1,25-二羟基-24-氧代-维他命D3 | 76338-50-6

• 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
• 中文名称: 1,25-二羟基-24-氧代-维他命D3
• 英文名称: 1α,25-dihydroxy-24-oxovitamin D₃
• 英文别名: 1α,25-dihydroxy-24-oxovitamin-D₃；24-oxo-1,25-dihydroxyvitamin D₃；1α,25-dihydroxy-24-oxo-vitamin D3；1α,25-(OH)2-24-oxo-D₃；1α,25-dihydroxy-24-oxovitamin D3；24-oxo-1,25-dihydroxyvitamin D3；(1S)-1,25-dihydroxy-24-oxocalciol；(6R)-6-[(1R,3aS,4E,7aR)-4-[(2Z)-2-[(3S,5R)-3,5-dihydroxy-2-methylidenecyclohexylidene]ethylidene]-7a-methyl-2,3,3a,5,6,7-hexahydro-1H-inden-1-yl]-2-hydroxy-2-methylheptan-3-one
• CAS: 76338-50-6
• 化学式: C₂₇H₄₂O₄
• 分子量: 430.628
• InChiKey: BWFQMABKLLTETH-YGQRWWDYSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  3.8
• 重原子数：
  31
• 可旋转键数：
  6
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.74
• 拓扑面积：
  77.8
• 氢给体数：
  3
• 氢受体数：
  4

反应信息

• 作为反应物：
  描述：

  1,25-二羟基-24-氧代-维他命D3 、 氢(+1)阳离子 、 氧气 、 [2Fe-2S]¹⁺ 生成 (1S,3R,5Z,7E)-1,3,23,25-四羟基-9,10-开链胆甾-5,7,10-三烯-24-酮 、 水 、 [2Fe-2S]²⁺
  参考文献：
  名称：

  涉及维生素D活化和失活的酶。

  摘要：

  已显示六种细胞色素P450（CYP）亚型可羟化维生素D。其中四个，CYP27A1，CYP2R1，CYP3A4和CYP2J3，是参与活化第一步的维生素D 25-羟化酶的候选物。高度管制的肾脏酶25-羟基维生素D-1α-羟化酶包含CYP27B1成分，该成分完成了向激素形式1α，25-二羟基维生素D（3）的激活途径。从1alpha，25-（OH）（2）D（3）到钙柠檬酸的五步失活途径归因于单个多功能CYP CYP24A1，它通过1alpha，25在维生素D目标细胞中转录诱导-（OH）（2）D（3）。根据其他CYP的排列和晶体结构，已经生成了维生素D相关CYP的同源性模型。

  DOI：

  10.1016/j.tibs.2004.10.005
• 作为产物：
  描述：

  骨化三醇 在 bovine adrenodoxin reductase 、 bovine adrenodoxin 、 recombinant rat cytochrome P450 24A1(Δ2-32) 、 还原型辅酶II(NADPH)四钠盐 作用下， 反应 0.08h， 生成 (3R,5S)-5-[(2R)-2-[(1R,3aR,4E,7aR)-4-[(2Z)-2-[(3S,5S)-3,5-二羟基-2-亚甲基-环己基亚基]乙亚基]-7a-甲基-2,3,3a,5,6,7-六氢-1H-茚-1-基]丙基]-3-羟基-3-甲基-四氢呋喃-2-酮 、 1,23-二羟基-24,25,26,27-四去甲维他命 D3 、 1,25-二羟基-24-氧代-维他命D3 、 (24R)-1(alpha),24,25-三羟基维他命 D3* 、 1α,23(S),25-trihydroxy-24-oxovitamin D₃
  参考文献：
  名称：

  A new insight into the role of rat cytochrome P450 24A1 in metabolism of selective analogs of 1α,25-dihydroxyvitamin D3

  摘要：

  We examined the metabolism of two synthetic analogs of 1 alpha,25-dihydroxyvitamin D-3 (1), namely 1 alpha,25-dihydroxy-16-ene-23-yne-vitamin D-3 (2) and 1 alpha,25-dihydroxy-16-ene-23-yne-26,27-dimethyl-vitamin D-3 (4) using rat cytochrome P450 24A1 (CYP24A1) in a reconstituted system. We noted that 2 is metabolized into a single metabolite identified as C26-hydroxy-2 while 4 is metabolized into two metabolites, identified as C26-hydroxy-4 and C26a-hydroxy-4. The structural modification of adding methyl groups to the side chain of 1 as in 4 is also featured in another analog, 1 alpha,25-dihydroxy-22,24-diene-24,26,27-tri-homo-vitamin D-3 (6). In a previous study, 6 was shown to be metabolized exactly like 4, however, the enzyme responsible for its metabolism was found to be not CYP24A1. To gain a better insight into the structural determinants for substrate recognition of different analogs, we performed an in silico docking analysis using the crystal structure of rat CYP24A1 that had been solved for the substrate-free open form. Whereas analogs 2 and 4 docked similar to 1,6 showed altered interactions for both the A-ring and side chain, despite prototypical recognition of the CD-ring. These findings hint that CYP24A1 metabolizes selectively different analogs of 1, based on their ability to generate discrete recognition cues required to close the enzyme and trigger the catalytic mechanism. 2011 Elsevier Inc. All rights reserved.

