阅读说明：本技术 一种二氢噻唑杂环化合物的合成方法及在生物分子修饰中的应用 (Synthesis method of dihydrothiazole heterocyclic compound and application of dihydrothiazole heterocyclic compound in modification of biological molecules ) 是由 吴川六 李卓儒 高巍 郑晓莉 于 2019-10-25 设计创作，主要内容包括：一种二氢噻唑杂环化合物的合成方法及在生物分子修饰中的应用,涉及有机合成技术领域。将硫烯醚化合物和含1,2-巯基乙胺骨架结构化合物在溶剂中混合,加入弱碱进行反应,反应温度为20～60℃,反应时间为60～240min,即得到二氢噻唑杂环化合物。所述二氢噻唑杂环化合物的合成方法,温和、高效,可以在室温及水中快速反应,克服传统合成方法中反应条件剧烈、时间长等缺点；利用该合成方法,可以高效、精准地对含有氮端半胱氨酸残基的生物分子如多肽及蛋白进行修饰,并进一步开发检测和治疗方法等应用。(A synthetic method of a dihydrothiazole heterocyclic compound and application thereof in biomolecule modification, relating to the technical field of organic synthesis. Mixing a thioether compound and a compound containing a 1, 2-mercaptoethylamine skeleton structure in a solvent, adding a weak base to react at the temperature of 20-60 ℃ for 60-240 min, and obtaining the dihydrothiazole heterocyclic compound. The synthesis method of the dihydrothiazole heterocyclic compound is mild and efficient, can quickly react at room temperature and in water, and overcomes the defects of violent reaction conditions, long time and the like in the traditional synthesis method; by utilizing the synthesis method, biomolecules containing the nitrogen-terminal cysteine residue, such as polypeptide and protein, can be modified efficiently and accurately, and further, the detection and treatment methods and other applications can be developed.)

1. A method for synthesizing a dihydrothiazole heterocyclic compound is characterized by comprising the following steps: mixing a thioether compound and a compound containing a 1, 2-mercaptoethylamine skeleton structure in a solvent, and adding weak base to react to obtain the dihydrothiazole heterocyclic compound.

2. A process for the synthesis of a dihydrothiazole heterocyclic compound according to claim 1, characterized in that: the reaction temperature is 20-60 ℃, and the reaction time is 60-240 min.

3. A process for the synthesis of a dihydrothiazole heterocyclic compound according to claim 1, characterized in that: the thiolene ether compound has the following structural general formula:

wherein X is sulfur or oxygen, R₁Is an alkyl chain, alkoxy chain, cycloalkyl or aryl, optionally substituted or unsubstituted, except for a mercapto group, amino substituent; r₂Is phenyl optionally substituted or unsubstituted except for mercapto, amino substituents;

the compound containing the 1, 2-mercaptoethylamine framework structure has the following structural general formula:

wherein R is₃Is hydrogen or an optionally substituted group.

4. A process for the synthesis of a dihydrothiazole heterocyclic compound according to claim 1, characterized in that: the molar ratio of the thiolene ether compound to the compound containing the 1, 2-mercaptoethylamine skeleton structure is 1 (1-2).

5. A process for the synthesis of a dihydrothiazole heterocyclic compound according to claim 4, characterized in that: the mol ratio of the thiolene ether compound to the compound containing the 1, 2-mercaptoethylamine skeleton structure is 1: 1.

6. A process for the synthesis of a dihydrothiazole heterocyclic compound according to claim 1, characterized in that: the solvent comprises at least one of water, methanol, ethanol and acetonitrile; the weak base comprises at least one of sodium carbonate, sodium bicarbonate, potassium carbonate, phosphate buffer and triethylamine.

7. A process for the synthesis of a dihydrothiazole heterocyclic compound according to claim 6, characterized in that: the solvent is a mixed solvent of water and acetonitrile, and the volume ratio is 1: 1; the weak base is sodium bicarbonate.

8. The application of the synthetic method of the dihydrothiazole heterocyclic compound in the modification of biological molecules is characterized in that: mixing and reacting a thioether compound and a biomolecule containing cysteine residue at the nitrogen end in a weak base environment to obtain the modified biomolecule connected by a dihydrothiazole heterocyclic skeleton.

