Moscow Research and Practical Centre on Addictions of the Moscow Department of Healthcare, 37/1 Lyublinskaya Street, Moscow 109390, Russia; Medical Academy of Continuous Professional Education of the Ministry of Health of the Russian Federation, 2/1 Barrikadnaya Street, Moscow 123995, Russian Federation.
Moscow Research and Practical Centre on Addictions of the Moscow Department of Healthcare, 37/1 Lyublinskaya Street, Moscow 109390, Russia.
Gene. 2020 May 20;739:144513. doi: 10.1016/j.gene.2020.144513. Epub 2020 Feb 26.
Alprazolam is used in the treatment of patients with anxiety disorders comorbid with alcohol use disorder. Some proportion of these patients does not respond adequately to treatment with alprazolam, while many of them experience dose-dependent adverse drug reactions. Results of the previous studies have shown that CYP3A is involved in the biotransformation of alprazolam, the activity of which is dependent, inter alia, on the polymorphism of the encoding gene.
The objective of our study was to investigate the effect of 99366316G>A polymorphism of the CYP3A4 gene on the concentration/dose indicator of alprazolam in patients with anxiety disorders comorbid with alcohol use disorder, using findings on enzymatic activity of CYP3A (as evaluated by the 6-beta-hydroxy-cortisol/cortisol ratio measurement) and on CYP3A4 expression level obtained by measuring the miR-27b plasma concentration levels in patients with anxiety disorders comorbid with alcoholism.
Our study enrolled 105 patients with anxiety disorders comorbid with alcohol use disorder (age - 37.8±14.6 years). Therapy included alprazolam in an average daily dose of 5.6±2.4 mg per day. Treatment efficacy was evaluated using the psychometric scales. Therapy safety was assessed using the UKU Side-Effect Rating Scale. For genotyping and estimation of the microRNA (miRNA) plasma levels, we performed the real-time polymerase chain reaction. The activity of CYP3A was evaluated using the HPLC-MS/MS method by the content of the endogenous substrate of the given isoenzyme and its metabolite in urine (6- beta-hydroxy-cortisol/cortisol). Therapeutic drug monitoring (TDM) has been performed using HPLC-MS/MS.
Our study revealed the statistically significant results in terms of the treatment efficacy evaluation (HAMA scores at the end of the treatment course): (GG) 3.0 [2.0; 5.0] and (GA) 4.0 [4.0; 5.0], p = 0.007; at the same time, the statistical significance in the safety profile was not obtained (the UKU scores): (GG) 3.0 [2.0; 3.8] and (GA) 3.0 [1.5; 4.0], p = 0.650. We revealed a statistical significance for concentration/dose indicator of alprazolam in patients with different genotypes: (GG) 1.583 [0.941; 2.301] and (GA) 2.888 [2.305; 4.394], p = 0.001). Analysis of the results of the pharmacotranscriptomic part of the study didn't show the statistically significant difference in the miR-27b plasma levels in patients with different genotypes: (GG) 25.6 [20.4; 28.8], (GA) 25.7 [19.7; 33.1], p = 0.423. At the same time, correlation analysis revealed a statistically significant relationship between the alprazolam efficacy profile evaluated by changes in HAMA scale scores and the miR-27b plasma concentration: rs = 0.20, p = 0.042. Also, we didn't reveal the correlation between the miRNA concentration and safety profile: rs = 0.15, p = 0.127. In addition, we revealed the relationship between the CYP3A enzymatic activity (as evaluated by 6-beta-hydroxycortisol/ cortisol ratio measurement) and the miR-27b plasma concentration: rs = -0.27, p = 0.006. At the same time, correlation analysis revealed a statistically significant relationship between the alprazolam concentration and the miR-27b plasma concentration: rs = 0.28, p = 0.003.
The effect of genetic polymorphism of the CYP3A4 gene on the efficacy and safety profiles of alprazolam was demonstrated in a group of 105 patients with anxiety disorders comorbid with alcohol use disorder. At the same time, miR-27b remains a promising biomarker for assessing the level of CYP3A4 expression, because it correlates with the encoded isoenzyme's activity.
阿普唑仑用于治疗伴有酒精使用障碍的焦虑症患者。这些患者中有一定比例对阿普唑仑的治疗反应不足，而他们中的许多人则经历剂量依赖性的药物不良反应。先前的研究结果表明，CYP3A 参与了阿普唑仑的生物转化，其活性除其他外取决于编码基因的多态性。
我们的研究目的是通过测量焦虑症伴酒精使用障碍患者的 CYP3A 酶活性（通过 6-β-羟基皮质醇/皮质醇比值测量评估）和 miR-27b 血浆浓度水平，研究 CYP3A4 基因的 99366316G>A 多态性对伴有酒精使用障碍的焦虑症患者的阿普唑仑浓度/剂量指标的影响。
我们纳入了 105 例伴有酒精使用障碍的焦虑症患者（年龄 37.8±14.6 岁）。治疗包括阿普唑仑，平均每日剂量为 5.6±2.4mg/天。使用心理计量学量表评估治疗效果，使用 UKU 副作用评定量表评估治疗安全性。通过实时聚合酶链反应进行基因分型和微 RNA (miRNA) 血浆水平的测定。使用 HPLC-MS/MS 方法通过内源性同工酶及其代谢物在尿液中的含量（6-β-羟基皮质醇/皮质醇）评估 CYP3A 活性。使用 HPLC-MS/MS 进行治疗药物监测（TDM）。
我们的研究揭示了在治疗效果评估方面具有统计学意义的结果（治疗过程结束时的 HAMA 评分）：（GG）3.0[2.0；5.0]和（GA）4.0[4.0；5.0]，p=0.007；同时，在安全性方面未获得统计学意义（UKU 评分）：（GG）3.0[2.0；3.8]和（GA）3.0[1.5；4.0]，p=0.650。我们发现不同基因型的患者的阿普唑仑浓度/剂量指标存在统计学意义：（GG）1.583[0.941；2.301]和（GA）2.888[2.305；4.394]，p=0.001。对研究中药物转录组学部分的结果分析显示，不同基因型患者的 miR-27b 血浆水平没有统计学差异：（GG）25.6[20.4；28.8]，（GA）25.7[19.7；33.1]，p=0.423。同时，相关分析显示，HAMA 评分变化评估的阿普唑仑疗效与 miR-27b 血浆浓度之间存在统计学关系：rs=0.20，p=0.042。此外，我们没有发现 miRNA 浓度与安全性之间的相关性：rs=0.15，p=0.127。另外，我们还发现了 CYP3A 酶活性（通过 6-β-羟基皮质醇/皮质醇比值测量评估）与 miR-27b 血浆浓度之间的关系：rs=-0.27，p=0.006。同时，相关分析显示，阿普唑仑浓度与 miR-27b 血浆浓度之间存在统计学关系：rs=0.28，p=0.003。
在伴有酒精使用障碍的焦虑症患者组中，证明了 CYP3A4 基因的遗传多态性对阿普唑仑的疗效和安全性的影响。同时，miR-27b 仍然是评估 CYP3A4 表达水平的有前途的生物标志物，因为它与编码同工酶的活性相关。
