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Preparation method of polyethylene glycol loxenatide based on reaction kinetics

A reaction kinetics, polyethylene glycol technology, applied in the fields of bioengineering and pharmaceutical engineering, can solve the problems of difficult optimization and control, many influencing factors, etc., to reduce the probability of modifying amino groups, reduce degradation or polymerization, and increase The effect of recovery

Inactive Publication Date: 2019-06-25
JIANGSU HANSOH PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to many influencing factors, conventional experimental methods are used for mPEG 2 -MAL (40kD) modified loxenatide reaction process is still very difficult to optimize and control

Method used

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  • Preparation method of polyethylene glycol loxenatide based on reaction kinetics
  • Preparation method of polyethylene glycol loxenatide based on reaction kinetics
  • Preparation method of polyethylene glycol loxenatide based on reaction kinetics

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0084] 3. Preparation of PEG Loxenatide

[0085] (1) Synthesis of polyethylene glycol loxenatide

[0086] Weigh 3g loxenatide and a certain amount of mPEG respectively 2 -MAL is dissolved in 1L 0.1mol / L sodium phosphate buffer for reaction. The reaction conditions are as follows: the pH of the sodium phosphate buffer solution is 6.3-7.0, the temperature is 15-30℃, and mPEG 2 -The molar ratio of MAL / Losenatide is 1.10:1-1.15:1, and the reaction time is 15-90min; after the reaction, a crude polyethylene glycol losenatide solution is obtained;

[0087] (2) Purification of polyethylene glycol loxenatide

[0088] The crude polyethylene glycol loxenatide solution obtained in step (1) was purified by cation exchange chromatography to obtain the polyethylene glycol loxenatide salt solution; the polyethylene glycol was desalted by RP-HPLC, concentrated under reduced pressure, and lyophilized to obtain polyethylene glycol. Loxenatide glycol pure product.

[0089] Wherein, the cation exchange ch...

Embodiment 1

[0110] Example 1 mPEG based on reaction kinetics 2 -MAL (40kD) modified loxenatide process optimization control technology (small scale)

[0111] Follow the method in 2 above to establish mPEG based on reaction kinetics 2 -MAL (40kD) modified losenatide process optimization control technology. Among them, the pH of the sodium phosphate buffer is 5.5, 6.3, 7.0, and 7.7, and the temperature is 15, 20, 25, and 30°C. mPEG 2 -The molar ratio of MAL / Losenatide is 0.92:1, 1.15:1, 1.27:1 and 1.38:1, and the reaction time is 0-120min. Take sodium phosphate buffer pH 6.3, temperature 20℃, mPEG 2 -The reaction conditions with the molar ratio of MAL / Losenatide of 1.15:1 were used as a reference standard to analyze the influence of individual factors.

[0112] MPEG under different conditions 2 -MAL (40kD) modified losenatide reaction kinetics such as Figure 1-Figure 12 As shown, the kinetic parameters obtained by fitting the experimental data to the kinetic model are shown in Table 2.

[0113] ...

Embodiment 2 and comparative example 1

[0119] Example 2 and Comparative Example 1 Synthesis of polyethylene glycol losenatide (pilot scale)

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Abstract

The invention relates to a preparation method of polyethylene glycol loxenatide based on reaction kinetics. Especially, the invention relates to the preparation method of the polyethylene glycol loxenatide. The synthesis method is based on a process optimization control technology of reaction kinetics, can maximize the loxenatide and mPEG2-MAL (40 kD) material utilization rate, and reduces the occurrence of side reactions and the formation of impurities. The purification method uses cation exchange chromatography to obtain high-purity and high-recovery polyethylene glycol loxenatide. The process is simple and efficient, and is suitable for industrial production.

Description

Technical field [0001] The invention belongs to the fields of bioengineering and pharmaceutical engineering, and relates to polyethylene glycol modification of protein and polypeptide drugs, and specifically relates to a preparation method of polyethylene glycol losenatide, especially a synthesis and purification method. Background technique [0002] Diabetes is a global epidemic, and its annual death rate is second only to tumors and cardiovascular and cerebrovascular diseases. The main feature of diabetes is that the body is in a state of high blood sugar. According to the time and cause of diabetes, the International Diabetes Federation (IDF) divides diabetes into type 1 diabetes, type 2 diabetes and gestational diabetes. Among all diagnosed diabetic patients, more than 90% of diabetic patients are type 2 diabetic patients. Type 2 diabetes is characterized by insulin suppression and damage to pancreatic β-cell function, leading to insulin deficiency and hyperglycemia. Type ...

Claims

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Application Information

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IPC IPC(8): C07K14/575C07K1/107C07K1/18
Inventor 王旭东修志龙魏宁宁朱后田张丰盈袁恒立
Owner JIANGSU HANSOH PHARMA CO LTD
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