A method for preparing a key intermediate of peviptide

Abstract

The invention discloses a method for preparing a key intermediate of pevi peptide, and relates to the technical field of drug synthesis. The specific preparation method includes: first, 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide is reacted with 18-hydroxyoctadecanoic acid tert-butyl ester under the action of a catalyst to generate a glycoside intermediate, the glycoside intermediate is hydrolyzed to remove the acetyl protecting group, and finally oxidized and purified to obtain the target product 18-([β-D-glucuronic acid-1-yl]oxy) tert-butyl octadecanoate, i.e., pevi peptide key intermediate. The preparation method of the present invention can ultimately obtain a key intermediate of pevi peptide with a higher yield and purity, and is simple to operate and has good application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of drug synthesis, and in particular to a method for preparing a key intermediate of pegvitide. Background Art

[0002] Pemvidutide (CAS No. 2538014-94-5) is a novel, investigational, peptide-based dual GLP-1 / glucagon receptor agonist being developed for the treatment of obesity and metabolic dysfunction-associated steatohepatitis (MASH), formerly known as non-alcoholic steatohepatitis (NASH). Activation of the GLP-1 and glucagon receptors is believed to mimic the complementary effects of diet and exercise on weight loss, with GLP-1 suppressing appetite and glucagon increasing energy expenditure. Glucagon is also believed to have a direct effect on hepatic fat metabolism, leading to a rapid reduction in liver fat levels.

[0003] Tert-butyl 18-([β-D-glucuronyl]oxy)octadecanoate is a key starting material in the synthesis of pevitide. Currently, there are few reports on the synthesis of this intermediate. Therefore, a method for effectively preparing this intermediate is still needed. The preparation of this intermediate focuses on controlling the β-configuration. Using 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide as a starting material and a catalyst, a β-configured glycoside intermediate can be obtained. This glycoside intermediate can be used to efficiently synthesize tert-butyl 18-([β-D-glucuronyl]oxy)octadecanoate. Summary of the Invention

[0004] The object of the present invention is to provide a preparation method of a key intermediate of pevi peptide, which can efficiently synthesize 18-([β-D-glucuronic acid-1-yl]oxy) tert-butyl octadecanoate and improve the yield and purity of the product.

[0005] In order to solve the above technical problems, the present invention specifically provides the following technical solutions:

[0006] The present invention discloses a method for preparing a key intermediate of peviptide, comprising:

[0007] S1: mixing tert-butyl 18-hydroxyoctadecanoate and a solvent, and reacting them with 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide in the presence of a catalyst under nitrogen to form a glycoside intermediate; the molar ratio of the tert-butyl 18-hydroxyoctadecanoate to the 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide is 1:0.8-1.2;

[0008] S2: mixing the glycoside intermediate prepared in S1 with a solvent, and then hydrolyzing the mixture with alkaline solution to obtain an oily substance;

[0009] S3: Add a solvent to the oily substance prepared in S2, and oxidize it under the action of an oxidant system to generate a crude product. The crude product is purified by column chromatography to obtain tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate, which is a key intermediate of peviptide.

[0010] Preferably, the solvent in S1 is toluene, and the ratio of tert-butyl 18-hydroxyoctadecanoate to the solvent is 1 mol:4-6 L.

[0011] Preferably, the catalyst in S1 is a metal salt, a Lewis acid or a phase transfer agent, and the molar ratio of tert-butyl 18-hydroxyoctadecanoate to the catalyst is 1:1-6.

[0012] Preferably, the solvent in S2 is methanol, and the ratio of tert-butyl 18-hydroxyoctadecanoate used in preparing the glycoside intermediate to the solvent used in preparing the oil is 1 mol:4-6 L.

[0013] Preferably, the alkali solution in S2 is at least one of sodium methoxide solution, lithium hydroxide solution, and sodium hydroxide solution, and the pH is adjusted to 9-11 with alkali.

[0014] Preferably, the solvent in S3 is at least one of 1,4-dioxane, tetrahydrofuran and methyl tert-butyl ether, and the ratio of tert-butyl 18-hydroxyoctadecanoate in preparing the glycoside intermediate to the solvent in preparing the crude product is 1 mol: 4-6 L.

