Preparation method of abaloparatide

An abaparatide and synthesis method technology, which is applied in the field of polypeptide drug preparation, can solve the problems of increasing the difficulty of product separation and purification, low condensation efficiency, increased production cost and the like, and achieves improved synthesis efficiency, low consumption of raw materials and less impurities. Effect

Inactive Publication Date: 2018-05-18
润辉生物技术(威海)有限公司
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  • Summary
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AI Technical Summary

Problems solved by technology

However, this method requires a freeze-drying process before peptide fragments 2 and 3 are condensed, and the generation of peptide resin fragments 2 and 3 requires the use of a large amount of expensive 2-CTC resin; at the same time, the molar ratio of amino acid to resin dosage is 3 during amino acid condensation :1, when the peptide fragments are condensed, the molar ratio of peptide fragments 2 and 3 to the amount of peptide resin fragment 1 is 5:1, resulting in the consumption of a large amount of raw materials, resulting in increased production costs
[0010] Arg is a difficult amino acid in the solid-phase synthesis reaction, and the condensation efficiency is low during the condensation process, and it is very easy to produce [-Arg] missing peptides, while the abaloparatide sequence contains 3 consecutive Arg residues, which is very easy to produce Deletion of peptide impurities [-1Arg]-Abaloparatide, [-2Arg]-Abaloparatide and [-3Arg]-Abaloparatide, and even abaloparatide impurities with simultaneous deletion of multiple Args
At the same time, due to their own structural characteristics, Gly and Ala are easy to repeatedly condense during the condensation process, and it is easy to produce [+1Gly]-Abaloparatide and [+1Ala]-Abaloparatide impurities, these impurities and the target peptide The polarity is similar, it is not easy to separate, which increases the difficulty of product separation and purification

Method used

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  • Preparation method of abaloparatide
  • Preparation method of abaloparatide

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Preparation of dipeptide fragment Fmoc-Thr(OtBu)-Ala-OH:

[0046] 1) Synthesis of Fmoc-Thr(OtBu)-OSu:

[0047] Dissolve 0.1mol Fmoc-Thr(OtBu)-OH and 0.12mol HOSu in 0.2LTHF, and place in an ice-water bath to obtain mixed solution I; dissolve 0.12mol DCC in 0.1LTHF, and drop them into mixed solution I, and the addition is complete After continuing the reaction for 1 hour, the temperature was raised to 25°C. After 3 hours of heat-retaining reaction, the reaction solution was filtered and evaporated to dryness, then dissolved in DCM, filtered, and evaporated to dryness to obtain solid I; Crystallization afforded Fmoc-Thr(OtBu)-OSu.

[0048] 2) Synthesis of Fmoc-Thr(OtBu)-Ala-OH:

[0049] 0.15mol H-Ala-OH and 0.15mol Na 2 CO 3 Dissolve in 0.2L volume fraction 50%THF-H 2 O solution to obtain mixed solution II; Fmoc-Thr(OtBu)-OSu was dissolved in THF and added dropwise to mixed solution II. After overnight reaction at 25°C, the reaction solution was rotary evaporated and...

Embodiment 2

[0051] Preparation of dipeptide fragment Fmoc-Lys(Boc)-Gly-OH:

[0052] 1) Synthesis of Fmoc-Lys(Boc)-OSu:

[0053] Dissolve 0.1mol Fmoc-Lys(Boc)-OH and 0.12mol HOSu in 0.2L THF, and place in an ice-water bath to obtain mixed solution I; dissolve 0.12mol DCC in 0.1LTHF, and drop them into mixed solution I, dropwise After completing the reaction for 1 hour, raise the temperature to 25°C, keep the temperature for 3 hours, filter the reaction solution, evaporate to dryness, add DCM to dissolve, filter, and evaporate to dryness to obtain solid I; add ethyl acetate to dissolve solid I, Recrystallization was performed to obtain Fmoc-Lys(Boc)-OSu.

