The synthetic method of bremelanotide acetate
A technology for bremerondan acetate and a synthesis method, which is applied in the field of polypeptide preparation, can solve the problems of large steric hindrance of the peptide chain, difficulty in forming a ring, and restrict the application of bremerondan, and achieves improvement of purification yield and purity, and chromatographic performance. Good peak shape symmetry and improved separation selectivity
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Embodiment 1
[0076] A preparation method of RinkAmide resin modified by 4-trifluoromethylmandelic acid:
[0077] 1) Rinse 100 g of RinkAmide resin (with a substitution degree of 0.45 mmol / g and a cross-linking degree of 1%) with DCM for 5 times, soak in 500 mL of DCM for 20 min to fully swell, and add the resin to 500 mL of piperidine / DCM mixed solution (the mass percentage of piperidine is 25%), stirred and reacted for 30min to de-Fmoc protect the resin;
[0078] 2) Add 22 g of 4-trifluoromethylmandelic acid and 25.4 g of HOBt into 300 mLDCM and stir to dissolve, pre-cool at 0°C for 10 min, add 15.2 g of DIC and stir for 10 min, then add resin, stir at room temperature for 2.5 h, filter , washed with DCM 6 times, that is.
[0079] The RinkAmide resin modified by RinkAmide resin and 4-trifluoromethylmandelic acid is carried out to infrared spectroscopic analysis, and the measured results are as follows: figure 1 shown.
[0080] Observed figure 1 , A represents the unmodified RinkAmide...
Embodiment 2
[0082] A preparation method of bremelanotide resin, said method comprising:
[0083] 1) Preparation of Fmoc-Lys(Boc)-resin:
[0084] Using the RinkAmide resin modified with 4-trifluoromethylmandelic acid obtained in Example 1 as a carrier, the resin was added to the piperidine / DCM mixed solution (the mass percentage of piperidine was 25%) and reacted for 20 min, rinsed with DCM 5 times; mix 70.2g Fmoc-Lys(Boc)-OH with 36g HOBt, 300 mLDCM, stir to dissolve, pre-cool at 0°C for 10 minutes, add 21.5g DIC and stir for 8 minutes, then add to the resin, stir and react for 10 minutes , add 2 gDMAP, stir and react at room temperature for 3.5h, add 46 g acetic anhydride, 35.5 g pyridine, react at room temperature for 40 min, wash 6 times with DCM, and obtain Fmoc-Lys(Boc)-resin shown in formula (1) ,
[0085] (1);
[0086] 2) Preparation of Fmoc-Trp(Boc)-Lys(Boc)-resin:
[0087] The Fmoc-Lys(Boc)-resin obtained in step 1) was added to the piperidine / DCM mixed solution (the mass ...
Embodiment 3
[0110] A kind of preparation method of bremelanotide crude product:
[0111] 1) Cracking: Mix TFA:TIS:deionized water according to the mass ratio of 93:3:4 to obtain a cracking agent, pre-cool at -20°C for 1 h, add the bremelanotide resin obtained in Example 2 to 1 L cracking agent solution, reacted at 0°C for 1 h, raised to room temperature and reacted for 3 h to remove the resin, filtered off the resin, and added the filtrate to 14 L pre-frozen methyl tert-butyl ether under stirring conditions, a large amount of white matter was precipitated, centrifuged, and collected The resulting solid was centrifuged, washed with methyl tert-butyl ether until neutral, and dried;
[0112] 2) Ester bond hydrolysis: Step 1) Dissolve the product in 20wt% acetonitrile aqueous solution, adjust pH=12 with 1 mol / L NaOH aqueous solution under ice-bath stirring condition, react at 0°C for 0.5 h, raise to 30°C to continue reaction 1 h, centrifuge to remove the precipitated white substance, adjust ...
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