Double-chain structured polyethylene active derivatives, and ligature with other molecules
A polyethylene glycol and derivative technology, applied in dipeptide components, cyclic peptide components, peptide/protein components, etc., can solve the problems of linking group hydrolysis, fragmentation, protein inactivation, etc.
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Embodiment 1
[0027] Example 1: Preparation of N, N-bispolyethylene glycol acetamido-butyric acid succinimidyl ester 1000
[0028] 2g N,N-dipolyethylene glycol acetamide 1000 (0.002mol) and 0.53g 4-chlorobutyric acid (0.005mol) were dissolved in 50ml of dichloromethane, refluxed at room temperature for 2 hours, and the mixture was concentrated under reduced pressure to 20ml, add 200ml of cooling diethyl ether, filter the resulting precipitate and dry under reduced pressure to obtain 1.75g of the product N,N-bispolyethylene glycol acetamidobutyric acid 1000.
[0029] Get above-mentioned obtained product and 0.58g N-hydroxysuccinimide (0.005mol) and dissolve in 50ml dichloromethane, in the presence of dimethylaminopyridine (DMAP) and dicyclohexylcarbodiimide (DCC), temperature- 20~10℃, react for 4 hours or overnight. The mixture was precipitated with ether, recrystallized from ethanol, and further purified through a Sephadex G-25 column to obtain 1.7 g of pure N,N-bispolyethylene glycol ac...
Embodiment 2
[0030] Embodiment 2: the preparation of N, N-dipolyethylene glycol acetamido-butyric acid maleimide ester 20000
[0031]2g N, N-dipolyethylene glycol acetamide 1000 (0.002mol) and 0.53g 1-methyl-4-chlorobutyric acid (0.005mol) were dissolved in 50ml of dichloromethane, refluxed at room temperature for 2 hours, and The mixture was concentrated under reduced pressure to 20 ml, 200 ml of diethyl ether was added for cooling, and the obtained precipitate was filtered and then dried under reduced pressure to obtain 1.75 g of the product N,N-bispolyethylene glycol acetamidomethyl butyric acid 1000. The above product and 0.57g N-hydroxyl maleimide (0.005mol) were dissolved in 50ml of methylene chloride, in the presence of dimethylaminopyridine (DMAP) and dicyclohexylcarbodiimide (DCC), the temperature- 20~10℃, react for 4 hours or overnight. The mixture was precipitated with ether, recrystallized from ethanol, and further purified by Sephadex G-25 column to obtain pure N,N-bispolyeth...
Embodiment 3
[0032] Example 3: Preparation of N,N-bispolyethylene glycol acetamidoacetic acid-6-aminocaproic acid succinimidyl ester 60000
[0033] Get 60g N, N-bispolyethylene glycol acetamidoacetic acid 40000 (0.001mol), 0.65g aminon-hexanoic acid (0.005mol) and 1.03g dicyclohexylcarbodiimide (DCC, 0.005mol) are dissolved in 100ml of dichloromethane, stirred and refluxed at room temperature for 16 hours. The mixture was filtered, the obtained filtrate was concentrated to 20ml under reduced pressure, 200ml of cooling diethyl ether was added, the obtained precipitate was filtered and then dried under reduced pressure to obtain the product N,N-dipolyethylene glycol acetamidoacetic acid-6-aminocaproic acid 40000 52.5 g.
[0034] Get above-mentioned gained product and 0.58g N-hydroxysuccinimide (0.005mol) and dissolve in 50ml methylene chloride, in the presence of dimethylaminopyridine (DMAP) and dicyclohexylcarbodiimide (DCC), Temperature -20~10°C, react for 4 hours or overnight. The mixt...
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