4-羟基三唑仑 | 65686-11-5

• 分子结构分类: 有机化合物 - 有机杂环化合物 - 苯二氮卓类
• 中文名称: 4-羟基三唑仑
• 英文名称: 4-Hydroxytriazolam
• 英文别名: 1-demethyltriazolam；8-chloro-6-(2-chloro-phenyl)-1-methyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-ol；8-chloro-6-(2-chlorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-ol
• CAS: 65686-11-5
• 化学式: C₁₇H₁₂Cl₂N₄O
• 分子量: 359.214
• InChiKey: YUQRFPYHTPARRM-UHFFFAOYSA-N

物化性质

• 熔点：
  280-282°C
• 溶解度：
  溶于四氢呋喃
• 保留指数：
  3200

计算性质

• 辛醇/水分配系数(LogP)：
  1.9
• 重原子数：
  24
• 可旋转键数：
  1
• 环数：
  4.0
• sp3杂化的碳原子比例：
  0.12
• 拓扑面积：
  63.3
• 氢给体数：
  1
• 氢受体数：
  4

ADMET

代谢

4-羟基三唑安定是已知的triazolam的人类代谢物。

4-Hydroxytriazolam is a known human metabolite of triazolam.

来源:NORMAN Suspect List Exchange

安全信息

• 储存条件：
  -20°C，避光保存。

反应信息

• 作为产物：
  描述：

  三唑仑 在 cynomolgus monkey cytochrome CYP₃A₄ 、 还原型辅酶II(NADPH)四钠盐 作用下， 以 aq. phosphate buffer 为溶剂， 反应 0.25h， 生成 4-羟基三唑仑
  参考文献：
  名称：

  Expression and Characterization of Cynomolgus Monkey Cytochrome CYP3A4 in a Novel Human Embryonic Kidney Cell–Based Mammalian System

  摘要：

  猴是临床前药物发现中常用的物种，其基因与人类高度相似，尤其是药物代谢细胞色素 P450s。然而，在猴与人之间的药物代谢过程中经常会观察到物种差异，要确定这些差异需要表达的 CYPs。为此，研究人员克隆了猴 CYP3A4（c3A4），并在新型人胚肾 293-6E 细胞悬浮系统中进行了表达。制备微粒体后，测定了五种已知人类 CYP3A4（h3A4）底物（咪达唑仑、睾酮、特非那定、硝苯地平和三唑仑）的动力学特征。尽管 K m 值存在一些差异，但所有五种底物都是 c3A4 的良好底物。总体而言，数据表明 c3A4 和 h3A4 底物具有很强的相似性。此外，除了已知的人类 CYP2D6 底物（糠醛醇）外，c3A4 对非 h3A4 探针底物没有表现出活性，这表明 c3A4 对人类细胞色素 CYP2D6 底物有潜在的代谢作用。酮康唑和曲安奈德霉素对 c3A4 和 h3A4 的抑制效力相似，而非 h3A4 抑制剂对 c3A4 活性没有抑制作用。c3A4制剂和市售猴肝微粒体的出现将有助于进一步确定猴作为模型的特性，以评估CYP3A依赖性清除和药物相互作用。

  DOI：

  10.1124/dmd.113.055491

文献信息

• Metabolic Disposition of Triazolam and Clobazam in Humanized CYP3A Mice with a Double-Knockout Background of Mouse<i>Cyp2c</i>and<i>Cyp3a</i>Genes
  作者：Kaoru Kobayashi、Genki Minegishi、Nina Kuriyama、Atsushi Miyajima、Satoshi Abe、Kanako Kazuki、Yasuhiro Kazuki

