阅读说明：本技术 一种免疫调节剂的关键中间体合成方法 (Method for synthesizing key intermediate of immunomodulator ) 是由 黄保华 刘小栋 马良鑫 胡新礼 戴建 于 2020-04-08 设计创作，主要内容包括：本发明提供了一种免疫调节剂的关键中间体合成方法,该方法通过将化合物2和化合物3溶解,加入碳酸钾,置换氮气并加入S-Phos升反应；结束后降温过滤,浓缩,有机相经洗涤干燥得化合物4；将其溶于四氢呋喃中,加入N,O-二甲基羟胺盐酸盐和三乙胺；室温搅拌,淬灭后加二氯甲烷,用氯化铵水溶液洗涤；有机相经干燥后,用四氢呋喃进行浓缩,在四氢呋喃溶液中加入二异丁基氢化铝反应后,加硫酸氢钾水溶液,得混合物过滤后,加二氯甲烷和盐酸洗涤；有机相浓缩至干得化合物5；将其和化合物6混合二氯甲烷中反应,加入Dess-Martin氧化试剂,纯化得化合物1。本发明路线仅有一步使用了金属催化,且该步骤远离最终化合物,极大降低了药物分子中的金属残留。(The invention provides a method for synthesizing a key intermediate of an immunomodulator, which comprises the steps of dissolving a compound 2 and a compound 3, adding potassium carbonate, replacing nitrogen and adding S-Phos liter for reaction; cooling, filtering, concentrating, washing and drying the organic phase to obtain a compound 4; dissolving the mixture in tetrahydrofuran, and adding N, O-dimethylhydroxylamine hydrochloride and triethylamine; stirring at room temperature, quenching, adding dichloromethane, and washing with ammonium chloride aqueous solution; drying the organic phase, concentrating with tetrahydrofuran, adding diisobutylaluminum hydride into tetrahydrofuran solution, reacting, adding potassium hydrogen sulfate water solution to obtain mixture, filtering, and washing with dichloromethane and hydrochloric acid; concentrating the organic phase to dryness to obtain a compound 5; the mixture reacts with compound 6 in mixed dichloromethane, Dess-Martin oxidation reagent is added, and compound 1 is obtained after purification. The route of the invention uses metal catalysis in only one step, and the step is far away from the final compound, thus greatly reducing the metal residue in the drug molecule.)

1. A method for synthesizing a key intermediate of an immunomodulator is characterized in that: the immunomodulator is a PD-I ligand, the intermediate has a structural formula shown in the specification,

the synthesis reaction is as follows:

；

comprises the following steps of (a) carrying out,

s1, dissolving the compound 2 and the compound 3 in ethanol, adding potassium carbonate, replacing nitrogen for 2-4 times, adding S-Phos under the protection of nitrogen, and heating to 20-50 ℃ for reaction for 10-15 hours;

s2, after the reaction is finished, cooling to room temperature, filtering, concentrating, adding water and dichloromethane, washing an organic phase by using dilute hydrochloric acid, and drying by using anhydrous sodium sulfate to obtain a crude product of the compound 4;

s3, dissolving the compound 4 in tetrahydrofuran, adding N, O-dimethylhydroxylamine hydrochloride and triethylamine, and EDCI; stirring at room temperature, adding water to quench and react, adding dichloromethane, and washing with ammonium chloride aqueous solution; the organic phase was dried over anhydrous sodium sulfate and concentrated with dry tetrahydrofuran.

S4, adding diisobutyl aluminum hydride into the obtained tetrahydrofuran solution at the temperature of between 60 ℃ below zero and 90 ℃ below zero for reaction;

s5, after the reaction is finished, adding a potassium bisulfate aqueous solution, filtering the obtained mixture, and adding dichloromethane and 1N hydrochloric acid for washing; concentrating the obtained organic phase to dryness to obtain a compound 5;

and S6, the compound 5 and the compound 6 are mixed in dry dichloromethane and react for 30 minutes, then a Dess-Martin oxidation reagent is added, and after the detection reaction is finished, the compound 1 is obtained by only silica gel column purification.

