Method for preparing etelcalcetide through solid-liquid phase combination
A technology of veracatide and solid-liquid phase, which is applied in the field of solid-liquid phase preparation of veracatide, can solve the problems of unfavorable large-scale industrial production, expensive amino acid materials, high synthesis cost, etc., and achieve the goal of being suitable for large-scale production, The effect of less synthetic impurities and low synthesis cost
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Embodiment 1
[0028] The synthesis of embodiment 1 Fmoc-D-Arg (Pbf)-Rink Amide resin
[0029] Weigh the Rink Amide resin (5.95g, 5mmol) with a substitution degree of 0.84mmol / g, add it to the polypeptide solid-phase reactor, wash it with DMF (30ml) three times, drain it, and add DMF (60ml) to swell for 30min. After filtering, add 20% piperidine / DMF solution to remove the Fmoc protecting group, detect the reaction with ninhydrin, the resin turns blue, the deprotection is complete, drain the solvent, add DMF to wash the resin 6 times.
[0030] Fmoc-D-Arg(Pbf)-OH (8.12g, 12.5mmol) and HOBt (2.54g, 18.75mmol) were weighed and dissolved in 40ml of DMF, DIC (2.86g, 22.50mmol) was added for activation for 5min, and the activated The amino acid solution was added to the above solid-phase reactor, and stirred at room temperature for 2 hours. The reaction was detected with ninhydrin, the resin was colorless, the condensation reaction was complete, the solvent was drained, and DMF was added to wash t...
Embodiment 2
[0031] The synthesis of embodiment 2 Fmoc-D-Arg (Pbf)-Sieber resin
[0032] Sieber resin (7.04 g, 5 mmol) with a substitution degree of 0.71 mmol / g was weighed and added to a polypeptide solid-phase reactor, washed three times with DMF (35 ml), drained, and swelled by adding DMF (75 ml) for 30 min. After filtering, add 20% piperidine / DMF solution to remove the Fmoc protecting group, detect the reaction with ninhydrin, the resin turns blue, the deprotection is complete, drain the solvent, add DMF to wash the resin 6 times.
[0033] Fmoc-D-Arg(Pbf)-OH (8.12g, 12.5mmol) and HOBt (2.54g, 18.75mmol) were weighed and dissolved in 45ml of DMF, DIC (2.86g, 22.50mmol) was added to activate for 5min, and then the activated The amino acid solution was added to the above solid-phase reactor, and stirred at room temperature for 2 hours. The reaction was detected with ninhydrin, the resin was colorless, the condensation reaction was complete, the solvent was drained, and DMF was added to w...
Embodiment 3
[0034] Example 3 Ac-D-Cys(Trt)-D-Ala-D-Arg(Pbf)-D-Arg(Pbf)-D-Arg(Pbf)-D-Ala-D-Arg(Pbf)-Rink Amide Synthesis of resin
[0035] Get the Fmoc-D-Arg(Pbf)-Rink Amide resin obtained in Example 1, according to the coupling method in Example 1, sequentially couple Fmoc-D-Ala-OH, Fmoc-D-Arg(Pbf)-OH, Fmoc-D-Arg(Pbf)-OH, Fmoc-D-Arg(Pbf)-OH, Fmoc-D-Ala-OH and Fmoc-D-Cys(Trt)-OH.
[0036]After the coupling is completed, the Fmoc protecting group is removed. Weigh acetic anhydride (1.55g, 10mmol), add 40ml of DMF and DIEA (1.96g, 10mmol), add the mixed solution to the resin for reaction, detect ninhydrin, the resin is colorless, wash the resin 6 times with DMF. Then wash 3 times with DCM, 40ml / time, after washing, take out the peptide resin and dry it to obtain 15.54g peptide resin.
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