Ethylenediaminetetraacetic Acid → Succinimidyl 4-(N-maleimidomethyl)cyclohexanecarboxylate

HRID 1482731

Reaction equation

HRID 1482731 reaction equation
[CH2:1]1[CH:6]([CH2:7][N:8]2[C:13](=[O:14])[CH:12]=[CH:11][C:9]2=[O:10])[CH2:5][CH2:4][CH:3]([C:15]([O:17][N:18]2[C:23](=[O:24])[CH:22](S(O)(=O)=O)[CH2:21][C:19]2=[O:20])=[O:16])[CH2:2]1.C1C(CN2C(=O)C=CC2=O)CCC(C(ON2C(=O)C(S([O-])(=O)=O)CC2=O)=O)C1.[Na+].C([O-])(=O)CCC([O-])=O.C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O>CC(N(C)C)=O.C1N(CCCS(O)(=O)=O)CCN(CCO)C1>[CH2:1]1[CH:6]([CH2:7][N:8]2[C:13](=[O:14])[CH:12]=[CH:11][C:9]2=[O:10])[CH2:5][CH2:4][CH:3]([C:15]([O:17][N:18]2[C:19](=[O:20])[CH2:21][CH2:22][C:23]2=[O:24])=[O:16])[CH2:2]1

Participants

7

Reactant 4

HCID 6049Ethylenediaminetetraacetic AcidHCID 160419SuccinateHCID 4618794Sulfo-SMCC sodiumHCID 46187951-[4-[(2,5-Dioxopyrrol-1-yl)methyl]cyclohexanecarbonyl]oxy-2,5-dioxopyrrolidine-3-sulfonic acid2×

Product 1

HCID 125175Succinimidyl 4-(N-maleimidomethyl)cyclohexanecarboxylate

Reagents, catalysts & solvents 2

HCID 31374DimethylacetamideSolvent2×HCID 85255HeppsSolvent

Procedure

The conjugates of the present invention can also be prepared by in situ process according to the following procedures. Antibodies (21 clones) were conjugated to DM1 using the sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC) linker Stock solutions of DM1 and sulfo-SMCC heterobifunctional linker were prepared in DMA. Sulfo-SMCC and DM1 thiol were mixed together to react for 10 minutes at 25° C. in DMA containing 40% v/v of aqueous 50 mM succinate buffer, 2 mM EDTA, pH 5.0, at the ratio of DM1 to linker of 1.3:1 mole equivalent and a final concentration of DM1 of 1.95 mM. The antibody was then reacted with an aliquot of the reaction to give a mole equivalent ratio of SMCC to Ab of around 6.5:1 under final conjugation conditions of 2.5 mg/mL of Ab in 50 mM EPPS, pH 8.0 and 10% DMA (v/v). After approximately 18 hours at 25° C., the conjugation reaction mixture was purified using a SEPHADEX™ G25 column equilibrated with 10 mM succinate, 250 mM glycine, 0.5% sucrose, 0.01% Tween 20, pH 5.5.

Workup

  1. customThe conjugates of the present invention can also be prepared by in situ process
  2. customThe antibody was then reacted with an aliquot of the reaction

Sources & evidence

ORD record
ord-0646637369f04b9a99367913c136d306
Source dataset
uspto-grants-2016_08
Patent number
US09415118B2

Linked notes

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