反応式
[CH3:1][C:2]([NH:4][C@H:5]1[C@H:10]([C@H:11]([OH:16])[C@H:12]([OH:15])[CH2:13][OH:14])[O:9][C@:8]([O:20]P(OC[C@H]2O[C@@H](N3C(=O)N=C(N)C=C3)[C@H](O)[C@@H]2O)(O)=O)([C:17]([OH:19])=[O:18])[CH2:7][C@@H:6]1[OH:41])=[O:3].OC(C1(O[C@@H]([C@@H]([C@@H](CO)O)O)[C@H](NC(C)=O)[C@@H](O)C1)O)=O.[O:63]([C@@H]1[C@@H](CO)O[C@@H](O)[C@H](NC(C)=O)[C@H]1O)[C@@H:64]1[O:72][C@H:71]([CH2:73][OH:74])[C@H:69]([OH:70])[C@H:67](O)[C@H:65]1[OH:66].O=C[C@@H]([C@H]([C@H]([C@@H](CO)O)O)O)O>>[OH:19][C:17]([C@@:8]1([O:9][C@@H:10]([C@@H:11]([C@@H:12]([CH2:13][OH:14])[OH:15])[OH:16])[C@H:5]([NH:4][C:2]([CH3:1])=[O:3])[C@@H:6]([OH:41])[CH2:7]1)[O:20][C@H:67]1[C@@H:69]([OH:70])[C@@H:71]([CH2:73][OH:74])[O:72][C@@H:64]([OH:63])[C@@H:65]1[OH:66])=[O:18]参加物
実験手順
The resulting CMP-sialic acid analogue, illustrated in FIG. 28 as the CMP derivative of Neu5Ac (i.e., CMP-Neu5Ac), is then combined with the derivatized βGal(1→4)βGlcNAc-OR compound in the presence of the βGal(1→3/4)βGlcNAc α(2→3)sialyltransferase under conditions wherein sialic acid is transferred to the 3 position of the galactose to form a αNeu5Ac(2-3)βGal- linkage. Suitable conditions, known in the art, include the addition of the sialyltransferase to a mixture of the derivatized βGal(1→4)βGlcNAc-OR compound and of the CMP-sialic acid in a appropriate buffer such as 0.1M sodium cacodylate in appropriate conditions of pH and temperature such as at a pH of 6.5 to 7.5 and a temperature between 25° and 45° C., preferably 35°-40° C., while incubating for 12 hours to 4 days. The resulting sialylated product can be isolated and purified using conventional methodology comprising HPLC, ion exchange-, gel-, reverse-phase-or adsorption chromatography.
出典と根拠
- ORDレコード
- ord-076e515f5b624699a9cadfa0750b2456
- 出典データセット
- uspto-grants-1997_07
- 特許番号
- US05646123
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