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

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 ...

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 require very harsh storage conditions. Once stored improperly, not only seriously affect The modification efficiency of protein and polypeptide drugs, and affects the entire production process, brings many uncertain factors to the industrialization of this technology

For example, polyethylene glycol acetaldehyde has high reactivity, and it is very easy to undergo aldol condensation reaction in preparation and storage to cause by-products. Product quality and storage and use are difficult to be effectively controlled. Therefore, it has not been well obtained. development and use of

Chinese patent CN200610099081.0 discloses that amide bonds are used to replace alkoxy groups, so that polyethylene glycol acetaldehyde derivatives have higher stability, but the derivatives still have higher activity of aldehyde groups and There are unstable factors

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