Preparation method of liraglutide

A technology of liraglutide and synthetic method, which is applied in the field of preparation of polypeptide drugs, can solve the problems of many impurities, low yield, and unsuitability for industrial production

CN103980358AActive Publication Date: 2014-08-13ADLAI NORTYE BIOPHARMA CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2014-08-13

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Abstract

The invention relates to a synthesis technology of liraglutide, and solves the problems of long synthesis period, high cost, low yield and a large amount of impurity in the prior art. The method provided by the invention comprises the specific steps of: A) synthesizing a Fmoc-Lys-(Glu(N alpha-Palmitoyl)-OtBu)-OH fragment by a liquid phase; B) in the presence of an activating agent system, coupling a resin solid phase carrier and Fmoc-Gly-OH to obtain Fmoc-Gly-resin; C) successively coupling amino acids with N-terminal Fmoc protection and side chain protection according to peptide sequence of liraglutide backbone by a solid phase synthesis method, wherein the lysine tripeptide fragment employs Fmoc-Lys-(Glu(N alpha-Palmitoyl)-OtBu)-OH; and D) cracking, purifying and freeze-drying to obtain liraglutide. The invention provides a liraglutide synthesis technology with short synthesis cycle, low cost, high yield and suitability for scale production.
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Description

Technical field

[0001] The present invention involves a preparation method of a polypeptide drug. It is a synthetic long-acting II diabetes with a synthesis of cricket-1 (GLP-1) receptor agonist-Lillawhan peptidePreparation method. Background technique

[0002] Leralugin, the English name: liraglutide, the structure is as follows:

[0003]

[0004] The peptide sequence is:

[0005] H-His-Ala-GLU-GLY-ThR-PHR-Ser-Val-Ser-Ser-Leu-GLN-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA-ALA (N N-N-N-N-N-N-(n N-n-n-(n N-n-N-N-N-N-Palmitoyl-l-γ-glutamyl))-Glu-PHE-DRA-ALA-Leu-Val-Gly-Gly-Gly-OH

[0006] The first and only long-acting GLP-1 analogs developed by Dannino and Nond Corporation are currently the only long-acting GLP-1 analog.Immunism is similar to GLP-1.The molecular structure and GLP-1 (7-37) of the Liculugin peptide reached 97 %. 34 Replaced by ARG, LYS 26 Palmlycinomacinomacinomacinomacinomaci...

Examples

Embodiment 1

[0091] Example 1: Palmitoyl-OSU activation fat synthesis

[0092] Called 256.42g positive hexisc acid (1.0 mol), 138.10g HOSU (1.2mol) Add 2000ml THF, add 247.56g DCC (1.2mol) under the ice water bath, react for 1 hour, react to room temperature for 3 hours, and response for 3 hours.Filter, dry the mother liquid, dissolve with DCM, filtrate, washed sodium saturated sodium bicarbonate 3 times, 2 times of pure water, 2 times of anti-extract, combined with organic phase, dry sodium carbonate, dried, 0-5 ° C ice ethanol, 0-5 ° C ice ethanol3 times heavy crystal, filtering, solid oil pump to get 314.62G Palmitoyl-Osu activated lipids, with 89%income.

Embodiment 2

[0093] Example 2: Palmitoyl-Glu-OTBU synthesis

[0094] It is said that 101.62g H-Glu-OTBU (0.5mol) and 79.50G NA 2 CO 3 (0.75mol) Add to a mixed solution of 500ml of water and 500ml THF to dissolve, which is called 176.75G Palmitoyl-OSU (0.5mol) and adds to 500ml THF.Dilute hydrochloric acid regulate pH to 7, rotate the THF, and then adjust the pH to 3.Get a lot of white precipitation and filter.The obtained white precipitation is required with 0-5 ℃ ice ethanol.The solid oil pump was pulled by 192.11g Palmitoy-Glu-OTBU, with a revenue of 87%.

Embodiment 3

[0095] Example 3: Palmitoyl-Glu (OSU) -s' synthesis of OTBU

[0096] Called 88.33G Palmitoyl-Glu-OTBU (0.2mol), 27.62g HOSU (0.24mol) Add 1000ml THF, add 49.51g DCC (0.24mol) under the ice water bath, respond to 1 hour, heating to room temperature for 3 hours, reaction to the reaction.Filter, dry the mother liquid, dissolve with DCM, filtrate, washed sodium saturated sodium bicarbonate 3 times, 2 times of pure water, 2 times of anti-extract, combined with organic phase, dry sodium carbonate, dried, 0-5 ° C ice ethanol, 0-5 ° C ice ethanol3 times heavy crystal, filtering, solid oil pump to get 94.81g Palmitoyl-Glu (OSU) -ebu activated lipid with 88%income.