Phosphate → HCID 92131096, compound

HRID 1748362Notes

Reaction equation

HRID 1748362 reaction equation
[CH3:1][C@H:2]1[C@@H:27]2[O:28][C@@:26]2([CH3:29])[C@@H:25]([O:30][C:31]([C@@H:33]([N:35]([C:37]([CH2:39][CH2:40][SH:41])=[O:38])[CH3:36])[CH3:34])=[O:32])[CH2:24][C:22](=[O:23])[N:21]([CH3:42])[C:14]2=[C:15]([Cl:20])[C:16]([O:18][CH3:19])=[CH:17][C:12](=[CH:13]2)[CH2:11][C:10]([CH3:43])=[CH:9][CH:8]=[CH:7][C@@H:6]([O:44][CH3:45])[C@:5]2([OH:50])[NH:46][C:47]([O:49][C@H:3]1[CH2:4]2)=[O:48].[CH2:51]1[CH:56]([CH2:57][N:58]2[C:63](=[O:64])[CH:62]=[CH:61][C:59]2=[O:60])[CH2:55][CH2:54][CH:53]([C:65]([O:67][N:68]2[C:73](=[O:74])[CH2:72][CH2:71][C:69]2=[O:70])=[O:66])[CH2:52]1.P([O-])([O-])([O-])=O.P([O-])([O-])([O-])=O.[K+].[K+].[K+].C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O>C1COCC1>[CH3:1][C@H:2]1[C@@H:27]2[O:28][C@@:26]2([CH3:29])[C@@H:25]([O:30][C:31]([C@@H:33]([N:35]([C:37]([CH2:39][CH2:40][S:41][CH:62]2[C:63](=[O:64])[N:58]([CH2:57][CH:56]3[CH2:55][CH2:54][CH:53]([C:65]([O:67][N:68]4[C:73](=[O:74])[CH2:72][CH2:71][C:69]4=[O:70])=[O:66])[CH2:52][CH2:51]3)[C:59](=[O:60])[CH2:61]2)=[O:38])[CH3:36])[CH3:34])=[O:32])[CH2:24][C:22](=[O:23])[N:21]([CH3:42])[C:14]2=[C:15]([Cl:20])[C:16]([O:18][CH3:19])=[CH:17][C:12](=[CH:13]2)[CH2:11][C:10]([CH3:43])=[CH:9][CH:8]=[CH:7][C@@H:6]([O:44][CH3:45])[C@:5]2([OH:50])[NH:46][C:47]([O:49][C@H:3]1[CH2:4]2)=[O:48]

Participants

7

Reactant 5

HCID 1061PhosphateHCID 6049Ethylenediaminetetraacetic AcidHCID 62657Potassium Phosphate, TribasicHCID 125175Succinimidyl 4-(N-maleimidomethyl)cyclohexanecarboxylateHCID 11343137Mertansine

Product 1

HCID 92131096HCID 92131096, compoundYield 63.9%

Reagents, catalysts & solvents 1

HCID 8028TetrahydrofuranSolvent

Procedure

A round bottom flask was charged with N2′-Deacetyl-N2′-(3-mercapto-1-oxopropyl)-maytansine (DM1, 67.9 mg, 0.092 mmol), Succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC, 32.1 mg, 0.096 mmol, 1.05 equivalents) and THF (4 mL). The solution stirred as the reagents were dissolved in the solvent to yield a clear, colorless solution. Phosphate buffer, pH 6, (4 mL, 100 mM Potassium Phosphate, 2 mM EDTA) was then added. The reaction flask was equipped with a septum and stir bar and the reaction proceeded at room temperature with vigorous stirring. The reaction appeared complete within 30 minutes as indicated by reverse-phase HPLC. Following reaction completion, the reaction volume was reduced in vacuo to give a white solid/residue. The product was isolated by silica gel chromatography, eluting with a mixture of 3% methanol in methylene chloride. Product containing fractions were combined and concentrated to dryness in vacuo to give 63 mg (63.9% yield) DM1-SMCC as a white solid. Mass spectral analysis of the isolated product gave the expected m+Na+ (m/z 1094.4) and m+Cl− (m/z 1106.2) major molecular ions (FIG. 43).

Workup

  1. dissolutionwere dissolved in the solvent
  2. customto yield a clear, colorless solution
  3. customThe reaction flask was equipped with a septum
  4. stirringstir bar
  5. customproceeded at room temperature with vigorous stirring
  6. customreaction completion
  7. customto give a white solid/residue
  8. customThe product was isolated by silica gel chromatography
  9. washeluting with a mixture of 3% methanol in methylene chloride
  10. additionProduct containing fractions
  11. concentrationconcentrated to dryness in vacuo

Sources & evidence

ORD record
ord-8dc302663a3840578be694c1f4ce24a1
Source dataset
uspto-grants-2015_10
Patent number
US09150649B2

Reaction notes

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