Preparation method for synthesizing nesiritide through solid-liquid combination

A technology of nesiritide and solid-phase synthesis, which is applied in the field of preparation of solid-liquid combination synthesis of nesiritide, can solve the problem of affecting product yield, difficult to avoid the formation of defective Ser impurities or racemic impurities, and high material cost question

Pending Publication Date: 2021-03-19
HYBIO PHARMA
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] 1. There is Ser in the peptide sequence which is difficult to couple 19 -Ser-Ser-Ser 22 Fragments, it is difficult to avoid the formation of defective Ser impurities or racemic impurities by using conventional solid-phase step-by-step coupling synthesis methods;
[0011] 2. The solid-phase fragment synthesis method can reduce the formation of defective impurities or racemized impurities, but peptide fragments require multiple feeds, and the material cost is very high;
[0012] 3. Using solid phase I 2 The method of oxidation will lead to the shedding of the fully protected peptide and the formation of other impurities, which will affect the yield of the final product

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation method for synthesizing nesiritide through solid-liquid combination
  • Preparation method for synthesizing nesiritide through solid-liquid combination
  • Preparation method for synthesizing nesiritide through solid-liquid combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] The synthesis of embodiment 1, Fmoc-His (Trt)-2CTC resin

[0066] Weigh 100 g of 2-CTC resin (purchased from Tianjin Nankai Synthetic Technology Co., Ltd.) with a degree of substitution of 0.8 mmol / g, add it to a solid-phase reaction column, wash it twice with DMF, and swell the resin with DMF for 30 minutes, then take 49.2 g Fmoc-His(Trt)-OH (80mmol) was dissolved in 320ml of DMF, activated by adding 27.2ml (80mmol) of DIPEA at -10°C to 0°C, and then added to the above-mentioned reaction column equipped with resin. After reacting for 2 hours, add Block with 40 mL of anhydrous methanol for 1 hour. Wash 3 times with DMF and 3 times with DCM, then block with anhydrous methanol for 30 minutes, shrink and dry with methanol to obtain 140 g of Fmoc-His(Trt)-2CTC resin, and the detection degree of substitution is 0.452 mmol / g.

Embodiment 2

[0067] Embodiment 2, the synthesis of intermediate 1

[0068] Weigh the Fmoc-His(Trt)-2CTC 44g (20mmol) with a degree of substitution of 0.452mmol / g prepared in Example 1, add it to the solid-phase reaction column, wash it twice with an appropriate amount of DMF, swell with DMF for 30 minutes, and wash it with DBLK Remove Fmoc protection, then wash with DMF for 4 times and DCM for 2 times, and detect the color of the resin with the ninhydrin method. If the resin has color, it means that Fmoc has been removed. Take 39.0g Fmoc-Arg(Pbf)-OH (60mmol), 9.8g HOBt (72mmol), dissolve in 120ml of DCM and DMF mixed solution with a volume ratio of 1:1, add 12.1mL DIC ( 72mmol) was activated for 3 minutes and added to the solid-phase reaction column, and reacted at room temperature for 2 hours. Use the ninhydrin method to detect and judge the reaction end point. If the resin is colorless and transparent, it means that the reaction is complete.

[0069] Repeat the above steps of removing ...

Embodiment 3

[0071] The synthesis of embodiment 3, Fmoc-Ser (tBu)-2CTC resin

[0072] Weigh 100 g of 2-CTC resin (purchased from Tianjin Nankai Synthetic Technology Co., Ltd.) with a degree of substitution of 0.8 mmol / g, add it to a solid-phase reaction column, wash it twice with DMF, and swell the resin with DMF for 30 minutes, then take 46.0 g Fmoc-Ser(tBu)-OH (120mmol) was dissolved in 320ml of DMF, activated by adding 10.8ml (120mmol) of DIPEA at -10°C to 0°C, and then added to the above-mentioned reaction column filled with resin. After reacting for 2 hours, add Block with 40 mL of anhydrous methanol for 1 hour. Wash 3 times with DMF and 3 times with DCM, then block with anhydrous methanol for 30 minutes, shrink and dry with methanol to obtain 126 g of Fmoc-Ser(tBu)-2CTC resin, and the detection degree of substitution is 0.515 mmol / g.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a preparation method of nesiritide. The preparation method comprises the following steps of 1) synthesizing an intermediate 1 by a solid-phase synthesis method; 2) synthesizing an intermediate 2 by a solid-phase synthesis method, activating the C-terminal carboxyl of the intermediate 2 to obtain an intermediate 3, and removing a side chain protecting group from the intermediate 3 to obtain an intermediate 4; 3) performing disulfide bond exchange reaction on the intermediate 1 and the intermediate 4 to form an intermolecular disulfide bond intermediate 5; 4) enabling the intermediate 5 to form an amido bond under the action of a cyclization promoting reagent to obtain an intermediate 6, and then removing an Fmoc protecting group to obtain a nesiritide crude product;and 5) purifying the nesiritide crude product to obtain the nesiritide. A liquid phase and solid phase combined method is adopted, so that the preparation efficiency is high, and the purity is high.

Description

technical field [0001] The invention relates to the field of drug synthesis, in particular to a preparation method for synthesizing nesiritide through solid-liquid combination. Background technique [0002] Nesiritide is a class of anti-cardiac insufficiency drugs. It is a B-type natriuretic peptide synthesized by recombinant DNA technology. By binding to natriuretic titanium receptors. Nesiritide has clear vasodilator and natriuretic effects. Clinical studies have shown that this is a new drug for the treatment of acute heart failure with significant efficacy, quick onset and few adverse reactions, and it can improve the hemodynamics and clinical symptoms of heart failure patients. [0003] Nesiritide, the English name is nesiritide, the molecular weight is 3464.04, and the molecular formula is: Ser 1 -Pro-Lys-Met-Val-Gln-Gly-Ser-Gly-Cys 10 -Phe-Gly-Arg-Lys-Met-Asp-Arg-Ile-Ser-Ser 20 -Ser-Ser-Gly-Leu-Gly-Cys-Lys-Val-Leu-Arg 30 -Arg-His (Disulfide Bridge: Cys10-Cys26)...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K14/58C07K1/16C07K1/08C07K1/06C07K1/04C07K1/02
CPCC07K14/58C07K7/06C07K1/04C07K1/06Y02P20/55
Inventor陈永汉尹传龙陶安进袁建成
OwnerHYBIO PHARMA