Carbon Dioxide → Prostaglandin E2

HRID 2069545Notes

Reaction equation

HRID 2069545 reaction equation
C1C=C2C(C3C=CC(O)=CC=3)(C3C=CC(O)=CC=3)OS(=O)(=O)C2=CC=1.N[C@H:27]([C:33]([OH:35])=[O:34])[CH2:28][CH2:29][C:30](=O)N.C(=O)=O.C[C@H:40]1[C@@H:61]([CH2:62]C2OC3C=CC(NC)=C(C(O)=O)C=3N=2)[O:60][C@:43]2([O:48][C@H:47]([C@@H:49]([C:51]([C:53]3NC=[CH:55][CH:54]=3)=[O:52])C)[C@H:46](C)[CH2:45][C@H:44]2C)[CH2:42][CH2:41]1>CS(C)=O>[CH3:62][CH2:61][CH2:40][CH2:41][CH2:42][C@H:43]([OH:60])/[CH:44]=[CH:45]/[C@@H:46]1[C@@H:53]([CH2:54]/[CH:55]=[CH:30]\[CH2:29][CH2:28][CH2:27][C:33]([OH:35])=[O:34])[C:51](=[O:52])[CH2:49][C@H:47]1[OH:48]

Participants

6

Reactant 4

HCID 280Carbon DioxideHCID 4766Phenol RedHCID 5961L-GlutamineHCID 11957499Calcimycin

Product 1

HCID 5280360Prostaglandin E2

Reagents, catalysts & solvents 1

HCID 679Dimethyl SulfoxideSolvent

Procedure

3T3 Swiss Albino fibroblast cell suspension (Phenol-red free DMEM with 10% FBS and 2 mM L-glutamine) was plated out into 96-well tissue culture plates (1×105 cells/mL, 100 μL/well). The cells were cultured overnight at 37° C., 5% CO2. The extracts and fractions were solubilised in DMSO, and diluted appropriately with media so the extracts were tested at a final concentration of 1000, 10, and 1 μg/mL; 5 μL of sample was added to each well. Cells+samples were incubated (37° C., 3 hours), before the addition of calcium ionophore A23187. Following 20 min incubation, the plate was centrifuged (1000 g, 3 min) and the supernatants were removed. Included on the plate was a positive control (aspirin; 100 μM), and DMSO control, both with and without calcium ionophore A23187.

Workup

  1. additiondiluted appropriately with media so the extracts
  2. addition5 μL of sample was added to each well
  3. custom(37° C., 3 hours)
  4. waitFollowing 20 min
  5. custom(1000 g, 3 min)
  6. customthe supernatants were removed

Sources & evidence

ORD record
ord-488bcfd35b504fb5aa8dff68f4f9c95f
Source dataset
uspto-grants-2012_10
Patent number
US08299119B2

Reaction notes

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