Trifluoroacetic acid → HCID 131732147, compound

HRID 1400966Notes

Reaction equation

HRID 1400966 reaction equation
[CH3:1][C@@H:2]1[O:7][C@@H:6]([O:8][C@@H:9]2[C:14]3=[C:15]([OH:32])[C:16]4[C:28](=[O:29])[C:27]5[C:22](=[CH:23][CH:24]=[CH:25][C:26]=5[O:30][CH3:31])[C:20](=[O:21])[C:17]=4[C:18]([OH:19])=[C:13]3[CH2:12][C@@:11]([OH:37])([C:33]([CH2:35][OH:36])=[O:34])[CH2:10]2)[CH2:5][C@H:4]([NH2:38])[C@@H:3]1[OH:39].C(N([CH2:45][CH3:46])CC)C.C[C@@H]1O[C@@H]([O:54][C@@H:55]2[C:60]3=[C:61]([OH:78])[C:62]4[C:74](=O)[C:73]5C(=CC=C[C:72]=5[O:76]C)C(=O)[C:63]=4C(O)=[C:59]3[CH2:58][C@@:57](O)([C:79](CO)=[O:80])[CH2:56]2)C[C@H](N)[C@@H]1O.Cl.[C:87](O)(C(F)(F)F)=O>CN(C=O)C.C(#N)C.O>[CH3:1][C@@H:2]1[O:7][C@@H:6]([O:8][C@@H:9]2[C:14]3=[C:15]([OH:32])[C:16]4[C:28](=[O:29])[C:27]5[C:22](=[CH:23][CH:24]=[CH:25][C:26]=5[O:30][CH3:31])[C:20](=[O:21])[C:17]=4[C:18]([OH:19])=[C:13]3[CH2:12][C@@:11]([OH:37])([C:33]([CH2:35][OH:36])=[O:34])[CH2:10]2)[CH2:5][C@H:4]([NH2:38])[C@@H:3]1[OH:39].[CH3:87][C@@:23]12[C@H:74]3[CH2:73][C@@H:72]([OH:76])[C@:45]4([CH3:46])[C@@H:58]([C:57]5[CH2:79][O:80][C:55](=[O:54])[CH:56]=5)[CH2:59][CH2:60][C@:61]4([OH:78])[C@@H:62]3[CH2:63][CH2:20][C@@H:22]1[CH2:27][C@@H:26]([OH:30])[CH2:25][CH2:24]2

Participants

7

Reactant 4

HCID 6422Trifluoroacetic acidHCID 8471TriethylamineHCID 31703Doxorubicin2×HCID 443939Doxorubicin Hydrochloride

Product 1

HCID 131732147HCID 131732147, compound

Reagents, catalysts & solvents 2

HCID 6228DimethylformamideSolvent2×HCID 18373917Acetonitrile--water (1/1)Solvent

Procedure

Doxorubicin was obtained from Sigma-Aldrich. To couple digoxygenin to doxorubicin, we performed the following procedure: To a solution of Digoxigenin-3-O-methylcarbonyl-epsilon-aminocaproic acid-N-hydroxysuccinimide ester (20 mg, 30.4 μmol) in DMF (500 μL) was added triethylamine (8.4 μL, 60.8 μmol) and the resulting mixture was transferred immediately to a solution of Doxorubicin-HCl (16.6 mg, 30.4 μmol) in DMF (500 μL). The reaction mixture was tumbled for 2 h at ambient temperature, filtered, and the target compound was isolated by preparative reversed phase HPLC employing an acetonitrile/water gradient containing 0.1% TFA (Merck Cromolith prep RP-18e column, 100×25 mm). After lyophilization the Doxorubicin-Digoxigenin conjugate was obtained as a colorless solid (25.2 mg, 76%). Analytical HPLC: tR=13.9 min (Merck Chromolith Performance RP-18e, 100×4.6 mm, water+0.1% TFA acetonitrile/water+0.1% TFA 80:20, 25 min); ESI-MS (positive ion mode): m/z: calcd for C58H75N2O18: 1087.5; found: 1087.6 [M+H]+. The molecular weight of doxorubicin is 579.98 Da. The molecular weight of the resulting Doxorubicin-Dig conjugate is 1087.24 Da. Until the point of complexation to the antibody, we stored the conjugate in aliquots in DMSO at −20° C. FIG. 18 shows the structure of Doxorubicin-digoxygenin conjugate.

Workup

  1. filtrationfiltered
  2. customthe target compound was isolated by preparative reversed phase HPLC

Sources & evidence

ORD record
ord-8125550e2a1c457bb95462acaaa6b06d
Source dataset
uspto-grants-2014_12
Patent number
US08907069B2

Reaction notes

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