Protein polypeptide drug N-terminal fixed-point polyethylene glycol modification method

A polyethylene glycol, protein polypeptide technology, applied in the preparation methods of peptides, chemical instruments and methods, peptides, etc., can solve the problems of difficult to effectively control product quality, storage and use, affecting the production process, and expensive raw materials. Conducive to storage for backup, avoids easy failure, and has the effect of convenient source

Active Publication Date: 2015-09-23
ZHEJIANG PHARMA COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The oxidation method is obtained by directly oxidizing the terminal hydroxyl group of the PEG chain, for example, adding oxygen to the mixture of PEG and catalyst to oxidize the terminal hydroxyl group of the PEG chain into an aldehyde group, or using MnO 2 etc. as oxidants directly oxidize the terminal hydroxyl groups, but in most cases, these oxidation conditions easily lead to the decomposition of PEG chains
The acetal method is obtained by introducing acetal groups at the ends of PEG chains followed by hydrolysis, but the reactant raw materials used in this method are relatively expensive and not suitable for commercialization
[0005] In addition, many existing PEG modifiers, especially PEG aldehyde derivatives, have poor stability due to their active groups. In order to ensure the modification efficiency during use, they often req...

Method used

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  • Protein polypeptide drug N-terminal fixed-point polyethylene glycol modification method
  • Protein polypeptide drug N-terminal fixed-point polyethylene glycol modification method
  • Protein polypeptide drug N-terminal fixed-point polyethylene glycol modification method

Examples

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Embodiment 1

[0031] The method for N-terminal site-directed polyethylene glycol modification of protein polypeptide drugs in this embodiment includes the following steps:

[0032] (1) Change the terminal hydroxyl group of PEG into a group with higher activity to obtain a PEG active intermediate, and then use the PEG active intermediate to combine with a small group of o-hydroxyl, α-hydroxy acid, β-amino alcohol or α-amino ketone structure Molecular compounds are reacted to obtain polyethylene glycol intermediates; specifically, the PEG active intermediates used in this example are prepared by converting the terminal hydroxyl groups of PEG with single-end blocking into groups with higher activity. In this embodiment, monomethoxypolyethylene glycol (mPEG5000, with an average relative molecular weight of 5000) was chosen. The terminal hydroxyl group of mPEG5000 is converted into p-nitrophenyl carbonate to obtain PEG active intermediate, and then reacted with small molecular compound 4-amino-2...

Embodiment 2

[0044] The method for N-terminal site-directed polyethylene glycol modification of protein polypeptide drugs in this embodiment includes the following steps:

[0045] (1) The PEG active intermediate selected in the present embodiment is double-chain polyethylene glycol lysine succinimide ester (2mPEG5000-Lys-NHS, the average relative molecular weight of each polyethylene glycol chain is 5000), and It reacts with the small molecular compound 3-amino-1,2-propanediol to obtain the polyethylene glycol intermediate 2mPEG5000-Lys-o-diol, whose structural formula is Wherein n is 113 on average, m is 1, k is 2, R is a methoxy group, X is an amide bond, Y is an adjacent hydroxyl structure, and Z is lysine (Lys).

[0046] (2) The polyethylene glycol intermediate 2mPEG5000-Lys-o-diol is oxidized with sodium periodate into polyethylene glycol aldehyde derivatives with active aldehyde groups at the end, which can be used for protein and polypeptide drugs after dialysis The fixed-point po...

Embodiment 3

[0055] The method for N-terminal site-directed polyethylene glycol modification of protein polypeptide drugs in this embodiment includes the following steps:

[0056] (1) The PEG active intermediate selected in this embodiment is monomethoxypolyethylene glycol amino hydrochloride (mPEG20000-NH 2 HCl, the average relative molecular weight is 20000), react it with the small molecule compound N-tert-butoxycarbonyl-L-serine, and then remove the Boc group to obtain the polyethylene glycol intermediate monomethoxypolyethylene glycol Serine (mPEG20000-Ser), its structural formula is Wherein the average n is 454, m is 0, R is a methoxyl group, X is an amide bond, and Y is a β-aminoalcohol structure.

[0057] (2) The polyethylene glycol intermediate mPEG20000-Ser is oxidized with sodium periodate into polyethylene glycol aldehyde derivatives with active aldehyde groups at the end, which can be used for the fixed-point polymerization of the N-terminus of protein and polypeptide drugs ...

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Abstract

The invention discloses a protein polypeptide drug N-terminal fixed-point polyethylene glycol modification method. The method comprises the steps that 1, PEG terminal hydroxyl groups are converted into groups with higher activity to obtain a PEG active intermediate, and the PEG active intermediate and a small molecule compound of a hydroxyphenylacetic acid or alpha-hydroxy acid or beta-amino alcohol or alpha-aminoketones structure react to obtain a polyethylene glycol intermediate; 2, the polyethylene glycol intermediate is oxidized by an oxidizing agent to be a polyethylene glycol aldehyde derivative with active aldehyde groups at the terminal, then the polyethylene glycol aldehyde derivative is used for protein polypeptide drug N-terminal fixed-point polyethylene glycol modification, pegylation protein polypeptide drugs of a stable structure can be obtained through the reductive amination effect of a reducing agent, and the polyethylene glycol activity modifying agent preparation and decoration of the polyethylene glycol activity modifying agent for the protein polypeptide drugs need to be tightly connected. According to the protein polypeptide drug N-terminal fixed-point polyethylene glycol modification method, the problem that many polyethylene glycol aldehyde derivatives are likely to lose efficacy due to improper storage can be avoided, the period is short, and the cost is low.

Description

technical field [0001] The invention relates to a method for modifying the N-terminus of protein and polypeptide drugs with fixed-point polyethylene glycol. Background technique [0002] Polyethylene glycol (PEG) is an extremely versatile synthetic polymer compound, which is mainly used in many fields such as biomedicine, chemical industry, food, and material science. PEG is easily soluble in water and organic solvents, and has good physical and chemical properties and biocompatibility. In biomedicine, PEG is widely used as an excipient in pharmaceutical preparations, sustained-release and targeting studies because of its amphiphilicity, good safety, biocompatibility, inactivity, and no toxic or side effects. It can also be covalently linked to proteins or polypeptides to improve the biochemical and pharmacokinetic properties of proteins and polypeptide drugs, and even for the modification of small molecule drugs. A large number of studies have shown that the antigenicity ...

Claims

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Application Information

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IPC IPC(8): C07K1/107
Inventor 陈阳建
Owner ZHEJIANG PHARMA COLLEGE
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