[0015] Preferably, the oxidant system in S3 is at least one of 2,2,6,6-tetramethylpiperidinium oxide / potassium bromide / sodium hypochlorite solution, Jones reagent and Collins reagent; the molar ratio of tert-butyl 18-hydroxyoctadecanoate used in preparing the glycoside intermediate to 2,2,6,6-tetramethylpiperidinium oxide used in preparing the crude product is 1:0.005-0.02, the molar ratio of 2,2,6,6-tetramethylpiperidinium oxide used in preparing the crude product is 1:8-12, and the effective chlorine content of the sodium hypochlorite solution is 6-14%; the ratio of tert-butyl 18-hydroxyoctadecanoate used in preparing the glycoside intermediate to Jones reagent or Collins reagent used in preparing the crude product is 1 mol:0.8-1.2L.

[0016] Preferably, the column used for purification is a modified silica gel column, which is made of modified silica gel, and the modified silica gel is prepared from silica gel, nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane. Silica gel can be bonded and introduced into a variety of functional groups with different properties by reacting with organic matter to obtain modified silica gel. The surface function of the modified silica gel after the introduction of functional groups is richer, and it shows excellent adsorption capacity for the key intermediate of pevi peptide, i.e. 18-([β-D-glucuronic acid-1-yl]oxy) tert-butyl octadecanoate, and has good yield and purity in the process of purifying the crude product, and can achieve efficient separation.

[0017] Preferably, the reaction temperature in S1 is 20-30°C, and the reaction time is 10-15 hours; the reaction temperature in S2 is 20-30°C, and the reaction time is 3-5 hours; the reaction temperature in S3 is 5-15°C, and the reaction time is 3-5 hours.

[0018] The present invention also discloses the application of a key intermediate of Pevdal prepared by any of the above methods in the preparation of Pevdal.

[0019] The present invention discloses a method for preparing a key intermediate of peviptide, comprising:

[0020] S1: 18-hydroxyoctadecanoic acid tert-butyl ester and a solvent are mixed, and under nitrogen, a catalyst and 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide are added. The mixture is reacted at 20-30°C for 10-15 hours. After the reaction is completed, the reaction is quenched, and the organic phase is separated, washed, dried, and concentrated to obtain a glycoside intermediate.

[0021] S2: The glycoside intermediate prepared in S1 is mixed with a solvent, and then an alkali solution is added at 20-30°C to adjust the pH to 9-11, and the reaction is carried out for 3-5 hours. After the reaction is completed, an acid solution is added to adjust the pH to 6.5-7.5, and then an oil is obtained by extraction and concentration;

[0022] S3: Add a solvent to the oil prepared in S2, lower the temperature to 5-15°C, and then add an oxidant system to react for 3-5 hours. After the reaction, adjust the pH to 2.5-3.5, then extract and separate the organic phase and concentrate to obtain a crude product. The crude product is purified by column chromatography, and finally eluted and concentrated to obtain tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate, a key intermediate of peviptide.

[0023] Preferably, the solvent in S1 is toluene, and the ratio of tert-butyl 18-hydroxyoctadecanoate to the solvent is 1 mol:4-6 L.

[0024] Preferably, the catalyst in S1 is a metal salt, a Lewis acid or a phase transfer agent, and the molar ratio of tert-butyl 18-hydroxyoctadecanoate to the catalyst is 1:1-6.

[0025] More preferably, the metal salt is at least one of silver trifluoromethanesulfonate, silver oxide, silver nitrate, silver carbonate, silver perchlorate, mercuric chloride, mercuric iodide, and mercuric bromide.

[0026] More preferably, the Lewis acid is at least one of Sn(OTf)2, SnCl4, and TrCl-ZnCl2.

[0027] More preferably, the phase transfer agent is at least one of tetrabutylammonium bromide, benzyltriethylammonium chloride and benzyltriethylammonium bromide.

[0028] Preferably, the molar ratio of tert-butyl 18-hydroxyoctadecanoate to 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide in S1 is 1:0.8-1.2.

[0029] Preferably, the solvent used for quenching the reaction in S1 is water.

[0030] Preferably, the solvent used for washing in S1 is a sodium chloride solution, which consists of sodium chloride and water, and the usage ratio of sodium chloride to water is 1 mol: 1-1.5 L.

[0031] Preferably, the solvent in S2 is methanol, and the ratio of tert-butyl 18-hydroxyoctadecanoate used in preparing the glycoside intermediate to the solvent used in preparing the oil is 1 mol:4-6 L.

