1α,23(S),25-trihydroxyvitamin D3 | 86701-33-9

• 分子结构分类: 有机化合物 - 脂质和类脂质分子 - 类固醇和类固醇衍生物
• 英文名称: 1α,23(S),25-trihydroxyvitamin D3
• 英文别名: 1,23S, 25D₃；1alpha,23(S),25-trihydroxyvitamin D3；(1R,3S,5Z)-5-[(2E)-2-[(1R,3aS,7aR)-1-[(2R,4S)-4,6-dihydroxy-6-methylheptan-2-yl]-7a-methyl-2,3,3a,5,6,7-hexahydro-1H-inden-4-ylidene]ethylidene]-4-methylidenecyclohexane-1,3-diol
• CAS: 86701-33-9
• 化学式: C₂₇H₄₄O₄
• 分子量: 432.644
• InChiKey: NHRGJVVEKNHIIE-KWJCNBFNSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  3.9
• 重原子数：
  31
• 可旋转键数：
  6
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.78
• 拓扑面积：
  80.9
• 氢给体数：
  4
• 氢受体数：
  4

反应信息

• 作为产物：
  描述：

  (8S,20R)-de-A,B-8-(tert-butyldimethylsilyl)oxy-20-(formylmethyl)pregnane 在 4-二甲氨基吡啶 、 indium 、 正丁基锂 、 四丙基高钌酸铵 、 氢氟酸 、 水 、 对甲苯磺酸 、 N-甲基吗啉氧化物 、 三乙胺 作用下， 以 四氢呋喃 、 甲醇 、 二氯甲烷 、 乙腈 为溶剂， 生成 1α,23(S),25-trihydroxyvitamin D3
  参考文献：
  名称：

  CYP27B1化学合成侧链羟基化维生素D3衍生物及其代谢

  摘要：

  研究了 CYP27B1在一系列侧链羟基化 D 3衍生物的 C1 羟基化中的作用。我们发现与相应的S非对映异构体相比，24 R和 26 R代谢物在 C1 处被 CYP27B1 更有效地羟基化。然而，CYP27B1 对 23-羟基化衍生物的任何一种非对映异构体几乎没有表现出活性。这是第一份表明 CYP27B1 代谢 26-羟基化 D 3，将 25,26D 3转化为相应的 1α-羟基化化合物 1,25,26D 3的报告。

  DOI：

  10.1002/cbic.202100250

文献信息

• Single A326G mutation converts human CYP24A1 from 25-OH-D ₃ -24-hydroxylase into -23-hydroxylase, generating 1α,25-(OH) ₂ D ₃ -26,23-lactone
  作者：David E. Prosser、Martin Kaufmann、Brendan O'Leary、Valarie Byford、Glenville Jones

  DOI：10.1073/pnas.0702093104

  日期：2007.7.31

  Studies of 25-hydroxyvitamin D ₃ -24-hydroxylase (CYP24A1) have demonstrated that it is a bifunctional enzyme capable of the 24-hydroxylation of 1α,25-(OH) ₂ D ₃ , leading to the excretory form, calcitroic acid, and 23-hydroxylation, culminating in 1α,25-(OH) ₂ D ₃ -26,23-lactone. The degree to which CYP24A1 performs either 23- or 24-hydroxylation is species-dependent. In this paper, we show that the human enzyme that predominantly 24-hydroxylates its substrate differs from the opossum enzyme that 23-hydroxylates it at only a limited number of amino acid residues. Mutagenesis of the human form at a single substrate-binding residue (A326G) dramatically changes the regioselectivity of the enzyme from a 24-hydroxylase to a 23-hydroxylase, whereas other modifications have no effect. Ala-326 is located in the I-helix, close to the terminus of the docked 25-hydroxylated side chain in a CYP24A1 homology model, a result that we interpret indicates that substitution of a glycine at 326 provides extra space for the side chain of the substrate to move deeper into the pocket and place it in a optimal stereochemical position for 23-hydroxylation. We discuss the physiological ramifications of these results for species possessing the A326G substitution, as well as implications for optimal vitamin D analog design.