  DOI：10.1124/dmd.122.001087

  日期：2023.2

  Knockout (KO) of mouse Cyp3a genes increases the expression of hepatic CYP2C enzymes, which can metabolize triazolam, a typical substrate of human CYP3A. There is still marked formation of 1’-hydroxytriazolam in Cyp3a-KO (3aKO) mice after triazolam dosing. Here, we generated a new model of humanized CYP3A (hCYP3A) mice with a double-KO background of Cyp3a and Cyp2c genes (2c3aKO), and we examined the metabolic profiles of triazolam in wild-type (WT), 2c3aKO, and hCYP3A/2c3aKO mice in vitro and in vivo . In vitro studies using liver microsomes showed that the formation of 1’-hydroxytriazolam in 2c3aKO mice was less than 8% of that in WT mice. The formation rate of 1’-hydroxytriazolam in hCYP3A/2c3aKO mice was eightfold higher than that in 2c3aKO mice. In vivo studies showed that area under the curve (AUC) of 1’-hydroxytriazolam in 2c3aKO mice was less than 3% of that in WT mice. The AUC of 1’-hydroxytriazolam in hCYP3A/2c3aKO mice was sixfold higher than that in 2c3aKO mice. These results showed that formation of 1’-hydroxytriazolam was significantly decreased in 2c3aKO mice. Metabolic functions of human CYP3A enzymes were distinctly found in hCYP3A mice with the 2c3aKO background. Moreover, hCYP3A/2c3aKO mice treated with clobazam showed human CYP3A-mediated formation of desmethylclobazam and prolonged elimination of desmethylclobazam, which is found in poor metabolizers of CYP2C19. The novel hCYP3A mouse model without mouse Cyp2c and Cyp3a genes (hCYP3A/2c3aKO) is expected to be useful to evaluate human CYP3A-mediated metabolism in vivo . SIGNIFICANT STATEMENT Humanized CYP3A (hCYP3A/2c3aKO) mice with a background of double knockout (KO) for mouse Cyp2c and Cyp3a genes were generated. Although CYP2C enzymes played a compensatory role in the metabolism of triazolam to 1’-hydroxytriazolam in the previous hCYP3A/3aKO mice with Cyp2c genes, the novel hCYP3A/2c3aKO mice clearly showed functions of human CYP3A enzymes introduced by chromosome engineering technology.

  小鼠 Cyp3a 基因敲除（KO）会增加肝脏 CYP2C 酶的表达量，而 CYP2C 酶可以代谢人类 CYP3A 的典型底物--三唑仑。在服用三唑仑后，Cyp3a-KO（3aKO）小鼠体内仍有明显的1'-羟基三唑仑形成。在这里，我们生成了一种具有 Cyp3a 和 Cyp2c 基因双 KO（2c3aKO）背景的人源化 CYP3A（hCYP3A）小鼠新模型，并研究了三唑仑在野生型（WT）、2c3aKO 和 hCYP3A/2c3aKO 小鼠体内和体外的代谢情况。使用肝脏微粒体进行的体外研究表明，2c3aKO 小鼠体内 1'-hydroxytriazolam 的形成率不到 WT 小鼠的 8%。hCYP3A/2c3aKO 小鼠体内 1'-hydroxytriazolam 的形成率是 2c3aKO 小鼠的八倍。体内研究表明，2c3aKO 小鼠体内 1'-hydroxytriazolam 的曲线下面积（AUC）不到 WT 小鼠的 3%。hCYP3A/2c3aKO 小鼠体内 1'-hydroxytriazolam 的 AUC 是 2c3aKO 小鼠的六倍。这些结果表明，在 2c3aKO 小鼠体内，1'-羟基三唑仑的形成明显减少。在具有 2c3aKO 背景的 hCYP3A 小鼠中，人类 CYP3A 酶的代谢功能被明显发现。此外，hCYP3A/2c3aKO小鼠接受氯巴扎姆治疗后，显示出人类CYP3A介导的去甲基氯巴扎姆的形成和去甲基氯巴扎姆消除时间的延长，这在CYP2C19代谢不良的小鼠中是可以发现的。没有小鼠 Cyp2c 和 Cyp3a 基因的新型 hCYP3A 小鼠模型（hCYP3A/2c3aKO）有望用于评估人类 CYP3A 介导的体内代谢。 重要声明 生成了以小鼠 Cyp2c 和 Cyp3a 基因双基因敲除（KO）为背景的人源化 CYP3A（hCYP3A/2c3aKO）小鼠。虽然在以前的带有 Cyp2c 基因的 hCYP3A/3aKO 小鼠中，CYP2C 酶在将三唑仑代谢为 1'-hydroxytriazolam 的过程中发挥了补偿作用，但新型 hCYP3A/2c3aKO 小鼠明确显示了通过染色体工程技术引入的人 CYP3A 酶的功能。
• Prediction of metabolic clearance using fresh human hepatocytes: Comparison with cryopreserved hepatocytes and hepatic microsomes for five benzodiazepines
  作者：D. Hallifax、A. Galetin、J. B. Houston