[0032] Preferably, the alkali solution in S2 is at least one of sodium methoxide solution, lithium hydroxide solution and sodium hydroxide solution.

[0033] More preferably, the sodium methoxide solution consists of sodium methoxide and methanol, and the usage ratio of sodium methoxide to methanol is 1 mol:0.8-1.2 L.

[0034] More preferably, the lithium hydroxide solution consists of lithium hydroxide and water, and the usage ratio of lithium hydroxide to water is 1 mol: 0.8-1.2 L.

[0035] More preferably, the sodium hydroxide solution consists of sodium hydroxide and water, and the usage ratio of sodium hydroxide to water is 1 mol:0.8-1.2 L.

[0036] Preferably, the acid solution in S2 is a hydrochloric acid solution, which consists of hydrochloric acid and water, and the usage ratio of hydrochloric acid to water is 1 mol: 0.8-1.2 L.

[0037] Preferably, the solvent used for extraction in S2 is dichloromethane.

[0038] Preferably, the solvent in S3 is at least one of 1,4-dioxane, tetrahydrofuran and methyl tert-butyl ether, and the ratio of tert-butyl 18-hydroxyoctadecanoate in preparing the glycoside intermediate to the solvent in preparing the crude product is 1 mol: 4-6 L.

[0039] Preferably, the oxidant system in S3 is at least one of 2,2,6,6-tetramethylpiperidinium oxide / potassium bromide / sodium hypochlorite solution, Jones reagent and Collins reagent.

[0040] More preferably, if the oxidant system is 2,2,6,6-tetramethylpiperidinium oxide / potassium bromide / sodium hypochlorite solution, the molar ratio of tert-butyl 18-hydroxyoctadecanoate used in preparing the glycoside intermediate and 2,2,6,6-tetramethylpiperidinium oxide used in preparing the crude product is 1:0.005-0.02, the molar ratio of 2,2,6,6-tetramethylpiperidinium oxide used to potassium bromide is 1:8-12, and the available chlorine content of the sodium hypochlorite solution is 6-14%.

[0041] More preferably, if the oxidant system is Jones reagent or Collins reagent, the ratio of tert-butyl 18-hydroxyoctadecanoate used in preparing the glycoside intermediate to Jones reagent or Collins reagent used in preparing the crude product is 1 mol: 0.8-1.2 L.

[0042] More preferably, if the oxidant system is Jones reagent or Collins reagent, isopropanol needs to be added after the reaction to quench the reaction.

[0043] Preferably, the solvent used to adjust the pH in S3 is a hydrochloric acid solution or a sodium hydroxide solution. The hydrochloric acid solution consists of hydrochloric acid and water, and the ratio of hydrochloric acid to water is 1 mol: 0.8-1.2 L; the sodium hydroxide solution consists of sodium hydroxide and water, and the ratio of sodium hydroxide to water is 1 mol: 0.8-1.2 L.

[0044] Preferably, the column used for purification is a modified silica gel column, which is made of modified silica gel.

[0045] Preferably, the solvent used for extraction in S3 is ethyl acetate.

[0046] Preferably, the solvent used for elution in S3 is a methanol-dichloromethane solution, which consists of methanol and dichloromethane, and the volume ratio of methanol to dichloromethane is 1:18-20.

[0047] The present invention discloses a method for preparing modified silica gel, which is specifically as follows:

[0048] The silica gel is soaked in a hydrochloric acid solution for 20-30 hours, then washed with water and dried to obtain activated silica gel. Under nitrogen conditions, nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane, 3-mercaptopropylmethyldimethoxysilane and toluene are added to the activated silica gel, and the mixture is reacted at 100-130°C for 5-8 hours. After the reaction is completed, the modified silica gel is washed and dried.

[0049] Preferably, the hydrochloric acid solution consists of hydrochloric acid and water, and the volume ratio of hydrochloric acid to water is 1:0.8-1.2.

[0050] Preferably, the mass ratio of the activated silica gel to the N-(acetylglycyl)-3-aminopropyltrimethoxysilane is 1:0.3-0.7.

[0051] Preferably, the mass ratio of N-(acetylglycyl)-3-aminopropyltrimethoxysilane to 3-mercaptopropylmethyldimethoxysilane is 1:0.8-1.2.

[0052] Preferably, the ratio of activated silica gel to toluene is 1 g: 15-25 ml.

[0053] Preferably, the washing is with toluene and then methanol.

