhancing wild-type HMBS function independently of the patients' specific mutation.
羟甲基胆素合酶 (HMBS) 突变导致急性间歇性卟啉症 (AIP),这是一种常染色体显性疾病,通常只有一个 HMBS 等位基因发生突变。在 AIP 中,卟啉前体的积累会引发危及生命的神经内脏发作,长期来看,会增加肝细胞癌、肾衰竭和高血压的风险。目前,唯一的治疗方法是肝移植,因此迫切需要有效的基于机制的治疗方法,如药理学伴侣。这些是专门稳定靶蛋白的小分子。它们可以开发成口服治疗药物,在急性发作时可以进行治疗性治疗,在无症状 HMBS 突变携带者中也可以进行预防性治疗。我们使用了一个包含 10000 种化合物的文库,鉴定出了四个结合物,它们进一步提高了野生型 HMBS 最初非常高的热稳定性,并保护了酶免受胰蛋白酶消化。最好的命中结果和一个选定的类似物增加了人肝癌细胞中 HMBS 过表达和低表达野生型样等位基因的 Hmbs 缺陷型小鼠模型中的稳态水平和总 HMBS 活性,与未治疗的对照组相比。此外,与未治疗的对照组相比,用最佳命中结果处理的小鼠肝脏中卟啉前体的浓度降低了。我们的研究结果表明,这些命中结果具有很大的潜力,可以通过增强野生型 HMBS 功能来开发基于药理学伴侣的 AIP 矫正治疗,而无需考虑患者的特定突变。