Carbon Dioxide → HCID 11594165, compound

HRID 1176188Notes

Reaction equation

HRID 1176188 reaction equation
CC1[C:11]2([O:19][C:18]3[C:13](=[C:14](O)[CH:15]=[C:16](C=O)[C:17]=3C(O)=O)C2)C2(C)C(C(C)(C)C(O)C(O)C2)CC1.[N:31]1[C:36]2[CH:37]=[CH:38][CH:39]=[CH:40][C:35]=2[C:34]([NH:41][CH2:42]C2C=CC(OC)=CC=2)=[N:33][N:32]=1.C(=O)=O>CS(C)=O>[N:31]1[C:36]2[CH:37]=[CH:38][CH:39]=[CH:40][C:35]=2[C:34]([N:41]([C:15]2[CH:14]=[CH:13][C:18]([O:19][CH3:11])=[CH:17][CH:16]=2)[CH3:42])=[N:33][N:32]=1

Participants

5

Reactant 3

HCID 280Carbon DioxideHCID 1195286-Formyl-4,6',7'-trihydroxy-2',5',5',8'a-tetramethyl-3',4',4'a,5',6',7',8',8'a-octahydro-2'H,3H-spiro[1-benzofuran-2,1'-naphthalene]-7-carboxylic acidHCID 578097HCID 578097, compound

Product 1

HCID 11594165HCID 11594165, compound

Reagents, catalysts & solvents 1

HCID 679Dimethyl SulfoxideSolvent2×

Procedure

T-47D, HT29, H1299, MX-1 and MDAMB435 cells were grown and harvested as in Example 29. An aliquot of 90 μL of cells (4.4×104 cells/mL) was added to a well of a 96-well microtiter plate containing 5 μL of a 10% DMSO in RPMI-1640 media solution containing 10 nM to 100 μM of benzo[d][1,2,3]triazin-4-yl-(4-methoxy-phenyl)methyl-amine (1 nM to 10 μM final). An aliquot of 45 μL of cells was added to a well of a 96-well microtiter plate containing 5 μL of a 10% DMSO in RPMI-1640 media solution without compound as the control sample for maximal cell proliferation (LMax). The samples were mixed by agitation and then incubated at 37° C. for 48 h in a 5% CO2-95% humidity incubator. After incubation, the samples were removed from the incubator and 25 μL of CellTiter-Glo™ reagent (Promega) was added. The samples were mixed by agitation and incubated at room temperature for 10-15 min. Plates were then read using a luminescent plate reader (Model SPECTRAfluor Plus, Tecan) to give Ltest values.

Workup

  1. additionAn aliquot of 90 μL of cells (4.4×104 cells/mL) was added to a well of a 96-well microtiter plate
  2. additionAn aliquot of 45 μL of cells was added to a well of a 96-well microtiter plate
  3. additionThe samples were mixed by agitation
  4. customAfter incubation, the samples were removed from the incubator and 25 μL of CellTiter-Glo™ reagent (Promega)
  5. additionwas added
  6. additionThe samples were mixed by agitation
  7. customto give Ltest values

Sources & evidence

ORD record
ord-9e0bbd7960d94ef7b60f6a3316a4d7ea
Source dataset
uspto-grants-2010_11
Patent number
US07842805B2

Reaction notes

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