[0054] The present invention discloses a method for preparing a modified silica gel column, which is specifically:

[0055] First, the column is pre-filled with eluent, and then the modified silica gel soaked with the eluent is poured into the column. After the modified silica gel is deposited, nitrogen is pressurized and the eluent flows out to obtain a modified silica gel column.

[0056] Preferably, the eluent is a methanol-dichloromethane solution, which consists of methanol and dichloromethane, and the volume ratio of methanol to dichloromethane is 1:18-20.

[0057] More preferably, in the process of preparing modified silica gel according to the present invention, N-((4-vinylbenzyl)aminoidene)diacetic acid may be further used in addition to N-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane to further prepare the modified silica gel. The introduction of N-((4-vinylbenzyl)aminoidene)diacetic acid can construct a synergistic system with multiple functional groups, further changing the properties of the modified silica gel, improving the performance of the modified silica gel, and facilitating the adsorption of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate, thereby enhancing separation and purification capabilities.

[0058] Preferably, the mass ratio of the modified silica gel intermediate to the amount of N-((4-vinylbenzyl)aminoidene)diacetic acid used is 1:0.3-0.8.

[0059] Compared with the prior art, the present invention has the following beneficial effects:

[0060] The present invention provides a method for preparing a key intermediate of pevopeptide. First, 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide is reacted with 18-hydroxyoctadecanoic acid tert-butyl ester in the presence of a catalyst to generate a glycoside intermediate. The glycoside intermediate is then subjected to alkaline hydrolysis to remove the acetyl protecting group. Finally, the target product, 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoic acid tert-butyl ester, is obtained through oxidation reaction and purification. The preparation method of the present invention can ultimately obtain the key intermediate of pevopeptide with high yield and purity, is simple to operate, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for the embodiments or the description of the prior art. Obviously, the drawings described below are merely exemplary, and those skilled in the art can derive other implementation drawings based on the provided drawings without inventive effort.

[0062] Figure 1 This is a graph showing the yield measurement results of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate. DETAILED DESCRIPTION

[0063] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0064] The following first describes the concepts involved in this application with reference to the accompanying drawings. It should be noted that the following description of each concept is intended only to make the content of this application easier to understand and does not limit the scope of protection of this application. At the same time, the embodiments and features in the embodiments of this application can be combined with each other unless there is a conflict. The following detailed description of this application will be made with reference to the accompanying drawings and in conjunction with the embodiments.

[0065] The abbreviations used in the specification and claims have the following meanings:

[0066] Example 1:

[0067] Preparation of modified silica gel: Silica gel was soaked in a hydrochloric acid solution for 24 hours, then washed with water and dried to obtain activated silica gel. Under nitrogen conditions, nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane, 3-mercaptopropylmethyldimethoxysilane, and toluene were added to the activated silica gel and reacted at 120°C for 6 hours. After the reaction, the silica gel was washed with toluene and methanol in sequence and finally dried to obtain the modified silica gel. The hydrochloric acid solution consisted of hydrochloric acid and water, with a volume ratio of hydrochloric acid to water of 1:1. The mass ratio of activated silica gel to nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane was 1:0.5, the mass ratio of nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane to 3-mercaptopropylmethyldimethoxysilane was 1:1, and the ratio of activated silica gel to toluene was 1g:20ml.

[0068] Preparation of a modified silica gel column: First, prefill the column with eluent, then pour the modified silica gel soaked in the eluent into the column. After the modified silica gel is deposited, nitrogen is applied to pressurize the column to dislodge any remaining bubbles, compact the bed, and allow the eluent to flow out to form the modified silica gel column. The eluent is a methanol-dichloromethane solution, which consists of methanol and dichloromethane in a volume ratio of 1:19.

[0069] Preparation of the glycoside intermediate: 18-hydroxyoctadecanoic acid tert-butyl ester and toluene were mixed, and then silver trifluoromethanesulfonate and 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide were added under nitrogen. The mixture was stirred at 25°C for 12 hours. After completion of the reaction, water was added to quench the reaction. The organic phase was separated, washed with sodium chloride solution, dried, and concentrated to obtain the glycoside intermediate. The molar ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to toluene was 1 mol:5 L, the molar ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to silver trifluoromethanesulfonate was 1:2, and the molar ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide was 1:1. The sodium chloride solution consisted of sodium chloride and water, and the molar ratio of sodium chloride to water was 1 mol:1.12 L.

