Lumacaftor/ivacaftor therapy is associated with reduced hepatic steatosis in cystic fibrosis patients

BACKGROUND Hepatic steatosis is a common form of cystic fibrosis associated liver disease(CFLD)seen in an estimated 15%-60%of patients with cystic fibrosis(CF).The pathophysiology and health implications of hepatic steatosis in cystic fibrosis remain largely unknown.In the general population,hepatic steatosis is strongly associated with insulin resistance and type 2 diabetes.Cystic fibrosis related diabetes(CFRD)impacts 40%-50%of CF adults and is characterized by both insulin insufficiency and insulin resistance.We hypothesized that patients with CFRD would have higher levels of hepatic steatosis than cystic fibrosis patients without diabetes.AIM To determine whether CFRD is associated with hepatic steatosis and to explore the impact of lumacaftor/ivacaftor therapy on hepatic steatosis in CF.METHODS Thirty patients with CF were recruited from a tertiary care medical center for this cross-sectional study.Only pancreatic insufficient patients with CFRD or normal glucose tolerance(NGT)were included.Patients with established CFLD,end stage lung disease,or persistently elevated liver enzymes were excluded.Mean magnetic resonance imaging(MRI)proton density fat fraction(PDFF)was obtained for all participants.Clinical characteristics[age,sex,body mass index,percent predicted forced expiratory volume at 1 s(FEV1),lumacaftor/ivacaftor use]and blood chemistries were assessed for possible association with hepatic steatosis.Hepatic steatosis was defined as a mean MRI PDFF>5%.Patients were grouped by diabetes status(CFRD,NGT)and cystic fibrosis transmembrane conductance regulator(CFTR)modulator use(lumacaftor/ivacaftor,no lumacaftor/ivacaftor)to determine between group differences.Continuous variables were analyzed with a Wilcoxon rank sum test and discrete variables with a Chi square test or Fisher’s exact test.RESULTS Twenty subjects were included in the final analysis.The median age was 22.3 years(11.3-39.0)and median FEV1 was 77%(33%-105%).Twelve subjects had CFRD and 8 had NGT.Nine subjects were receiving lumacaftor/ivacaftor.The median PDFF was 3.0%(0.0%-21.0%).Six subjects(30%)had hepatic steatosis defined as PDFF>5%.Hepatic fat fraction was significantly lower in patients receiving lumacaftor/ivacaftor(median,range)(2.0%,0.0%-6.4%)than in patients not receiving lumacaftor/ivacaftor(4.1%,2.7-21.0%),P=0.002.Though patients with CFRD had lower PDFF(2.2%,0.0%-14.5%)than patients with NGT(4.9%,2.4-21.0%)this did not reach statistical significance,P=0.06.No other clinical characteristic was strongly associated with hepatic steatosis.CONCLUSION Use of the CFTR modulator lumacaftor/ivacaftor was associated with significantly lower hepatic steatosis.No association between CFRD and hepatic steatosis was found in this cohort.

囊性纤维化治疗新药Ivacaftor的合成

Ivacaftor是被FDA批准用于由G551D基因突变引起的囊性纤维化的治疗药物。以2,4-二叔丁基苯酚为原料,经酯化保护、硝化、水解脱保护、硝基还原得到关键中间体2,4-二叔丁基-5-氨基苯酚;苯胺和乙氧亚甲基丙二酸二乙酯经Gould-Jacobs反应得到4-氧代-1,4-二氢喹啉-3-羧酸。2,4-二叔丁基-5-氨基苯酚与4-氧代-1,4-二氢喹啉-3-羧酸缩合得到Ivacaftor,总收率约13%(以苯胺计)。

A High Sensitive LC-MS/MS Method for the Simultaneous Determination of Potential Genotoxic Impurities Carboxy Phenyl Boronic Acid and Methyl Phenyl Boronic Acid in Lumacaftor