9. The use of a method of claim 8 for the synthesis of a dihydrothiazole heterocyclic compound for the modification of biomolecules, wherein: the structural general formula of the thioether compound is as follows:

wherein X is sulfur or oxygen, R₁Is an alkyl chain, alkoxy chain, cycloalkyl or aryl, optionally substituted or unsubstituted, except for a mercapto group, amino substituent; r₂Is phenyl optionally substituted or unsubstituted except for mercapto, amino substituents; y is covalently bonded to R₂Connected byDyes, drugs, functional biomolecules; the molar ratio of the thioether compound to the biomolecule to be modified is (1-100): 1.

10. The use of a method of claim 8 for the synthesis of a dihydrothiazole heterocyclic compound for the modification of biomolecules, wherein: the weak base is phosphoric acid buffer solution, the pH value is 7.4, the reaction temperature is 25-37 ℃, and the reaction time is 60-720 min.

Technical Field

The invention relates to the technical field of organic synthesis, in particular to a synthetic method of a dihydrothiazole heterocyclic compound and application of the dihydrothiazole heterocyclic compound in modification of biomolecules.

Background

Dihydrothiazole heterocycles are common in biologically relevant small molecules, such as firefly Luciferin (Luciferin), Desferrithiocin, Micacocidin, and the like (see SYNTHESIS 2013,45, 2763). The structure has good biocompatibility, and has functions of participating in biological reaction and regulating life process due to the unique heterocyclic structure, and has unique medicinal value (see J.Am.chem.Soc.1963,85,337). In addition, by further reaction, a thiazole structure can be obtained using dihydrothiazole to synthesize a drug having a thiazole heterocycle (see j.med. chem.2011,54,4678).

The synthesis of dihydrothiazole heterocycles usually requires long and vigorous conditions. The dihydrothiazole heterocyclic structure can be obtained by reacting cyano with cysteine in water (see J.Med.chem.1999,42,2432), and the reaction time is usually 3-7 days under the condition. Maltsev et al (see SYNTHEESIS 2013,45, 2763-2767) further optimize the reaction conditions, and the reaction can be completed within 12-24 h by using a methanol reflux method. With the above methods, many biologically active structures are gradually inactivated in the reaction. This makes the above method only useful in the synthesis of small molecule drugs, and it is difficult to combine the dihydrothiazole heterocycle with the existing bioactive structure to further develop an improved compound having the dihydrothiazole heterocycle on the basis of the existing bioactive structure.

Among the existing methods, the method capable of obtaining the dihydrothiazole heterocyclic structure mildly and efficiently is the reaction of 2-Cyanobenzothiazole (CBT) with cysteine (see angelw Chem Int Ed engl.2009,48(51),9658), which can occur under mild physiological conditions, and which is fast and efficient, and which can still complete the reaction within about 1h when the reactant concentration is micromole per liter. In addition, the reaction has good specificity and is not easy to interfere by other substances in a receptor system. The characteristics of the reaction can be used as a method for obtaining the dihydrothiazole heterocyclic structure, and can also be used as an efficient reaction to be applied to the marking and modification of various biological systems.

Biological systems are the focus of current chemical and life science research. The labeling and modification of biomolecules are of great significance to research and application, and are important tools for exploring natural systems, creating therapeutic agents and creating new biomolecules. In order to label or modify biomolecules quickly, efficiently and gently, a series of reactions have been developed. These reactions generally have the following characteristics: has good chemoselectivity and regioselectivity, and can be rapidly carried out in aqueous solution at normal temperature (37 ℃, pH6-8 aqueous solution). The aforementioned reaction of CBT with cysteine also belongs to the class of reactions, which can specifically modify the cysteine at the nitrogen terminus of the protein. In addition, azide-alkyne cycloaddition reaction, Staudinger reaction and the like are also commonly used reactions (see Angew. chem. int. Ed.41,2596, Science 287,2007). These existing modifications to natural or unnatural amino acids have formed a more sophisticated and systematic tool kit. Compared to simply modifying biomolecules, it is expected that new methods may offer many advantages over existing tools, such as better simulation of existing structures in organisms, faster modification rates, in vivo reactivity, and more environmental friendliness (see Nature Comm.5, 4740).

Disclosure of Invention

The invention aims to solve the technical problems that the existing reaction conditions for preparing the dihydrothiazole heterocyclic compound are harsh, an efficient and natural modification method is lacked, and the like, and provides a synthesis method of the dihydrothiazole heterocyclic compound and application of the dihydrothiazole heterocyclic compound in modification of biological molecules.