[0070] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The prepared glycoside intermediate was mixed with methanol, and then sodium methoxide solution was added dropwise at 25°C to adjust the pH to 9 for 4 hours. The pH was rechecked every hour during the reaction and maintained at 9. After the reaction, hydrochloric acid solution was added to adjust the pH to 7, and then dichloromethane was added to extract and separate the organic phase. The organic phase was concentrated to obtain an oil. 1,4-Dioxane was added to the oil, the temperature was lowered to 5°C, and 2,2,6,6-tetramethylpiperidinium oxide and potassium bromide were added. Sodium hypochlorite solution was then added dropwise to allow the reaction to proceed for 4 hours. After the reaction, hydrochloric acid solution was added to adjust the pH to 3, and the organic phase was extracted and separated with ethyl acetate. The organic phase was concentrated to obtain the crude product. The crude product was purified by modified silica gel column chromatography, eluted with methanol and dichloromethane, and the eluate was collected. The eluate was concentrated to obtain tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate. The ratio of tert-butyl 18-hydroxyoctadecanoate in the preparation of the glycoside intermediate to methanol in the preparation of the oil is 1 mol:5 L, the sodium methoxide solution is composed of sodium methoxide and methanol, and the ratio of sodium methoxide to methanol is 1 mol:1 L, the hydrochloric acid solution is composed of hydrochloric acid and water, and the ratio of hydrochloric acid to water is 1 mol:1 L, the ratio of tert-butyl 18-hydroxyoctadecanoate in the preparation of the glycoside intermediate to 1,4-dioxane in the preparation of the crude product is 1 mol:5 L, the molar ratio of tert-butyl 18-hydroxyoctadecanoate in the preparation of the glycoside intermediate to 2,2,6,6-tetramethylpiperidinyl oxide in the preparation of the crude product is 1:0.01, the molar ratio of 2,2,6,6-tetramethylpiperidinyl oxide to potassium bromide is 1:10, the effective chlorine content of the sodium hypochlorite solution is 11%, the methanol-dichloromethane solution is composed of methanol and dichloromethane, and the volume ratio of methanol and dichloromethane is 1:19.

[0071] Example 2:

[0072] The preparation of modified silica gel is the same as in Example 1.

[0073] The preparation of the modified silica gel column was the same as in Example 1.

[0074] Preparation of glycoside intermediates: The preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate in this example was compared with that in Example 1, except that silver trifluoromethanesulfonate was replaced with benzyltriethylammonium bromide. Other conditions and parameters were the same as in Example 1.

[0075] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate in this example is compared with that in Example 1, except that the glycoside intermediate is the glycoside intermediate prepared in this example, and the other conditions and parameters are the same as in Example 1.

[0076] Example 3:

[0077] The preparation of modified silica gel is the same as in Example 1.

[0078] The preparation of the modified silica gel column was the same as in Example 1.

[0079] Preparation of the glycoside intermediate: 18-hydroxyoctadecanoic acid tert-butyl ester and toluene were mixed, and then Sn(OTf)2 and 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide were added under nitrogen. The mixture was stirred at 25°C for 12 hours. After completion of the reaction, water was added to quench the reaction. The organic phase was separated, washed with sodium chloride solution, dried, and concentrated to obtain the glycoside intermediate. The molar ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to toluene was 1 mol:5 L, the molar ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to Sn(OTf)2 was 1:1.5, and the molar ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide was 1:1. The sodium chloride solution consisted of sodium chloride and water, and the molar ratio of sodium chloride to water was 1 mol:1.12 L.

[0080] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate in this example is compared with that in Example 1, except that the glycoside intermediate is the glycoside intermediate prepared in this example, and the other conditions and parameters are the same as in Example 1.

[0081] Example 4:

[0082] The preparation of modified silica gel is the same as in Example 1.

[0083] The preparation of the modified silica gel column was the same as in Example 1.

[0084] Preparation of the glycoside intermediate: 18-hydroxyoctadecanoic acid tert-butyl ester and toluene were mixed, and then, under nitrogen, mercuric chloride and 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide were added. The mixture was stirred at 25°C for 12 hours. After completion of the reaction, water was added to quench the reaction. The organic phase was separated, washed with sodium chloride solution, dried, and concentrated to obtain the glycoside intermediate. The molar ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to toluene was 1 mol:5 L, the molar ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to mercuric chloride was 1:1, and the molar ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide was 1:1. The sodium chloride solution consisted of sodium chloride and water, and the molar ratio of sodium chloride to water was 1 mol:1.12 L.