  对25-羟基维生素D3-24-羟化酶（CYP24A1）的研究表明，它是一种具有双重功能的酶，能够对1α，25-（OH）2D3进行24-羟基化，形成排泄形式的钙三酸，以及23-羟基化，最终形成1α，25-（OH）2D3-26,23-内酯。CYP24A1执行23-或24-羟基化的程度因物种而异。在本文中，我们展示了主要对其底物进行24-羟基化的人类酶与仅在有限数量的氨基酸残基上对其进行23-羟基化的负鼠酶有所不同。通过对人类形式的单个底物结合残基（A326G）进行突变，可以将酶的区域选择性从24-羟基化酶显著地改变为23-羟基化酶，而其他修饰则没有影响。Ala-326位于I-螺旋中，靠近CYP24A1同源模型中停靠的25-羟基化侧链的末端，我们解释这个结果表明在326处替换甘氨酸提供了额外的空间，使底物的侧链更深地进入袋中，并将其放置在最佳立体化学位置进行23-羟基化。我们讨论了这些结果对具有A326G突变的物种的生理影响，以及对最佳维生素D类似物设计的影响。
• Structure-Function Analysis of Vitamin D 24-Hydroxylase (CYP24A1) by Site-Directed Mutagenesis: Amino Acid Residues Responsible for Species-Based Difference of CYP24A1 between Humans and Rats
  作者：Hiromi Hamamoto、Tatsuya Kusudo、Naoko Urushino、Hiroyuki Masuno、Keiko Yamamoto、Sachiko Yamada、Masaki Kamakura、Miho Ohta、Kuniyo Inouye、Toshiyuki Sakaki

  DOI：10.1124/mol.106.023275

  日期：2006.7

  Our previous studies revealed the species-based difference of CYP24A1-dependent vitamin D metabolism. Although human CYP24A1 catalyzes both C-23 and C-24 oxidation pathways, rat CYP24A1 shows almost no C-23 oxidation pathway. We tried to identify amino acid residues that cause the species-based difference by site-directed mutagenesis. In the putative substrate-binding regions, amino acid residue of rat CYP24A1 was converted to the corresponding residue of human CYP24A1. Among eight mutants examined, T416M and I500T showed C-23 oxidation pathway. In addition, the mutant I500F showed quite a different metabolism of 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3] from both human and rat CYP24A1. These results strongly suggest that the amino acid residues at positions 416 and 500 play a crucial role in substrate binding and greatly affect substrate orientation. A three-dimensional model of CYP24A1 indicated that the A-ring and triene part of 1α,25(OH)2D3 could be located close to amino acid residues at positions 416 and 500, respectively. Our findings provide useful information for the development of new vitamin D analogs for clinical use.

  我们之前的研究揭示了CYP24A1依赖性维生素D代谢的物种差异。虽然人类CYP24A1催化C-23和C-24氧化途径，但大鼠CYP24A1几乎没有C-23氧化途径。我们试图通过定点突变来确定导致物种差异的氨基酸残基。在推测的底物结合区域，大鼠CYP24A1的氨基酸残基被转化为人类CYP24A1的相应残基。在所研究的八个突变体中，T416M和I500T表现出C-23氧化途径。此外，突变体I500F表现出与人类和大鼠CYP24A1截然不同的1α,25-二羟基维生素D3（1α,25(OH)2D3）代谢。这些结果有力地表明，第416和500位的氨基酸残基在底物结合中起着至关重要的作用，并极大地影响底物取向。CYP24A1的三维模型表明，1α,25(OH)2D3的A环和三烯部分可能分别位于靠近第416和500位的氨基酸残基的位置。我们的发现为开发新的维生素D类似物用于临床提供了有用的信息。
• Enhanced biosynthesis of physiologically active vitamin D3 by constructing recombinant Escherichia coli BL21 with a multienzyme system
  作者：Bing Fu、Liuzhen Yang、Qingwei Chen、Qili Zhang、Lei Zhang、Ping Yu

  DOI：10.1016/j.procbio.2022.12.026

  日期：2023.2

  In this study, a recombinant E. coli strain with the multienzyme system of the vitamin D3 hydroxylase CYP105A1, the electron transport respiratory chain Fdx-FdR and the key enzymes (Glk-Zwf) in the cyclic regeneration of the coenzyme NADPH was constructed for the efficient hydroxylation of VD3 to prepare physiologically active 25(OH)VD3 and 1α, 25(OH)2VD3. The constructed recombinant E. coli strain