  DOI：10.1080/00498250701834665

  日期：2008.4

  1. Predictions of in vivo intrinsic clearance from cryopreserved human hepatocytes may be systematically low. In the current study, the metabolite kinetics of a series of CYP3A4 substrates (benzodiazepines) in fresh human hepatocytes from five donors, via a major UK supplier, were investigated and compared with those previously reported (by the authors' laboratory) for cryopreserved human hepatocytes and hepatic microsomes.2. A high incidence of autoactivation (up to tenfold) and heteroactivation (by testosterone, up to 14-fold) among the major pathways was observed. CYP capacity (V-max) was marginally lower and 'affinity' constants (K-M, S-50) were marginally greater compared with cryopreserved hepatocytes.3. Average intrinsic clearance (based on maximal clearance, CLmax) was sevenfold lower than in cryopreserved hepatocytes (reflecting sensitivity of intrinsic clearance estimation in vitro to mechanistic parameter values, particularly those involving atypical kinetics), but scaled intrinsic clearances for fresh (and cryopreserved) hepatocytes were within the range previously determined in hepatic microsomes.4. There was no evidence from this series of studies that fresh hepatocytes provide quantitatively improved estimates of intrinsic clearance over cryopreserved hepatocytes.
• Expression and Characterization of Cynomolgus Monkey Cytochrome CYP3A4 in a Novel Human Embryonic Kidney Cell–Based Mammalian System
  作者：Sindhuja Selvakumar、Priyadeep Bhutani、Kaushik Ghosh、Prasad Krishnamurthy、Sanjith Kallipatti、Sabariya Selvam、Manjunath Ramarao、Sandhya Mandlekar、Michael W. Sinz、A. David Rodrigues、Murali Subramanian

  DOI：10.1124/dmd.113.055491

  日期：2014.3

  Cynomolgus monkeys are a commonly used species in preclinical drug discovery, and have high genetic similarity to humans, especially for the drug-metabolizing cytochrome P450s. However, species differences are frequently observed in the metabolism of drugs between cynomolgus monkeys and humans, and delineating these differences requires expressed CYPs. Toward this end, cynomolgus monkey CYP3A4 (c3A4) was cloned and expressed in a novel human embryonic kidney 293-6E cell suspension system. Following the preparation of microsomes, the kinetic profiles of five known human CYP3A4 (h3A4) substrates (midazolam, testosterone, terfenadine, nifedipine, and triazolam) were determined. All five substrates were found to be good substrates of c3A4, although some differences were observed in the K m values. Overall, the data suggest a strong substrate similarity between c3A4 and h3A4. Additionally, c3A4 exhibited no activity against non-h3A4 probe substrates, except for a known human CYP2D6 substrate (bufuralol), which suggests potential metabolism of human cytochrome CYP2D6-substrates by c3A4. Ketoconazole and troleandomycin showed similar inhibitory potencies toward c3A4 and h3A4, whereas non-h3A4 inhibitors did not inhibit c3A4 activity. The availability of a c3A4 preparation, in conjunction with commercially available monkey liver microsomes, will support further characterization of the cynomolgus monkey as a model to assess CYP3A-dependent clearance and drug-drug interactions.

  猴是临床前药物发现中常用的物种，其基因与人类高度相似，尤其是药物代谢细胞色素 P450s。然而，在猴与人之间的药物代谢过程中经常会观察到物种差异，要确定这些差异需要表达的 CYPs。为此，研究人员克隆了猴 CYP3A4（c3A4），并在新型人胚肾 293-6E 细胞悬浮系统中进行了表达。制备微粒体后，测定了五种已知人类 CYP3A4（h3A4）底物（咪达唑仑、睾酮、特非那定、硝苯地平和三唑仑）的动力学特征。尽管 K m 值存在一些差异，但所有五种底物都是 c3A4 的良好底物。总体而言，数据表明 c3A4 和 h3A4 底物具有很强的相似性。此外，除了已知的人类 CYP2D6 底物（糠醛醇）外，c3A4 对非 h3A4 探针底物没有表现出活性，这表明 c3A4 对人类细胞色素 CYP2D6 底物有潜在的代谢作用。酮康唑和曲安奈德霉素对 c3A4 和 h3A4 的抑制效力相似，而非 h3A4 抑制剂对 c3A4 活性没有抑制作用。c3A4制剂和市售猴肝微粒体的出现将有助于进一步确定猴作为模型的特性，以评估CYP3A依赖性清除和药物相互作用。