A simple, rapid, and highly sensitive LC-MS/MS method has been developed for the simultaneous and trace level quantification of underivatized boronic acids in lumacaftor active pharmaceutical ingredient. Chromatographic separation of boronic acids and lumacaftor achieved using Agilent Poroshell HPH C18 150 × 4.6 mm 2.7 μ column with 0.1% ammonia in water as mobile phase A and 100% acetonitrile as mobile phase B at a flow rate of 0.25 ml/min. Gradient elution was used with a total method run time of 14 minutes. Boronic acids were successfully ionized and quantified without derivatization using electrospray ionization in negative mode using tandem quadrupole mass spectrometry in multiple reactions monitoring mode. Method validation was performed as per ICH guidelines with good linearity over the concentration range of 0.05 ppm to 5 ppm of Lumacaftor test concentration for both the boronic acids with a correlation coefficient of >0.99. Recoveries were found good at different concentration levels and within the range of 80% - 120%. The developed method can be successfully used for the routine quantification of boronic acids at a concentration level of 20 ng/ml (1 ppm with respect to 20 mg/ml lumacaftor).

An LC-MS/MS method for simultaneous analysis of the cystic fibrosis therapeutic drugs colistin,ivacaftor and ciprofloxacin

Inhaled antibiotics such as colistin and ciprofloxacin are increasingly used to treat bacterial lung infections in cystic fibrosis patients.In this study,we established and validated a new HPLC-MS/MS method that could simultaneously detect drug concentrations of ciprofloxacin,colistin and ivacaftor in rat plasma,human epithelial cell lysate,cell culture medium,and drug transport media.An aliquot of200 μL drug-containing rat plasma or cell culture medium was treated with 600 μL of extraction solution(acetonitrile containing 0.1% formic acid and 0.2% trifluoroacetic acid(TFA)).The addition of 0.2% TFA helped to break the drug-protein bonds.Moreover,the addition of 0.1% formic acid to the transport medium and cell lysate samples could significantly improve the response and reproducibility.After vortexing and centrifuging,the sample components were analyzed by HPLC-MS/MS.The multiple reaction monitoring mode was used to detect the following transitions:585.5-101.1(colistin A),578.5-101.1(colistin B),393.2-337.2(ivacaftor),332.2-314.2(ciprofloxacin),602.3-101.1(polymyxin B1 as internal standard(IS)) and 595.4-101.1(polymyxin B2 as IS).The running time of a single sample was only 6 min,making this a time-efficient method.Linear correlations were found for colistin A at 0.029-5.82 μg/m L,colistin B at 0.016-3.14 μg/m L,ivacaftor at 0.05-10.0 μg/m L,and ciprofloxacin at 0.043-8.58 μg/m L.Accuracy,precision,and stability of the method were within the acceptable range.This method would be highly useful for research on cytotoxicity,animal pharmacokinetics,and in vitro drug delivery.

Ivacaftor关键中间体2,4-二叔丁基-5-氨基苯酚的合成工艺研究

以2,4-二叔丁基苯酚为起始原料,经亚硝化、还原两步反应制备目标产品2,4-二叔丁基-5-氨基苯酚,其结构经1H NMR和MS确证,总收率为75. 3%,粒径主要分布于20～40μm。通过单因素试验获得了较优的工艺条件,亚硝化反应:n(2,4-二叔丁基苯酚)∶n(亚硝酸钠)=1. 0∶1. 6,反应时间2～3 h;还原反应:n(2,4-二叔丁基-5-亚硝基苯酚)∶n(硫代硫酸钠)=1. 0∶2. 6,反应温度为50～60℃。该路线操作简单、反应条件温和,易于放大生产,具有较好的应用价值。