  DOI：

  10.1016/j.abb.2011.02.004

文献信息

• 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。
• Composition for injection of active type vitamins D3
  申请人：TEIJIN LIMITED

  公开号：EP0215596A2

  公开（公告）日：1987-03-25

  A composition for the injection of active type vitamins D₃ comprising a lyophilized product of active type vitamins D₃ and an excipient. This composition has good stability of active type vitamins D₃ and is effective as an injection preparation. This composition can be easily prepared.

  一种用于注射活性维生素 D₃ 的组合物，由活性维生素 D₃ 的冻干产品和赋形剂组成。 这种组合物具有良好的活性维生素 D₃ 稳定性，作为注射制剂效果显著。 这种组合物易于制备。
• MEDICINE-CONTAINING FAT EMULSION OF THE TYPE PREPARED IMMEDIATELY BEFORE USE AND PROCESS FOR PREPARING MEDICINE-CONTAINING FAT EMULSION
  申请人：TEIJIN LIMITED

  公开号：EP0331755A1

  公开（公告）日：1989-09-13

  A kit of a medicine-containing fat emulsion of the type prepared immediately before its use, which comprises a fat emulsion and either (a) a medicinal composition comprising a medicine and at least one solvent selected from among li­quid polyalkylene glycols, liquid alkylethanolamines, liquid polyhydric alcohols and water, or (b) a medicinal composition comprising a medicine and an excipient such as a sugar and/or an amino acid, and a process for preparing a medicine-­containing fat emulsion therefrom immediately before its use.

  一种在使用前立即制备的含药脂肪乳剂试剂盒，它包括脂肪乳剂和(a)由药物和至少一种选自液态聚烷乙二醇、液态烷基乙醇胺、液态多羟基乙醇和水的溶剂组成的药物组合物，或(b)由药物和糖和/或氨基酸等赋形剂组成的药物组合物，以及在使用前立即制备含药脂肪乳剂的工艺。
• Expression of human CYP27B1 in<i>Escherichia coli</i>and characterization in phospholipid vesicles
  作者：Edith K. Y. Tang、Elaine W. Tieu、Robert C. Tuckey

  DOI：10.1111/j.1742-4658.2012.08736.x

  日期：2012.10

  CYP27B1 is a mitochondrial cytochrome P450 that catalyses the hydroxylation of 25‐hydroxyvitamin D3 at the C1α‐position to give the hormonally active form of vitamin D3, 1α,25‐dihydroxyvitamin D3. We successfully expressed human CYP27B1 in Escherichia coli and partially purified this labile enzyme and carried out a detailed characterization of its kinetic properties in a reconstituted membrane environment. The phospholipid concentration did not affect the enzyme activity in the vesicle‐reconstituted system, although it was influenced by the phospholipid composition, with the addition of cardiolipin lowering the Km for 25‐hydroxyvitamin D3. These data are consistent with the enzyme accessing substrate from the hydrophobic domain of the vesicle membrane. Cardiolipin also caused the appearance of inhibition of activity at high substrate concentrations. This substrate inhibition fitted a model for one catalytic and two inhibitory sites on the enzyme for the binding of substrate. The Km for human adrenodoxin was observed to decrease with decreasing substrate concentration, with the catalytic efficiency (kcat/Km) being largely independent of adrenodoxin concentration. Human CYP27B1 was also active on 25‐hydroxyvitamin D2 and on intermediates of the CYP24A1‐mediated inactivation pathway, 24R,25‐dihydroxyvitamin D3, 24‐oxo‐25‐hydroxyvitamin D3 and 24‐oxo‐23,25‐dihydroxyvitamin D3, with all these substrates showing comparable kcat values of 50–71 min−1, similar to 25‐hydroxyvitamin D3. The latter two substrates gave higher Km values than that for 25‐hydroxy‐vitamin D3. The present study shows that human CYP27B1 can be partially purified in an active form with the enzyme displaying high activity towards a range of substrates in a phospholipid vesicle‐reconstituted system that mimics the inner‐mitochondrial membrane.
• 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.