  在本研究中，构建了具有维生素 D 3羟化酶 CYP105A1、电子传递呼吸链 Fdx-FdR 和辅酶 NADPH 循环再生中的关键酶 (Glk-Zwf)的多酶系统的重组大肠杆菌菌株，用于VD 3高效羟基化制备具有生理活性的25(OH)VD 3和1α, 25(OH) 2 VD 3。构建的具有多酶系统的重组大肠杆菌菌株被证明具有对VD 3的羟基化能力。VD 3的羟基化反应是在最佳条件下进行的。中间产物25(OH)VD 3的最高浓度为58.92 mg/L，产率为11.31 %。最终产物1α, 25(OH) 2 VD 3的最高浓度为90.23 mg/L，产率为16.7%。最终，通过进一步的底物补料分批实验产生了395.92 mg/L 25(OH)VD 3和 272.21 mg/L 1α, 25(OH) 2 VD 3 。本研究有助于通过构建的重组菌株对VD 3进行全细胞生物催化制备具有生理活性的25(OH)VD
• Kinetic analysis of human CYP24A1 metabolism of vitamin D via the C24-oxidation pathway
  作者：Elaine W. Tieu、Edith K. Y. Tang、Robert C. Tuckey

  DOI：10.1111/febs.12862

  日期：2014.7

  CYP24A1 is the multicatalytic cytochrome P450 responsible for the catabolism of vitamin D via the C23‐ and C24‐oxidation pathways. We successfully expressed the labile human enzyme in Escherichia coli and partially purified it in an active state that permitted detailed characterization of its metabolism of 1,25‐dihydroxyvitamin D3 [1,25(OH)2D3] and the intermediates of the C24‐oxidation pathway in a phospholipid‐vesicle reconstituted system. The C24‐oxidation pathway intermediates, 1,24,25‐trihydroxyvitamin D3, 24‐oxo‐1,25‐dihydroxyvitamin D3, 24‐oxo‐1,23,25‐trihydroxyvitamin D3 and tetranor‐1,23‐dihydroxyvitamin D3, were enzymatically produced from 1,25(OH)2D3 using rat CYP24A1. Both 1,25(OH)2D3 and 1,23‐dihydroxy‐24,25,26,27‐tetranorvitamin D3 were found to partition strongly into the phospholipid bilayer when in aqueous medium. Changes to the phospholipid concentration did not affect the kinetic parameters for the metabolism of 1,25(OH)2D3 by CYP24A1, indicating that it is the concentration of substrates in the membrane phase (mol substrate·mol phospholipid−1) that determines their rate of metabolism. CYP24A1 exhibited Km values for the different C24‐intermediates ranging from 0.34 to 15 mmol·mol phospholipid−1, with 24‐oxo‐1,23,25‐trihydroxyvitamin D3 [24‐oxo‐1,23,25(OH)3D3] displaying the lowest and 1,24,25‐trihydroxyvitamin D3 [1,24,25(OH)3D3] displaying the highest. The kcat values varied by up to 3.8‐fold, with 1,24,25(OH)3D3 displaying the highest kcat (34 min−1) and 24‐oxo‐1,23,25(OH)3D3 the lowest. The data show that the cleavage of the side chain of 24‐oxo‐1,23,25(OH)3D3 occurs with the highest catalytic efficiency (kcat/Km) and produces 1‐hydroxy‐23‐oxo‐24,25,26,27‐tetranorvitamin D3 and not 1,23‐dihydroxy‐24,25,26,27‐tetranorvitamin D3, as the primary product. These kinetic analyses also show that intermediates of the C24‐oxidation pathway effectively compete with precursor substrates for binding to the active site of the enzyme, which manifests as an accumulation of intermediates, indicating that they dissociate after each catalytic step.
• Chemical Synthesis of Side‐Chain‐Hydroxylated Vitamin D ₃ Derivatives and Their Metabolism by CYP27B1
  作者：Ryota Sakamoto、Akiko Nagata、Haruki Ohshita、Yuka Mizumoto、Miho Iwaki、Kaori Yasuda、Toshiyuki Sakaki、Kazuo Nagasawa

  DOI：10.1002/cbic.202100250

  日期：2021.10

  side-chain-hydroxylated D3 derivatives was explored. We found that the 24R and 26R metabolites were more effectively hydroxylated at C1 by CYP27B1 compared to the corresponding S diastereomers. However, CYP27B1 showed almost no activity towards either of the diastereomers of the 23-hydroxylated derivative. This is the first report to show that CYP27B1 metabolizes 26-hydroxylated D3, converting 25,26D3

  研究了 CYP27B1在一系列侧链羟基化 D 3衍生物的 C1 羟基化中的作用。我们发现与相应的S非对映异构体相比，24 R和 26 R代谢物在 C1 处被 CYP27B1 更有效地羟基化。然而，CYP27B1 对 23-羟基化衍生物的任何一种非对映异构体几乎没有表现出活性。这是第一份表明 CYP27B1 代谢 26-羟基化 D 3，将 25,26D 3转化为相应的 1α-羟基化化合物 1,25,26D 3的报告。