2. The method of claim 1, wherein the method comprises the steps of: the S1 was purged with nitrogen 3 times, and 2 mol% of S-Phos was added under nitrogen protection.

3. The method of claim 1, wherein the method comprises the steps of: and the S2 further comprises a purification step of the crude product, wherein the purification step is to purify the crude product by column chromatography to obtain a refined compound 4.

4. A method of synthesizing key intermediates of an immunomodulator according to claim 2, wherein: the mixture was stirred at room temperature for 10 to 15 hours in S3.

5. The method of claim 4, wherein the key intermediate of the immunomodulator comprises: the organic phase in S3 was dried over anhydrous sodium sulfate and concentrated 2-4 times with dry tetrahydrofuran.

6. The method of claim 4, wherein the key intermediate of the immunomodulator comprises: in the S5, after the reaction is finished, the potassium bisulfate aqueous solution is added at a temperature of below 0 ℃.

Technical Field

The invention belongs to the technical field of chemical synthesis, and particularly relates to a method for synthesizing a key intermediate of an immunomodulator.

Background

The immune system plays an important role in controlling and eradicating diseases such as cancer. However, cancer cells can develop new approaches to evade or suppress the immune system to facilitate their growth. One mechanism is to alter the expression of costimulatory and costimulatory molecules on immune cells. Blocking signals from inhibitory immune checkpoints, such as PD-I, has proven to be a promising and effective treatment modality.

Programmed death receptor 1(PD-I), also known as CD279, is a cell surface receptor expressed on activated T cells, natural killer T cells, B cells and macrophages. It acts as an intrinsic negative feedback system preventing activation of T cells, which in turn reduces autoimmunity and promotes self-tolerance. In addition, PD-1 plays a key role in suppressing its response to antigen-specific T cells in diseases such as cancer and viral infection.

Several lines of evidence in preclinical animal studies suggest that PD-I and its ligands negatively regulate immune responses. PD-I deficient mice have been shown to develop lupus-like adrenalitis and dilated cardiomyopathy. Using the LCMV model of chronic infection, it was shown that PD-I/PD-LI interactions inhibit activation, expansion and acquisition of effector functions of virus-specific CD8T cells. Together, these data support the development of a therapeutic approach to prevent the PD-I mediated inhibitory signaling cascade to enhance the enhancer 'rescue' T cell response. Therefore, new compounds are needed to prevent PD-I/PD-LI proteins/proteins from interacting.

Formula 1 is a key intermediate of compounds used to prevent PD-I/PD-LI protein/protein interactions. The intermediate can be derived to obtain a plurality of candidate drugs for PD-I/PD-LI protein/protein interaction.

The existing synthetic method has complicated steps, and the final product possibly has metal substance residues due to multiple metal catalysis in the reaction, so that the purity of the final product is not enough, and the subsequent preparation of the medicine is influenced.

Disclosure of Invention

In order to solve the defects of the prior art, the invention provides a method for synthesizing a key intermediate of an immunomodulator.

The purpose of the invention is realized by the following technical scheme:

a method for synthesizing a key intermediate of an immunomodulator, wherein the immunomodulator is a PD-I ligand, the intermediate has a structural formula shown in the specification,

the synthesis reaction is as follows:

；

comprises the following steps of (a) carrying out,

s1, dissolving the compound 2 and the compound 3 in ethanol, adding potassium carbonate, replacing nitrogen for 2-4 times, adding S-Phos under the protection of nitrogen, and heating to 20-50 ℃ for reaction for 10-15 hours;

s2, after the reaction is finished, cooling to room temperature, filtering, concentrating, adding water and dichloromethane, washing an organic phase by using dilute hydrochloric acid, and drying by using anhydrous sodium sulfate to obtain a crude product of the compound 4;

s3, dissolving the compound 4 in tetrahydrofuran, adding N, O-dimethylhydroxylamine hydrochloride and triethylamine, and EDCI; stirring at room temperature, adding water to quench and react, adding dichloromethane, and washing with ammonium chloride aqueous solution; the organic phase was dried over anhydrous sodium sulfate and concentrated with dry tetrahydrofuran.

