Enzymatic type cleavable PEGylation ( polyethylene glycol) lipid material
A lipid material and enzymatic technology, applied in the direction of liposome delivery, genetic material components, non-active components of polymer compounds, etc.
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Embodiment 1
[0051] Synthesis of Cholesterol Bromoacetyl Ester (1)
[0052] 0.5 mmol of cholesterol (193 mg) was dissolved in 5 ml of anhydrous THF, and 3 mmol of bromoacetyl chloride (250 μl) was added dropwise. The reaction mixture was heated, stirred and refluxed for about 4h, and detected by TLC (petroleum ether: CH 2 Cl 2 =4:1) The reaction is complete. The organic solvent was evaporated by rotary evaporation to obtain a yellow crude product, which was then purified by a silica gel column (eluent was sherwood oil: CH 2 Cl 2 =5:1). Finally, a white sample is obtained, that is, Chol-CO-CH 2 Br.
[0053] 1H NMR (CDCl3, 400 MHz): δ0.6 —2.25ppm(m, 43H, cholesterol protons), 3.8ppm(s, 2H, CH2Br), 4.6ppm(m, 1H, 3-CH-cholesterol), 5.4ppm (d, 1H, 6-CH-cholesterol).
Embodiment 2
[0055] Synthesis of Cholesterol Bromoacetyl Ester (2)
[0056] Dissolve 100 mg cholesterol and 40 mg bromoacetic acid in 10 ml anhydrous CH 2 Cl 2 , and then added 44 μl of N,N'-diisopropylcarbimide and 1.5 mg of 4-dimethylaminopyridine. Stir at room temperature for 48h. The organic solvent was evaporated by rotary evaporation to obtain a yellow crude product, which was then purified by a silica gel column (eluent was sherwood oil: CH 2 Cl 2 =5:1). Finally, a white solid is obtained, namely Chol-CO-CH 2 Br.
[0057] 1 H NMR (CDCl 3 , 400 MHz): δ0.6 —2.25ppm(m, 43H, cholesterol protons), 3.8ppm(s, 2H, CH2Br), 4.6ppm(m, 1H, 3-CH-cholesterol), 5.4ppm(d, 1H , 6-CH-cholesterol).
Embodiment 3
[0059] Synthesis of PEG-peptide-SH (Ⅰ)
[0060] mPEG2000-NHS (purchased from Sigma, USA) and short peptide (GPLGIAGQC)-SH (synthesized by Shanghai Chueptide Biotechnology Co., Ltd.) were mixed at a molar ratio of 1.2:1, stirred in carbonic acid buffer (pH 8.5) at 4°C overnight. After dialysis (dialysis bag molecular weight cut-off 1000) and freeze-drying, a white solid, PEG2000-peptide(GPLGIAGQC)-SH (I), was obtained. ( figure 2 PEG-MMP substrate peptide 1 H NMR spectrum)
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