β-adrenoceptor

Bufuralol (Ro 3-4787) 具有 CYP2D6 底物的芳香环和碱性氮特性，经常用于评估 CYP2D6 活性 [3]。

与心肌中的研究结果一致 [4]，布呋洛尔 (Ro 3-4787) 具有由 NADPH 介导的双相动力学，并且在猴子破坏中的效果不如氢过氧化异丙苯 (CuOOH) 存在时的效果。

细胞色素P450 2D6（CYP2D6）是一种高度多态性的酶，代谢大量治疗药物。迄今为止，已经报道了100多种CYP2D6等位基因变异。在这些变异中，我们最近在中国人群中发现了22种新的变异。本研究的目的是在体外对这些变体的酶活性进行功能表征。杆状病毒介导的表达系统用于高水平表达野生型CYP2D6.1和其他变体（CYP2D6.2、CYP2D6.10和22种新型CYP2D6变体）。然后，将含有表达的CYP2D6蛋白的昆虫微粒体分别与Bufuralol或右美沙芬在37°C下孵育20或25分钟。终止后，提取代谢物并用于高效液相色谱检测。在测试的24个CYP2D6变体中，发现两个变体（CYP2D6.92和CYP2D6.96）没有催化活性。其余22个变体对Bufuralol1'-羟基化的内在清除值显著降低，20个变体对右美沙芬O-去甲基化的内在清理值显著低于野生型CYP2D6.1。我们的体外结果表明，与野生型相比，大多数变体的催化活性显著降低，这些数据为中国和其他亚洲人群的个性化医疗提供了有价值的信息。[2]

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代谢表型可能受到多种因素的影响，包括等位基因变异和与抑制剂的相互作用。人类CYP2D6负责大约20%的细胞色素P450介导的药物代谢，但由100多种已知变体组成；人群中常见几种变体，而其他变体则相当罕见。表达、纯化了四种CYP2D6等位基因变体，其中三种在活性位点远端具有一系列突变（*34、*17-2、*17-3），一种在活性部位附近具有突变的超代谢者（*53），以及参考*1和活性位点突变*1（Thr309Ala），并研究了它们与典型底物右美沙芬和Bufuralol以及灭活剂SCH 66712的相互作用。我们发现*34、*17-2和*17-3在产生与*1相同的代谢产物时，酶活性和NADPH偶联降低，这表明Arg296可能在NADPH偶联中发挥作用。高活性变体*53显示出与*1类似的NADPH偶联，但不易被SCH 66712灭活。Thr309Ala突变体显示出与*1相似的活性，但NADPH偶联大大减少。总体而言，这些结果表明，需要对个体CYP2D6变异进行动力学和代谢分析，以了解它们对可变药物反应的可能贡献以及个性化医疗的复杂性。[3]

(+)-Bufuralol 1'-hydroxylation, a commonly used marker of hepatic CYP2D6 activity, was investigated in human and rhesus monkey intestinal microsomes and compared with that in hepatic microsomes. The cumene hydroperoxide (CuOOH)-mediated metabolism of (+)-bufuralol suggested that at least two enzymes were responsible for bufuralol 1'-hydroxylation in both human and monkey intestinal microsomes. In contrast, the kinetics of the CuOOH-mediated metabolism in human and monkey livers were monophasic. The Km values for the higher affinity component of the intestinal enzyme(s) of both species were similar to, while the corresponding Vmax values were much lower than, those obtained with the livers. Bufuralol metabolism mediated by NADPH exhibited biphasic kinetics and was less efficient than that observed in the presence of CuOOH in both human and monkey intestines, in agreement with the observations in the livers. Inhibition of bufuralol hydroxylase activity in the intestine and liver preparations from the same species by known CYP2D6 inhibitors/substrates was qualitatively similar. Quinidine was the most potent inhibitor of (+)-bufuralol 1'-hydroxylation in all tissues studied. Western immunoblots using anti-CYP2D6 peptide antibody revealed a protein band in human and monkey intestinal microsomes of the same molecular weight as that observed in the liver preparations. The intestinal CYP2D protein content appeared to be much less than that of liver, and correlated with the (+)-bufuralol hydroxylase activity. Immunoinhibition studies indicated significant (up to 50%) inhibition of the CuOOH-mediated (+)-bufuralol metabolism in human and monkey intestines only by anti-CYP2D6, and not by anti-CYP2A6, or anti-CYP2E1. Inhibition of the bufuralol 1'-hydroxylase activity by anti-rat CYP3A1 was only slight (20%) in human, but marked (60-65%) in monkey intestinal microsomes. The hepatic metabolism of (+)-bufuralol in humans and monkeys was only inhibited (75%) by anti-CYP2D6, but not by anti-CYP3A1. Overall, the results suggest that (1) tissue and species differences exist in the catalysis of (+)-bufuralol 1'-hydroxylation, and (2) CYP2D6-related enzymes are partially or primarily responsible for the bufuralol hydroxylase activity in human and monkey intestines or monkey liver[4].