S4, adding diisobutyl aluminum hydride into the obtained tetrahydrofuran solution at the temperature of between 60 ℃ below zero and 90 ℃ below zero for reaction;

s5, after the reaction is finished, adding a potassium bisulfate aqueous solution, filtering the obtained mixture, and adding dichloromethane and 1N hydrochloric acid for washing; concentrating the obtained organic phase to dryness to obtain a compound 5;

and S6, the compound 5 and the compound 6 are mixed in dry dichloromethane and react for 30 minutes, then a Dess-Martin oxidation reagent is added, and after the detection reaction is finished, the compound 1 is obtained by only silica gel column purification.

Preferably, the nitrogen in the S1 is replaced 3 times, and 2mol percent of S-Phos is added under the protection of nitrogen.

Preferably, the S2 further comprises a purification step of crude product, wherein the purification step is to purify the crude product by column chromatography to obtain a refined compound 4.

Preferably, the stirring in S3 is performed at room temperature for 10 to 15 hours.

Preferably, the organic phase in S3 is dried over anhydrous sodium sulfate and then concentrated 2-4 times with dried tetrahydrofuran.

Preferably, in the S5, after the reaction is finished, the potassium bisulfate aqueous solution is added below 0 ℃.

The invention has the beneficial effects that: the route of the invention uses metal catalysis in only one step, and the step is far away from the final compound, thus greatly reducing the metal residue in the drug molecule.

Detailed Description

The technical scheme of the invention is specifically illustrated by combining the following embodiments, and the invention discloses a key intermediate synthesis method of an immunomodulator.

Example 1

Preparation of compound 4, the reaction is shown below:

1.2eq of Compound 2 and 340mg of Compound 4 are dissolved in 100mL of ethanol, 5 equivalents of potassium carbonate are added, nitrogen is replaced 3 times, 2 mol% S-Phos is added under nitrogen protection, and the mixture is heated to 40 ℃ for reaction for 12 hours. After the reaction is finished, cooling to room temperature, filtering, concentrating, adding water and dichloromethane, washing an organic phase by using dilute hydrochloric acid, and drying by using anhydrous sodium sulfate. The crude product was purified by column chromatography to give 355mg, 84% yield.

Example 2

The preparation of compound 5, the reaction formula of which is shown below,

400mg of compound 4 are dissolved in tetrahydrofuran and 1.5eq of N, O-dimethylhydroxylamine hydrochloride and 2eq of triethylamine, 1.8eq of EDCI are added. After stirring at room temperature for 12 hours. The reaction was quenched with water, dichloromethane was added, and the mixture was washed with an aqueous ammonium chloride solution. The organic phase was dried over anhydrous sodium sulfate and concentrated 3 times with dry tetrahydrofuran. DIBAL (diisobutylaluminum hydride) was added to the resulting tetrahydrofuran solution at-78 ℃. After the reaction is completed, the potassium bisulfate aqueous solution is added below 0 ℃. The resulting mixture was filtered, and washed with dichloromethane and 1N hydrochloric acid. The organic phase obtained is concentrated to dryness and is directly taken to the next step without purification.

Example 3

Preparation of compound 1, the reaction is shown below:

1.5eq Compound 5 and 167mg Compound 6 were mixed in dry dichloromethane and reacted for 30 minutes, followed by addition of 2eq Dess-Martin oxidation reagent (1,1, 1-triacyloxy-1, 1-dihydro-1, 2-benzidoxol-3 (1H) -one). After the detection reaction was completed, compound 1 was obtained by silica gel column purification alone in 80% yield.

There are, of course, many other specific embodiments of the invention and these are not to be considered as limiting. All technical solutions formed by using equivalent substitutions or equivalent transformations fall within the scope of the claimed invention.