Method for synthesizing Etelcalcetide
A d-cys and fragment technology, applied in the field of synthesis of Etelcalcetide, can solve the problems of low total yield, high production cost, cumbersome reaction steps, etc., and achieve the improvement of total yield, high purity and yield, crude peptide yield and The effect of high purity
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Embodiment 1
[0043] Example 1: N-Ac-D-Cys(Mmt)-D-Ala-D-Arg(Pbf)-D-Arg(Pbf)-D-Arg(Pbf)-D-Ala-D-Arg(Pbf) -Synthesis of Rink-Amide Resin
[0044] Weigh 25.00 g of Rink-Amide resin with a substitution degree of 0.20 mmol / g, add it to a solid-phase reaction column, wash it twice with DMF, swell the resin with DMF for 30 minutes, add 20% piperidine / DMF (V / V) Remove Fmoc from the solution for 5+10 minutes, wash the resin 6 times with DMF after removal, and detect the color of the resin with ninhydrin. Weigh 16.30g (25mmol) Fmoc-D-Arg(Pbf)-OH, 4.05g (30mmol) HOBt, 9.51g (25mmol) HBTU, dissolve with 75ml DMF and 75ml DCM, add 6.5ml (37.5mmol) under ice water bath DIPEA was activated for 5 minutes, and the mixture was added to the reaction column, and reacted at room temperature for 2 hours, and the end of the reaction was detected with ninhydrin (if the resin is colorless and transparent, stop the reaction; if the resin develops color, prolong the reaction for 1 hour, the same below).
[0045] Af...
Embodiment 2
[0047] Example 2: N-Ac-D-Cys(Mmt)-D-Ala-D-Arg(Pbf)-D-Arg(Pbf)-D-Arg(Pbf)-D-Ala-D-Arg(Pbf) -Synthesis of Rink-Amide Resin
[0048] Weigh 11.10 grams of Rink-Amide resin with a substitution degree of 0.45 mmol / g, add it to a solid-phase reaction column, wash it twice with DMF, swell the resin with DMF for 30 minutes, add 20% piperidine / DMF (V / V) Remove Fmoc from the solution for 5+10 minutes, wash the resin 6 times with DMF after removal, and detect the color of the resin with ninhydrin. Weigh 16.30g (25mmol) Fmoc-D-Arg(Pbf)-OH, 4.05g (30mmol) HOBt, 9.51g (25mmol) HBTU, dissolve with 75ml DMF and 75ml DCM, add 6.5ml (37.5mmol) under ice water bath DIPEA was activated for 5 minutes, the mixture was added to the reaction column, and reacted at room temperature for 2 hours, and the end of the reaction was detected by ninhydrin.
[0049] After the reaction is finished, wash the resin 3 times with DMF, add 20% piperidine / DMF (V / V) solution for 5+10 minutes to remove Fmoc, wash the ...
Embodiment 3
[0051] Example 3: N-Ac-D-Cys(Mmt)-D-Ala-D-Arg(Pbf)-D-Arg(Pbf)-D-Arg(Pbf)-D-Ala-D-Arg(Pbf) -Synthesis of Rink-Amide Resin
[0052] Weigh 8.33 grams of Rink-Amide resin with a substitution degree of 0.60 mmol / g, add it to the solid phase reaction column, wash it twice with DMF, swell the resin with DMF for 30 minutes, add 20% piperidine / DMF (V / V) Remove Fmoc from the solution for 5+10 minutes, wash the resin 6 times with DMF after removal, and detect the color of the resin with ninhydrin. Weigh 16.30g (25mmol) Fmoc-D-Arg(Pbf)-OH, 4.05g (30mmol) HOBt, 9.51g (25mmol) HBTU, dissolve with 75ml DMF and 75ml DCM, add 6.5ml (37.5mmol) under ice water bath DIPEA was activated for 5 minutes, the mixture was added to the reaction column, and reacted at room temperature for 2 hours, and the end of the reaction was detected by ninhydrin.
[0053] After the reaction is finished, wash the resin 3 times with DMF, add 20% piperidine / DMF (V / V) solution for 5+10 minutes to remove Fmoc, wash the...
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