Metabolism / Metabolites
The known human metabolites of ibuprofen include 1'2'-vinyl ibuprofen, 4-hydroxy ibuprofen, and 6-hydroxy ibuprofen.

[1]. Pharmacodynamic and pharmacokinetic studies on bufuralol in man. Br J Clin Pharmacol. 1986 Nov;22(5):527-34.

[2]. Effects of 22 Novel CYP2D6 Variants Found in the Chinese Population on the Bufuralol and Dextromethorphan Metabolisms In Vitro. Basic Clin Pharmacol Toxicol. 2016 Mar;118(3):190-9.

[3]. CYP2D6 Allelic Variants *34, *17-2, *17-3, and *53 and a Thr309Ala Mutant Display Altered Kinetics and NADPH Coupling in Metabolism of Bufuralol and Dextromethorphan and Altered Susceptibility to Inactivation by SCH 66712. Drug Metab Dispos. 2018 Aug;46(8):1106-1117.

[4]. (+)-bufuralol 1'-hydroxylation activity in human and rhesus monkey intestine and liver. Biochem Pharmacol. 1995 Oct 26;50(9):1521-5.

Ibuprofen belongs to the benzofuran class of compounds. Ibuprofen is a novel, non-selective β-adrenergic receptor blocker.

C16H23NO2

261.37

261.173

C, 73.53; H, 8.87; N, 5.36; O, 12.24

54340-62-4

Bufuralol-d9 hydrochloride;1173023-51-2; Bufuralol;54340-62-4; 59652-29-8 (HCl); 60398-91-6 (racemic HCl)

71733

Off-white to light yellow solid powder

1.066g/cm3

393.2ºC at 760 mmHg

191.6ºC

1.558

3.807

45.4

2

3

5

19

287

0

SSEBTPPFLLCUMN-UHFFFAOYSA-N

InChI=1S/C16H23NO2/c1-5-11-7-6-8-12-9-14(19-15(11)12)13(18)10-17-16(2,3)4/h6-9,13,17-18H,5,10H2,1-4H3

2-(tert-butylamino)-1-(7-ethyl-1-benzofuran-2-yl)ethanol

Ro-3-4787; Ro 3-4787; Bufuralolum; Ro 3-4787; Bufuralolum [INN-Latin]; 2-(tert-butylamino)-1-(7-ethyl-1-benzofuran-2-yl)ethanol; 1-(7-Ethylbenzofuran-2-yl)-2-tert-butylamino-1-hydroxyethane; (+/-)-Bufuralol hydrochloride; Bufuralol

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中（例如氮气保护），避免吸湿/受潮和光照。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~382.61 mM)

配方 1 中的溶解度: ≥ 3.5 mg/mL (13.39 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 35.0 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: 3.5 mg/mL (13.39 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
例如，若需制备1 mL的工作液，可将 100 μL 35.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 3.5 mg/mL (13.39 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 35.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。