A kind of solid-liquid synthesis method of buserelin
A technology of solid-liquid synthesis and solid-phase synthesis, which is applied in the field of solid-liquid synthesis of buserelin, can solve the problems of easy introduction of heavy metals into products, unfavorable industrial production, and high operation requirements, so as to reduce special equipment requirements, improve synthesis efficiency, The effect of safe and easy operation
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Embodiment 1
[0030] Example 1: Preparation of Fmoc-Tyr(tBu)-dichlorotrityl chloride resin.
[0031]Weigh 0.5g (0.30mmol) of dichlorotrityl chloride resin (substitution value 0.60mmol / g) into a 20ml BD syringe (or common glass peptide reactor) with a sieve plate, and swell with 3 times the volume of the resin in DCM Twice, 1 hour each time, to make the resin swell completely. Dissolve 551.40mg (1.2mmol) Fmoc-Tyr(tBu)-OH in 10ml DMF, add 0.4ml (2.4mmol) DIEA, activate for 3min, add the mixed solution into the solid phase reactor, stir for 2h, wash with DMF three times, Wash three times with DCM. Add 10 ml of DCM and 800 μl of methanol-blocked resin, stir for 30 min, wash with DCM for 3 times, and drain to obtain Fmoc-Tyr(tBu)-dichlorotrityl chloride resin.
Embodiment 2
[0032] Example 2: Synthesis of Pyr-His(Trt)-Trp(Boc)-Ser(tBu)-Tyr(tBu)-dichlorotrityl chloride resin.
[0033] Fmoc-Tyr (tBu)-dichlorotrityl chloride resin is loaded into a solid-phase reactor, washed twice with DMF, and the Fmoc protecting group is removed twice with 20% (v / v) piperidine / DMF solution, and the time is 5min and 20min respectively, washed 3 times with DMF, and tested by ninhydrin method. Dissolve 460.08mg (1.2mmol) Fmoc-Ser(tBu)-OH and 162.12mg (1.2mmol) HOBt in 10ml DMF, ice bath for 10min, add 0.4ml (2.4mmol) DIEA, activate for 3min, then add 455.04mg (1.2mmol) ) HBTU, the mixed solution was added to the solid-phase reactor, stirred for 1.5h, and the ninhydrin test was negative, washed three times with DMF and three times with DCM.
[0034] Repeat the above operation, connect Fmoc-Trp(Boc)-OH, Fmoc-His(Trt)-OH, H-Pyr-OH in sequence, the amount of raw materials: amino acid (1.2mmol), HBTU 455.04mg (1.2mmol), HOBt 162.12mg (1.2 mmol). Example 3: Synthesis of ...
Embodiment 3
[0034] Repeat the above operation, connect Fmoc-Trp(Boc)-OH, Fmoc-His(Trt)-OH, H-Pyr-OH in sequence, the amount of raw materials: amino acid (1.2mmol), HBTU 455.04mg (1.2mmol), HOBt 162.12mg (1.2 mmol). Example 3: Synthesis of Fmoc-D-Ser(tBu)-Leu-Arg(HCl)-dichlorotrityl chloride resin.
[0035] Weigh 0.5g (0.30mmol) of dichlorotrityl chloride resin (substitution value 0.60mmol / g) into a 20ml BD syringe (or common glass peptide reactor) with a sieve plate, and swell with 3 times the volume of the resin in DCM Twice, 1 hour each time, to make the resin swell completely. Dissolve 519.48mg (1.2mmol) Fmoc-Arg(HCl)-OH in 10ml DMF, add 0.4ml (2.4mmol) DIEA, activate for 3min, add the mixed solution into the solid phase reactor, stir for 2h, wash with DMF three times, Wash three times with DCM. Add 10 ml of DCM and 800 μl of methanol-blocked resin, stir for 30 min, wash with DCM for 3 times, and drain to obtain Fmoc-Arg(HCl)-dichlorotrityl chloride resin.
[0036] The Fmoc-Arg (HC...
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