[0085] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate in this example is compared with that in Example 1, except that the glycoside intermediate is the glycoside intermediate prepared in this example, and the other conditions and parameters are the same as in Example 1.

[0086] Example 5:

[0087] The preparation of modified silica gel is the same as in Example 1.

[0088] The preparation of the modified silica gel column was the same as in Example 1.

[0089] The preparation of the glycoside intermediate was the same as in Example 1.

[0090] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The glycoside intermediate was mixed with methanol, and sodium methoxide solution was added dropwise at 25°C to adjust the pH to 9 for 4 hours. The pH was rechecked every hour during the reaction and maintained at 9. After the reaction, hydrochloric acid solution was added to adjust the pH to 7, and then dichloromethane was added to extract and separate the organic phase. The organic phase was concentrated to obtain an oil. 1,4-dioxane was added to the oil, the temperature was lowered to 5°C, and Jones reagent was added to react for 4 hours. After the reaction, isopropanol was added to quench the reaction, and sodium hydroxide solution was added to adjust the pH to 3. The organic phase was extracted with ethyl acetate and concentrated to obtain the crude product. The crude product was purified by modified silica gel column chromatography, eluting with methanol and dichloromethane solution, collecting the eluate, and concentrating the eluate to obtain tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate. The ratio of tert-butyl 18-hydroxyoctadecanoate in the preparation of the glycoside intermediate to methanol in the preparation of the oil is 1 mol:5 L, the sodium methoxide solution is composed of sodium methoxide and methanol, and the ratio of sodium methoxide to methanol is 1 mol:1 L, the hydrochloric acid solution is composed of hydrochloric acid and water, and the ratio of hydrochloric acid to water is 1 mol:1 L, the ratio of tert-butyl 18-hydroxyoctadecanoate in the preparation of the glycoside intermediate to 1,4-dioxane in the preparation of the crude product is 1 mol:5 L, the sodium hydroxide solution is composed of sodium hydroxide and water, and the ratio of sodium hydroxide to water is 1 mol:1 L, the ratio of tert-butyl 18-hydroxyoctadecanoate in the preparation of the glycoside intermediate to Jones reagent in the preparation of the crude product is 1 mol:1 L, and the methanol-dichloromethane solution is composed of methanol and dichloromethane, and the volume ratio of methanol and dichloromethane is 1:19.

[0091] Example 6:

[0092] The preparation of modified silica gel is the same as in Example 1.

[0093] The preparation of the modified silica gel column was the same as in Example 1.

[0094] The preparation of the glycoside intermediate was the same as in Example 1.

[0095] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate in this example is compared with that in Example 5, except that the Jones reagent is replaced by the Collins reagent, and the other conditions and parameters are the same as in Example 5.

[0096] Example 7:

[0097] Preparation of modified silica gel: The preparation of the modified silica gel column in this embodiment is compared with that in Example 1, except that the mass ratio of activated silica gel to nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane is 1:0.8, and other conditions and parameters are the same as in Example 1.

[0098] Preparation of modified silica gel column: The preparation of the modified silica gel column in this example is compared with that in Example 1, except that the modified silica gel is replaced by the modified silica gel prepared in this example, and the other conditions and parameters are the same as in Example 1.

[0099] The preparation of the glycoside intermediate was the same as in Example 1.

[0100] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate in this example is compared with that in Example 1, except that the modified silica gel column is the modified silica gel column prepared in this example, and the other conditions and parameters are the same as in Example 1.

[0101] Example 8:

