A kind of synthetic method of parathyroid hormone analog
A technology for parathyroid hormone and a synthesis method, which is applied in the field of synthesis of parathyroid hormone analogs, can solve the problems such as the inability to reach a high level of yield and purity, many impurities, and difficulty in purification.
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Embodiment 1
[0069] Example 1: Preparation of Peptide Resin Fragment 1
[0070] 1. Preparation of Fmoc-Thr(tBu)-Rink amide Resin
[0071] Take the Rink amide Resin resin (20g, 12mmol) with a degree of substitution sub=0.6mmol / g, add it to the reaction column, swell it with DMF for more than 30 minutes, drain the DMF, and use DBLK (a DMF solution containing 20% piperidine) Remove Fmoc, each removal reaction is 5 minutes, two times in total, after the removal is completed, wash with DMF 6 times, drain and wait for feeding. Weigh Fmoc-Thr(tBu)-OH (2.4g, 6mmol), HOBt (0.8g, 6mmol) and dissolve it with DMF (30ml) and DCM (30ml) and add DIC (0.9g, 7.2mmol) under ice bath conditions, After activating for 5 minutes, add it to the reaction column. After 60 minutes, the reaction is over. The resin is washed three times with DMF, and the blocking solution (pyridine (20ml) and acetic anhydride (22ml)) is added. After 2 hours of reaction, it is drained and washed with DMF for 6 times. . Then add a...
Embodiment 2
[0074] Embodiment 2: Preparation of peptide fragment 2
[0075] Take by weighing the 2-CTC resin (40g, 20mmol) that substitution degree is 0.5mmol / g, join in the solid-phase reaction column, wash 2 times with DMF, after 30 minutes with DMF swelling resin, take Fmoc-Leu-OH (21.2 g, 60mmol) was dissolved in a mixed solution of DCM (30ml) and DMF (30ml) with a volume ratio of 1:1, DIPEA (15.5g, 120mmol) was added under ice-bath conditions, and the solution was added to the solid-phase reaction column after activation for 5 minutes , reacted at room temperature for 2 hours, and blocked three times with blocking solution (DIPEA:methanol:DCM=1:2:17v:v) after the reaction, 3 minutes each time. The volume of closed liquid is calculated as 4.0ml / gram of resin. Then add an appropriate amount of methanol to wash 3 times, each time for 10 minutes, and wash 6 times with DMF. Fmoc protection was removed with DBLK, followed by 6 washes with DMF.
[0076] Dissolve Fmoc-Glu(OtBu)-OH (25.5g,...
Embodiment 3
[0078] Embodiment 3: the preparation of peptide fragment 3
[0079] Take by weighing the 2-CTC resin (40g, 20mmol) that substitution degree is 0.5mmol / g, join in the solid-phase reaction column, wash 2 times with DMF, after 30 minutes with DMF swelling resin, take Fmoc-Ile-OH (21.2 g, 60mmol) was dissolved in a mixed solution of DCM (30ml) and DMF (30ml) with a volume ratio of 1:1, DIPEA (15.5g, 120mmol) was added under ice-bath conditions, and the solution was added to the solid-phase reaction column after activation for 5 minutes , reacted at room temperature for 2 hours, and blocked three times with blocking solution (DIPEA:methanol:DCM=1:2:17v:v) after the reaction, 3 minutes each time. The volume of closed liquid is calculated as 4.0ml / gram of resin. Then add an appropriate amount of methanol to wash 3 times, each time for 10 minutes, and wash 6 times with DMF. Fmoc protection was removed with DBLK, followed by 6 washes with DMF.
[0080] Dissolve Fmoc-Ser(tBu)-OH (23....
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