依伐卡托中间体的合成工艺改进

改进了依伐卡托(Ivacaftor)中间体2,4-二叔丁基-5-硝基苯基乙基碳酸酯的合成工艺。原料2,4-二叔丁基苯酚经过氯甲酸乙酯保护酚羟基、硝化得到中间体和2,4-二叔丁基-6-硝基苯基乙基碳酸酯的混合物,不经柱层析纯化直接利用石油醚重结晶,以56.1%的总收率制得高纯度的中间体,经HPLC测定中间体的纯度为95.4%。改进后的工艺操作简单、成本低廉、易于工业化放大,为依伐卡托中间体的合成提供了一种新的、高效的制备方法。

治疗囊性纤维化跨膜传导调节因子基因突变介导疾病的复方新药——Symdeko~

囊性纤维化是遗传性外分泌腺疾病,其跨膜传导调节因子(CFTR)是细胞膜表面一种依赖氯离子磷酸化的通道蛋白。CFTR基因突变导致蛋白产物功能破坏或丧失,所介导的疾病危及人体多个系统,如呼吸系统的慢性梗阻性肺部病变,消化系统的胰腺外分泌功能不良致使胰腺炎发作和汗液电解质异常升高等。治疗囊性纤维化跨膜传导调节因子介导疾病的复方新药Symdeko~由美国Vertex生物技术制药公司研制,含tezacaftor 100 mg和ivacaftor 150 mg的复方片剂及ivacafto 150 mg单一片剂的共包装。2018年2月12日,美国食品药品管理局(FDA)授予该药突破性治疗药物地位的认定,同日批准上市。该药同时向欧洲药品管理局(EMA)提出新药上市申请(NDA),目前尚在审理中。该文对Symdeko~非临床和临床药理毒理学、临床研究、不良反应、适应证、剂量与用法、用药注意事项及知识产权状态和国内外研究进展等进行介绍。

Tezacaftor/Ivacaftor and Ivacaftor(Symdeko)

由美国Vertex公司研发的复方制剂Symdeko(tezacaftor/ivacaftor和ivacaftor)于2018年2月12日经美国食品与药品管理局(FDA)批准上市,用于跨膜传导调节因子(CFTR)基因携带两个拷贝F508del突变所引起的囊性纤维化的治疗或者至少携带一个对tezacaftor/ivacaftor药物有反应的突变所引起的囊性纤维化的治疗,且适用于年龄在12岁及以上的患者使用[1]。该药为tezacaftor100 mg/ivacaftor 150 mg固定剂量组合片和ivacaftor150 mg片共包装,其中tezacaftor为新化学实体。

囊性纤维化治疗药Ivacaftor

囊性纤维化（CF）是一种严重威胁生命的遗传性疾病,目前全球约有7万患者,其平均寿命仅约为38岁。该病由CF跨膜传导调节蛋白（CFTR）缺陷所引起,CFTR的缺陷或缺失可造成肺部细胞膜上离子流通过量减少,

新型囊性纤维化治疗药Ivacaftor在美国获准上市

美国FDA近期已批准Vertex制药公司开发的首个针对囊性纤维化（CF）病因——cF跨膜转导调节蛋白（CFTR）基因突变的新型口服CF治疗药ivacaftor（Kalydeco）用于CFTR基因发生G55lD突变的6岁或以上年龄的CF患者。CF为一种致死性遗传疾病，可引发慢性肺部感染及进行性肺损伤，并累及其他器官，最终可导致患者过早死亡。

2012年美国FDA批准新药介绍和分析

2012年美国食品药品监督管理局(FDA)共批准39个新药。在2012年FDA批准上市的新药中,有20个为首创一类新药,其中包括首个直接针对病因治疗囊性纤维化的新型药物依伐卡托;40年来首次获准用于多药耐药性肺结核治疗的贝达喹啉;第1个基于FDA的动物有效性研究原则获准上市的新药瑞西巴库单抗;第1个用于艾滋病相关腹泻,也是第1个获准上市的口服植物药可非雷默。简要介绍其中的重点品种,并就新药研发的现状与趋势进行分析。