Solid phase synthesis method for thymosin beta 4
A solid-phase synthesis, thymosin technology, applied in the field of biochemistry, can solve the problems of difficulty in purification, affecting the condensation rate of amino acids, increasing the probability of amino acid racemization, etc., achieving the effects of less impurities, shortening synthesis time, and easy separation and purification.
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[0041] Example 1: Preparation of Fmoc-Ser(tBu)-king resin
[0042] Operation: (1) 12.5g (5mmol) of Wang resin (Tianjin Nankai Hecheng Technology Co., Ltd.) was placed in a peptide synthesis reactor, washed with N,N-dimethylformamide (DMF), and then added DMF to swell for 30 minutes After extraction, 3.8g of Fmoc-Ser(tBu)-OH was dissolved in 50ml DMF and added to the reactor, 3.2ml pyridine was added, and 2.8ml 2,6-dichlorobenzoyl chloride was added dropwise. After the addition, the reaction was carried out at room temperature After 2 hours, it was washed three times with DMF, once with methanol, three times with dichloromethane, three times with methanol, and drained to obtain 14.3 g of Fmoc-Ser(tBu)-king resin.
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[0043] Example 2: Fmoc-Thr(tBu)-Ile-Glu(OtBu)-Gln(Trt)-Glu(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gly-Glu(OtBu)-Ser(tBu )-Wang resin (A peptide fully protected resin) preparation
[0044] Operation: (1) 14.3g of Fmoc-Ser(tBu)-king resin was placed in a peptide synthesis reactor, washed with dichloromethane and methanol each 80ml alternately twice, 2 minutes per time, and drained. Add 80ml of 20% piperidine / DMF solution, stir and react at room temperature for 20 minutes, and drain. The resin was washed alternately with dichloromethane and methanol 80ml 3 times, 2 minutes per time, and drained.
[0045] (2) Use Fmoc protected amino acid and O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroborate (TBTU, Shanghai Yanchang Biochemical Technology Development Co., Ltd.) 8.0g in 50ml After the DMF solution is dissolved, add 4.4ml of N,N-diisopropylethylamine, mix well, add to the resin, and stir at room temperature for 3 hours.
[0046] (3) Drain dry. The resin was washed alternately with dichlo...
Example Embodiment
[0051] Example 3: Preparation of Fmoc-Glu(OtBu)-2-chlorotrityl chloride resin
[0052]Operation: Add 100g (120mmol) of 2-chlorotrityl chloride resin to a 1000ml solid phase synthesis reactor. After dissolving 204g of Fmoc-Glu(OtBu)-OH with 600ml of dichloromethane, add N,N-diisopropylethylamine ( DIEA) 167ml, stir and mix, add to the solid phase synthesis reactor, react at room temperature for 1 hour. Drained. Wash the resin alternately with dichloromethane and 800ml methanol for 3 times, 2 minutes / time, and drain. 143.7 g of Fmoc-Glu(OtBu)-2-chlorotrityl chloride resin was obtained.
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