In order to achieve the purpose, the invention adopts the following technical scheme:

a synthetic method of a dihydrothiazole heterocyclic compound comprises the steps of mixing a thioether compound and a compound containing a 1, 2-mercaptoethylamine framework structure in a solvent, adding weak base for reaction, and extracting and rotary steaming to obtain the dihydrothiazole heterocyclic compound; the reaction temperature is 20-60 ℃, and the reaction time is 60-240 min.

According to the synthesis method of the dihydrothiazole heterocyclic compound, the structure general formula of the thioether compound is as follows:

wherein X is sulfur or oxygen, R₁Is an alkyl chain, alkoxy chain, cycloalkyl or aryl, optionally substituted or unsubstituted, except for a mercapto group, amino substituent; r₂Is phenyl optionally substituted or unsubstituted except for mercapto, amino substituents;

the compound containing the 1, 2-mercaptoethylamine framework structure has the following structural general formula:

wherein R is₃Is hydrogen or an optionally substituted group.

The molar ratio of the thiolene ether compound to the compound containing the 1, 2-mercaptoethylamine skeleton structure is 1 (1-2). Preferably, the molar ratio of the thioether compound to the compound containing the 1, 2-mercaptoethylamine skeleton structure is 1: 1.

The solvent comprises at least one of water, methanol, ethanol and acetonitrile; the weak base comprises at least one of sodium carbonate, sodium bicarbonate, potassium carbonate, phosphate buffer and triethylamine.

Preferably, the solvent is a mixed solvent of water and acetonitrile, and the volume ratio is 1: 1; the weak base is sodium bicarbonate.

A synthetic method of a dihydrothiazole heterocyclic compound is applied to modification of biological molecules, and the modified biological molecules connected by a dihydrothiazole heterocyclic skeleton can be obtained by mixing and reacting a thioether compound and the biological molecules containing cysteine residues at the nitrogen end, such as polypeptide or protein, in a weak alkali environment.

The synthesis method of the dihydrothiazole heterocyclic compound is applied to modification of biological molecules, and the structural general formula of the thioether compound is as follows:

wherein X is sulfur or oxygen, R₁Is an alkyl chain, alkoxy chain, cycloalkyl or aryl, optionally substituted or unsubstituted, except for a mercapto group, amino substituent; r₂Is phenyl optionally substituted or unsubstituted except for mercapto, amino substituents; y is covalently bonded to R₂Linked dyes, drugs, functional biomolecules; the molar ratio of the thioether compound to the biomolecule to be modified is (1-100): 1.

The synthesis method of the dihydrothiazole heterocyclic compound is applied to modification of biomolecules, preferably, the weak base is a phosphoric acid buffer solution, the pH value is 7.4, the reaction temperature is 25-37 ℃, and the reaction time is 60-720 min.

Compared with the prior art, the technical scheme of the invention has the following beneficial effects:

the synthesis method of the dihydrothiazole heterocyclic compound is mild and efficient, can quickly react at room temperature and in water, and overcomes the defects of violent reaction conditions, long time and the like in the traditional synthesis method; the method has the advantages of green and economic raw materials, no metal catalyst, environmental friendliness and accordance with the concept of green chemistry. Based on the synthesis method, the invention provides a mild and efficient biomolecule modification method, and biomolecules such as polypeptides and proteins containing a nitrogen-terminal cysteine structure can be efficiently and accurately modified by using the modification method to obtain labeled or functionalized biomolecules so as to further develop applications such as detection and treatment.

Drawings

FIG. 1 is a schematic diagram of the reaction and modification process of the present invention;

FIG. 2 is a schematic diagram of the present invention applied to a biomolecule modification method;

FIG. 3 is a high performance liquid chromatography characterization chart of the biomolecule modification method.

Detailed Description

In order to make the technical problems, technical solutions and advantageous effects of the present invention clearer and clearer, the present invention is further described in detail below with reference to the accompanying drawings and embodiments.

As shown in figure 1, organic or biological molecules with a 1, 2-mercaptoethylamine skeleton structure and a thioether compound are mixed in a weakly alkaline aqueous environment and react for a period of time at room temperature to obtain the dihydrothiazole heterocyclic molecules or modified biological molecules.