Preparation method for liraglutide

A technology of liraglutide and liraglutide, which is applied in the field of polypeptide drug preparation, can solve the problems of low total yield and purity of the product, difficulty in separation and purification of the preparation method, etc., and achieve low purification difficulty, wide practical value and application Prospects, the effect of improving product yield

Active Publication Date: 2013-09-04
CHENGDU SHENGNUO BIOPHARM
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Problems solved by technology

[0013] The technical problem to be solved by the present invention is that the existing preparatio

Method used

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  • Preparation method for liraglutide

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preparation example Construction

[0045] The preparation method of liraglutide comprises: adopting solid-phase polypeptide synthesis method to prepare liraglutide peptide resin, liraglutide peptide resin is subjected to acid hydrolysis to obtain liraglutide crude product, and finally liraglutide crude product is purified to obtain liraglutide peptide resin Laglutide pure product; wherein the steps of preparing liraglutide peptide resin by solid-phase peptide synthesis method are: on Fmoc-Gly-carrier resin, sequentially insert the corresponding protected amino acids in the following sequences or Fragment, preparation of liraglutide peptide resin:

[0046] Boc-W(Trt)-X(OtBu)-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-

[0047] Asp(OtBu)-Val-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-X(OtBu)-

[0048] Gln(Trt)-Ala-Ala-Lys[Y(α-OtBu)]-Glu(OtBu)-Phe-Ile-Ala-

[0049] Trp(Boc)-Leu-Val-Z(Pbf)-Arg(Pbf)-Gly-resin;

[0050] Wherein, W is His-Ala, X is Glu-Gly, Y is Nα-PAL-Glu, and Z is Arg-Gly.

[0051] When accessing W, one-step access met...

Embodiment 1

[0078] The synthesis of embodiment 1 liraglutide peptide resin

[0079] Liraglutide peptide resin is:

[0080] Boc-W(Trt)-X(OtBu)-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-

[0081] Asp(OtBu)-Val-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-X(OtBu)-

[0082] Gln(Trt)-Ala-Ala-Lys[Y(α-OtBu)]-Glu(OtBu)-Phe-Ile-Ala-

[0083]Trp(Boc)-Leu-Val-Z(Pbf)-Arg(Pbf)-Gly-resin;

[0084] Wherein, W is His-Ala, X is Glu-Gly, Y is Nα-PAL-Glu, and Z is Arg-Gly.

[0085] When inserting W, the corresponding protected amino acid is Boc-His(Trt)-Ala-OH; when inserting X, the corresponding protected amino acid is Fmoc-Glu(OtBu)-Ala-OH; when inserting Y, adopt two-step access method, the corresponding protected amino acids when Y is inserted in two steps are Fmoc-Glu(α-OtBu)-OH and PAL (palmitic acid); when Z is inserted, the corresponding protected amino acids are Fmoc-Arg(Pbf)-Gly-OH; The protected amino acid used when inserting Lys is Fmoc-Lys (Alloc).

[0086] Using Fmoc-Gly-resin as the initial carrier, the liraglu...

Embodiment 2

[0102] Example 2 Preparation of Liraglutide Crude Product

[0103] Take the liraglutide peptide resin prepared in Example 1, add a lysis reagent with a volume ratio of TFA:water:EDT=95:5:5 (lysis reagent 10mL / g resin), stir evenly, and stir at room temperature for 3 hours, The reaction mixture was filtered with a sand core funnel, the filtrate was collected, the resin was washed 3 times with a small amount of TFA, the combined filtrates were concentrated under reduced pressure, anhydrous ether was added to precipitate, and anhydrous ether was used to wash the precipitate 3 times, and the off-white powder was obtained by draining. It is the crude product of liraglutide, and the purity of the crude product is 45.3%.

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Abstract

The invention belongs to the technical field of preparation methods for polypeptide medicines, and particularly relates to a preparation method for liraglutide, aiming at solving the technical problems of difficult separation and purification, and low total yield and purity of products, of the existing preparation method. The technical scheme for solving the technical problems, disclosed by the invention is providing a preparation method for liraglutide. The method comprises the following steps of: preparing liraglutide resin via a solid-phase polypeptide method; performing acidolysis to obtain a liraglutide crude product; and finally purifying to obtain a liraglutide pure product, wherein the solid-phase polypeptide method comprises the step of sequentially connecting amino resin to corresponding protected amino acids or protected amino acid segments in the following sequences via a solid-phase coupling synthesis method, so as to prepare the liraglutide resin: Boc-W(Trt)-X(OtBu)-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Val-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-X(OtBu)-Gln(Trt) -Ala-Ala-Lys[Y(Alpha-OtBu)]-Glu(OtBu) -Phe-Ile-Ala-Trp(Boc)-Leu-Val-Z(Pbf)-Arg(Pbf)-Gly-resin, wherein W is His-Ala, X is Glu-Gly, Y is N alpha-PAL-Glu, and Z is Arg-Gly. The invention provides a novel method for shortening the production cycle, and improving the purity and the yield of the products.

Description

technical field [0001] The invention belongs to the technical field of polypeptide drug preparation methods, in particular to a preparation method of liraglutide. Background technique [0002] Liraglutide has the following structure: [0003] His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser- [0004] Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Nα-PAL-γ-Glu)- [0005] Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-OH. [0006] Liraglutide is a GLP-1 (glucagon-like peptide) analog with 97% sequence homology to human GLP-1, which can bind and activate the GLP-1 receptor. The GLP-1 receptor is the target of natural GLP-1, an endogenous incretin hormone that promotes glucose-dependent insulin secretion from pancreatic β-cells. Different from Natural GLP-1, the pharmacokinetic and pharmacodynamic characteristics of liraglutide in humans are suitable for once-daily dosing regimen. After subcutaneous injection, the mechanism of prolonged action includes: self-association to slow absorption; bi...

Claims

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Application Information

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IPC IPC(8): C07K14/605C07K1/16C07K1/06C07K1/04
Inventor 郭德文曾德志卢昌亮文永均
Owner CHENGDU SHENGNUO BIOPHARM
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