[0102] Preparation of modified silica gel: Soak the silica gel in a hydrochloric acid solution for 24 hours, then wash with water and dry to obtain activated silica gel. Under nitrogen conditions, add nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane, 3-mercaptopropylmethyldimethoxysilane and toluene to the activated silica gel, react at 120°C for 6 hours, wash with toluene and methanol after the reaction, and then dry to obtain a modified silica gel intermediate. Add acetonitrile, azobisisobutyronitrile and N-((4-vinylbenzyl)amino)diacetic acid to the modified silica gel intermediate, react at 80°C for 36 hours, wash with acetonitrile and diethyl ether after the reaction, and finally dry to obtain modified silica gel. The hydrochloric acid solution is composed of hydrochloric acid and water, the volume ratio of hydrochloric acid to water is 1:1, the mass ratio of activated silica gel to nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane is 1:1, the mass ratio of nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane to 3-mercaptopropylmethyldimethoxysilane is 1:1, the ratio of activated silica gel to toluene is 1 g:20 ml, the ratio of modified silica gel intermediate to acetonitrile is 1 g:15 ml, the mass ratio of modified silica gel intermediate to azobisisobutyronitrile is 1:0.008, and the mass ratio of modified silica gel intermediate to N-((4-vinylbenzyl)amino)diacetic acid is 1:0.38.

[0103] Preparation of modified silica gel column: The preparation of the modified silica gel column in this example is compared with that in Example 1, except that the modified silica gel is replaced by the modified silica gel prepared in this example, and the other conditions and parameters are the same as in Example 1.

[0104] The preparation of the glycoside intermediate was the same as in Example 1.

[0105] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate in this example is compared with that in Example 1, except that the modified silica gel column is the modified silica gel column prepared in this example, and the other conditions and parameters are the same as in Example 1.

[0106] Example 9:

[0107] Preparation of modified silica gel: The preparation of the modified silica gel column in this example is compared with that in Example 8, except that the mass ratio of the modified silica gel intermediate to N-((4-vinylbenzyl)amino)diacetic acid used is 1:0.6, and other conditions and parameters are the same as in Example 8.

[0108] Preparation of modified silica gel column: The preparation of the modified silica gel column in this example is compared with that in Example 8, except that the modified silica gel is replaced by the modified silica gel prepared in this example, and the other conditions and parameters are the same as in Example 8.

[0109] The preparation of the glycoside intermediate is the same as in Example 8.

[0110] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate in this example is compared with that in Example 8, except that the modified silica gel column is the modified silica gel column prepared in this example, and the other conditions and parameters are the same as in Example 8.

[0111] Comparative Example 1:

[0112] Preparation of modified silica gel: The preparation of the modified silica gel column in this comparative example is compared with that in Example 1, except that the mass ratio of activated silica gel to nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane is 1:0.05, and other conditions and parameters are the same as in Example 1.

[0113] Preparation of modified silica gel column: The preparation of the modified silica gel column in this comparative example is compared with that in Example 1, except that the modified silica gel is replaced by the modified silica gel prepared in this example, and the other conditions and parameters are the same as in Example 1.

[0114] The preparation of the glycoside intermediate was the same as in Example 1.

[0115] Preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate: The preparation of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate in this comparative example is compared with that in Example 1, except that the modified silica gel column is the modified silica gel column prepared in this example, and the other conditions and parameters are the same as in Example 1.

[0116] In the experimental examples of the present invention, HPLC method was used to detect tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate.

[0117] The HPLC method is as follows:

[0118] Reference solution: Accurately weigh 20 mg of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate reference substance into a 10 mL volumetric flask. Dissolve with a dissolving agent and dilute to a volume of 2 mg per mL. This serves as the reference solution. The diluent is acetonitrile-water solution, which is a mixture of acetonitrile and water in a 1:1 volume ratio.

[0119] Test solution: Accurately weigh 20 mg of the test sample into a 10 mL volumetric flask. Dissolve the sample in a diluent and dilute to a volume of 2 mg per mL. This serves as the test solution. The test sample is tert-butyl 18-([β-D-glucuronyl]oxy)octadecanoate prepared in any of the Examples or Comparative Examples. The diluent is an acetonitrile-water solution, prepared by mixing acetonitrile and water in a 1:1 volume ratio.

[0120] Mobile phase: Use phosphoric acid aqueous solution as mobile phase A and acetonitrile as mobile phase B. To prepare the phosphoric acid aqueous solution, accurately measure 1 ml of phosphoric acid into a 1000 ml volumetric flask, add water to the volume and shake well, then filter through a 0.45 μm microporous membrane.