[0054] 2) Synthesis of Fmoc-Lys(Boc)-Gly-OH:

[0055] 0.15mol H-Gly-OH and 0.15mol Na 2 CO 3 Dissolve in 0.2L volume fraction 50%THF-H 2 O solution to obtain mixed solution II; Fmoc-Lys(Boc)-OSu was dissolved in THF and added dropwise to mixed solution II, after overnight reaction at 25°C, the reaction solution was rotary evaporated,...

Embodiment 3

[0057] Preparation of Fmoc-Arg(Pbf)-Arg(Pbf)-Arg(Pbf)-Glu(OtBu)-OH:

[0058] 1) Synthesis of Fmoc-Arg(Pbf)-OSu:

[0059] Dissolve 0.1 mol Fmoc-Arg(Pbf)-OH and 0.12 mol HOSu in 0.2 L THF, and place in an ice-water bath to obtain mixed solution I; dissolve 0.12 mol DCC in 0.1 L THF, and add dropwise to mixed solution I, After the dropwise addition was completed and the reaction was continued for 1 hour, the temperature was raised to 25°C, and after 3 hours of heat preservation reaction, the reaction solution was filtered and evaporated to dryness, then dissolved in DCM, filtered, and evaporated to dryness to obtain solid I; add ethyl acetate to dissolve solid I , and recrystallized to obtain Fmoc-Arg(Pbf)-OSu.

[0060] 2) Synthesis of Fmoc-Arg(Pbf)-Arg(Pbf)-OH:

[0061] 0.15mol H-Arg(Pbf)-OH and 0.15molNa 2 CO 3 Dissolve in 0.2L volume fraction 50%THF-H 2 O solution to obtain mixed solution II; Fmoc-Arg(Pbf)-OSu was dissolved in THF and added dropwise to mixed solution II. ...

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Abstract

The invention relates to a preparation method of abaloparatide, aiming at solving the technical problems of a method in the prior art for synthesizing the abaloparatide that steps are complicated, theconsumption of raw materials is great and a synthesized product has low yield, low purity and more impurities and is difficult to purify. The preparation method of the abaloparatide, provided by theinvention, comprises the following steps: firstly, simultaneously synthesizing dipeptide segments Fmoc-Thr(OtBu)-Ala-OH, Fmoc-Lys(Boc)-Gly-OH and Fmoc-Ala-Val-OH and a tetrapeptide segment Fmoc-Arg-(Pbf)-Arg(Pbf)-Arg(Pbf)-Glu(OtBu)-OH; taking amino resin as a carrier of solid-phase synthesis; condensing completely protected amino acids containing alpha-NH2 with an Fmoc protection group and the synthesized dipeptide segments and tetrapeptide segment in sequence according to an amino acid sequence of the abaloparatide from a C end to an N end, so as to obtain completely protected abaloparatide resin, wherein 6 steps of solid-phase condensation reaction are reduced; cracking and purifying to obtain an abaloparatide pure product. The preparation method provided by the invention is widely applied to the technical field of preparation and synthesis of polypeptide drugs.

Description

technical field [0001] The invention belongs to the technical field of polypeptide drug preparation methods, and in particular relates to a preparation method of abaloparatide. Background technique [0002] Parathyroid hormone (PTH) is a polypeptide hormone secreted by parathyroid chief cells that regulates calcium and phosphorus metabolism in the body, and its C-terminal peptide chain binds to the PTH-II receptor to promote bone cell apoptosis, N Binding of telopeptide chains to PTH-I receptors promotes bone remodeling. Teriparatide, a drug for the treatment of osteoporosis developed by Lilly Company based on the N-terminal 34 amino acid residues of human parathyroid hormone (hPTH), was approved for marketing by the FDA in December 2002. [0003] Abaloparatide, the English name is Abaloparatide, is a new type of parathyroid hormone-related peptide (PTHrP) developed by Radius Health, which is a powerful selective activator of PTH-I receptor, which can increase bone mineral ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/635C07K1/20C07K1/06C07K1/02
CPCY02P20/55C07K14/635
Inventor 姬胜利殷金岗陈明鲁王广胜
Owner 润辉生物技术(威海)有限公司
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