Dimethyl Sulfoxide → Corticosterone

HRID 918806Notes

Reaction equation

HRID 918806 reaction equation
CS(C)=[O:3].[CH3:5][C@@:6]12[C@@H:14]([C:15]([CH2:17][OH:18])=[O:16])[CH2:13][CH2:12][C@H:11]1[C@@H:10]1[CH2:19][CH2:20][C:21]3[C@@:27]([CH3:28])([C@H:9]1[CH2:8][CH2:7]2)[CH2:26][CH2:25][C:23](=[O:24])[CH:22]=3>C(O)C>[CH3:28][C@@:27]12[C@H:9]3[C@@H:8]([OH:3])[CH2:7][C@:6]4([CH3:5])[C@@H:14]([C:15]([CH2:17][OH:18])=[O:16])[CH2:13][CH2:12][C@H:11]4[C@@H:10]3[CH2:19][CH2:20][C:21]1=[CH:22][C:23](=[O:24])[CH2:25][CH2:26]2

Participants

4

Reactant 2

HCID 679Dimethyl SulfoxideHCID 6166Deoxycorticosterone

Product 1

HCID 5753Corticosterone

Reagents, catalysts & solvents 1

HCID 702EthanolSolvent2×

Conditions

Temperature
32 °C

Procedure

A suspension of fission yeast (S. pombe PE1) with a cell density of 3·107 cells/ml was prepared on a freshly grown culture using fresh EMMG (pH 7.4) as modified according to Ehmer et al. (Ehmer, P. B. et al., 1. Steroid. Biochem. Mol. Biol. 81, 173-179 (2002)). 492.5 μl of this cell suspension was admixed with 5 μl of inhibitor solution (50 μM of the compound to be tested in ethanol or DMSO) and incubated at 32° C. for 15 min. Controls were admixed with 5 μl of ethanol. The enzyme reaction was started by adding 2.5 μl of 11-deoxycorticosterone (20 μM, containing 1.25 nCi of [4-14C]11-deoxycorticosterone in Ethanol), followed by horizontal shaking at 32° C. for 6 h. The test was quenched by extracting the sample with 500 μl of EtOAc. After centrifugation (10,000 g, 2 min), the EtOAc phase was removed and evaporated to dryness. The residue was taken up in 10 μl of chloroform. The reaction of the substrate to form corticosterone was analyzed by HPTLC (see below).

Workup

  1. additionA suspension of fission yeast (S. pombe PE1) with a cell density of 3·107 cells/ml
  2. customwas prepared on a freshly grown culture
  3. customThe enzyme reaction
  4. customThe test was quenched
  5. extractionby extracting the sample with 500 μl of EtOAc
  6. customAfter centrifugation (10,000 g, 2 min), the EtOAc phase was removed
  7. customevaporated to dryness
  8. customThe reaction of the substrate

Sources & evidence

ORD record
ord-f919ac1c64034673b0bb16bc29acf326
Source dataset
uspto-grants-2016_03
Patent number
US09271963B2

Reaction notes

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