3-Mercaptopropionic Acid → Succinimidyl 4-(N-maleimidomethyl)cyclohexanecarboxylate

HRID 628598

반응식

HRID 628598 반응식
[SH:1][CH2:2][CH2:3][C:4]([OH:6])=[O:5].[CH2:7]1[CH:12]([CH2:13][N:14]2[C:19](=[O:20])[CH:18]=[CH:17][C:15]2=[O:16])[CH2:11][CH2:10][CH:9]([C:21]([O:23][N:24]2[C:29](=[O:30])[CH2:28][CH2:27][C:25]2=[O:26])=[O:22])[CH2:8]1.C(N(CC)C(C)C)(C)C>>[CH2:7]1[CH:12]([CH2:13][N:14]2[C:19](=[O:20])[CH:18]=[CH:17][C:15]2=[O:16])[CH2:11][CH2:10][CH:9]([C:21]([O:23][N:24]2[C:25](=[O:26])[CH2:27][CH2:28][C:29]2=[O:30])=[O:22])[CH2:8]1.[SH:1][CH2:2][CH2:3][C:4]([OH:6])=[O:5].[CH2:7]1[CH:12]([CH2:13][N:14]2[C:19](=[O:20])[CH:18]=[CH:17][C:15]2=[O:16])[CH2:11][CH2:10][CH:9]([C:21]([O:23][N:24]2[C:25](=[O:26])[CH2:27][CH2:28][C:29]2=[O:30])=[O:22])[CH2:8]1

참가 물질

5

반응물 3

HCID 65143-Mercaptopropionic AcidHCID 81531DiisopropylethylamineHCID 125175Succinimidyl 4-(N-maleimidomethyl)cyclohexanecarboxylate

생성물 2

HCID 125175Succinimidyl 4-(N-maleimidomethyl)cyclohexanecarboxylateHCID 131731991MPr SMCC

실험 절차

The drug-linker SMCC-MDC was prepared in the following reactions: (1) 3-mercaptopropanoic acid (MPr) was reacted with N-succinimidyl 4-(maleimidomethyl)cyclohexane-1-carboxylate (SMCC) in the presence of N,N-diisopropylethylamine (DIEA), giving the MPr-SMCC at a yield of over 95%; (2) condensation of N-Me-L-Ala-MDC, which was prepared by deprotection of Fmoc-N-Me-Ala-MDC under a base piperidine in CH3CN, with MPr-SMCC in the presence of a coupling reagent EDC, giving the desired coupled product SMCC-MDC in 60-70% yield over two steps. Anti-EGFR antibody Anti-EGFR antibody was diluted to 2.5 mg/mL in solution A (50 mM potassium phosphate, 50 mM NaCl, and 2 mM EDTA, pH 6.5). SMCC-MDC was added to give a ratio of SMCC-MDC to antibody of 7:1 mole equivalent. Then dimethylacetamide (DMA) was added to 15% (v/v) to the reaction and reaction was mixed by stirring for 4 h at ambient temperature. D-Lmcc-Anti-EGFR antibody conjugate was purified from excess unreacted or hydrolyzed reagent and excess SMCC-MDC using a G25 gel filtration column equilibrated in pH 7.4 phosphate buffer (aqueous). The conjugate was then dialyzed overnight into pH 7.4 phosphate buffer (aqueous) and filtered through a 0.22 μm filter for final storage. The number of SMCC-MDC molecule per antibody molecule in the final conjugate was measured by determining absorbance of the conjugate at 252 and 280 nm and using known extinction coefficients for SMCC-MDC and antibody at these two wavelengths. A ratio of maytansinoid compound to antibody of 3.5:1.0 was normally obtained.

출처 및 근거

ORD 기록
ord-a40b62faaa284d3d93c6f7bae3323b09
출처 데이터셋
uspto-grants-2016_04
특허 번호
US09314536B2

연결된 노트

불러오는 중…