苯丙氨酸羟化酶缺乏症治疗药物初步筛选的研究

Preliminary screening of therapeutic drugs for phenylalanine hydroxylase deficiency

目的为苯丙氨酸羟化酶缺乏症患者筛选新的治疗药物。方法2019年10月至2020年10月,南京医科大学附属妇产医院医学遗传中心应用计算机针对苯丙氨酸羟化酶和药物空间结构结合力特点进行虚拟药物筛选,从美国食品药品监督管理局(FDA)药物库(包含2697种原料药)中共筛选出10种候选老药;运用真核表达系统在分子水平上检测药物对苯丙氨酸羟化酶活性的影响,并筛选药物敏感突变体。结果10种候选老药中,盐酸奈福泮、醋酸氟轻松及利司培酮分别可增加23%[t=18.21,P<0.001,与苯丙氨酸羟化酶非药处理组(即反应体系中只加入溶剂,不加药物)相比]、21%(t=3.44,P<0.05,与苯丙氨酸羟化酶非药处理组相比)、31%(t=19.57,P<0.001,与苯丙氨酸羟化酶非药处理组相比)的苯丙氨酸羟化酶酶活,其余老药对酶活性影响较弱甚至抑制。p.D101N突变体可被利司培酮激活25%(t=15.86,P<0.001,与p.D101N突变体非药处理组相比)。结论盐酸奈福泮、醋酸氟轻松及利司培酮可作为苯丙氨酸羟化酶缺乏症的潜在治疗药物,且p.D101N突变体可作为药物敏感突变位点。

Objective To screen new drugs for treatment of phenylalanine hydroxylase deficiency.Methods From October 2019 to October 2020,virtual drug screening was performed in Center of Genetic Medicine,Nanjing Maternity and Child Health Care Hospital,Women's Hospital of Nanjing Medical University computer according to the characteristics of the binding ability of phenylalanine hydroxylase to drug spatial structure.Ten candidate drugs were screened from the FDA drug library(including 2697 kinds of active pharmaceutical ingredients).A eukaryotic expression system was used to determine the effects of drugs on the activity of phenylalanine hydroxylase at the molecular level.Drug-sensitive mutants were screened.Results Among the 10 candidate drugs,neoplasm hydrochloride,fluocinonide acetate and risperidone increased 23%[t=18.21,P<0.001,vs.non-drug-treated phenylalanine hydroxylase group(i.e.,only solvent and no drug added to the reaction system)],21%(t=3.44,P<0.05,vs.non-drug-treated phenylalanine hydroxylase group),31%(t=19.57,P<0.001,vs.non-drug-treated phenylalanine hydroxylase group)of the activity of phenylalanine hydroxylase.The remaining drugs exhibited weak even inhibitory effects on the activity of phenylalanine hydroxylase.25%of p.D101N mutant could be activated by risperidone(t=15.86,P<0.001,vs.non-drug-treated p.D101N mutant group).Conclusion Neoplasm hydrochloride,fluocinonide acetate and risperidone can be used as potential therapeutic drugs for phenylalanine hydroxylase deficiency,and p.D101N mutant can be used as the drug-sensitive mutation site.