[0121] Detection Method: An Agilent ZORBAX SB-C18 5μm, 4.6mm×250mm column was used with a detection wavelength of 210nm. Gradient elution was performed with aqueous phosphoric acid as mobile phase A and acetonitrile as mobile phase B. The gradient elution program was as follows: at 0 minutes, mobile phase A: 40 vol%, mobile phase B: 60 vol%; at 35 minutes, mobile phase A: 10 vol%, mobile phase B: 90 vol%; at 50 minutes, mobile phase A: 10 vol%, mobile phase B: 90 vol%; at 51 minutes, mobile phase A: 40 vol%, mobile phase B: 60 vol%; at 60 minutes, mobile phase A: 40 vol%, mobile phase B: 60 vol%; the column temperature was 30°C, the flow rate was 1.0 ml / min, and the sample injection volume was 20 μL. The resolution between the main peak and each impurity should be no less than 1.5.

[0122] Experimental Example 1:

[0123] Yield determination: Statistical calculation was performed on the tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate prepared in Examples 1-10 and Comparative Example 1 to obtain the yield.

[0124] The yield of 18-([β-D-glucuronic acid-1-yl]oxy) tert-butyl octadecanoate prepared in Examples 1-10 and Comparative Example 1 was determined as follows: Figure 1As shown, Example 1 is compared with Examples 2-4, indicating that when silver trifluoromethanesulfonate is used as a catalyst, the yield of 18-([β-D-glucuronic acid-1-yl]oxy) octadecanoic acid tert-butyl ester is the highest; Example 1 is compared with Examples 5-6, indicating that when the oxidant system is 2,2,6,6-tetramethylpiperidinyl oxide, potassium bromide and sodium hypochlorite, the yield of 18-([β-D-glucuronic acid-1-yl]oxy) octadecanoic acid tert-butyl ester is the highest; Example 1 is compared with Example 7, indicating that the increase in the usage of nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane within a certain range can improve the effect of modified silica gel, thereby improving the yield of 18-([β-D-glucuronic acid-1-yl]oxy) octadecanoic acid tert-butyl ester; Example 7 is compared with Example 8, indicating that when nitrogen-(acetylglycyl)- On the basis of 3-aminopropyl trimethoxysilane and 3-mercaptopropyl methyl dimethoxysilane, N-((4-vinylbenzyl) aminoidene) diacetic acid is used to prepare modified silica gel, which can further improve the yield of 18-([β-D-glucuronic acid-1-yl] oxygen base) t-butyl octadecanoate; Example 8 is compared with Example 9, and shows that the increase of N-((4-vinylbenzyl) aminoidene) diacetic acid usage amount can also improve the yield of 18-([β-D-glucuronic acid-1-yl] oxygen base) t-butyl octadecanoate prepared subsequently; Example 1 is compared with Comparative Example 1, and shows that the usage amount of nitrogen-(acetylglycyl)-3-aminopropyl trimethoxysilane needs to be in suitable scope, and usage amount is too low to promoting the yield of 18-([β-D-glucuronic acid-1-yl] oxygen base) t-butyl octadecanoate without obvious effect.

[0125] Experimental Example 2:

[0126] Purity determination: The tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate prepared in Examples 1-10 and Comparative Example 1 was subjected to HPLC analysis to obtain the purity.

[0127] Table 1 Purity determination results

[0128]

[0129] The purity determination results of 18-([β-D-glucuronic acid-1-yl] oxygen base) t-butyl octadecanoate prepared in Examples 1-10 and Comparative Example 1 are shown in Table 1. Example 1, compared with Examples 2-4, shows that when silver trifluoromethanesulfonate is used as a catalyst, the purity of 18-([β-D-glucuronic acid-1-yl] oxygen base) t-butyl octadecanoate is the highest; Example 1, compared with Examples 5-6, shows that when the oxidant system is 2,2,6,6-tetramethylpiperidinium oxide, potassium bromide and sodium hypochlorite, the purity of 18-([β-D-glucuronic acid-1-yl] oxygen base) t-butyl octadecanoate is the highest; Example 1, compared with Example 7, shows that the increase in the usage of nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane within a certain range can improve the effect of modified silica gel, thereby improving the purity of 18-([β-D-glucuronic acid-1-yl] oxygen base) t-butyl octadecanoate Embodiment 7 is compared with Example 8, and shows that on the basis of using nitrogen-(acetylglycyl)-3-aminopropyl trimethoxysilane and 3-mercaptopropyl methyl dimethoxy silane, re-use N-((4-vinylbenzyl) aminoidene) diacetic acid to prepare modified silica gel, can further improve the purity of 18-([β-D-glucuronic acid-1-yl] oxygen base) octadecanoic acid tert-butyl ester; Embodiment 8 is compared with Example 9, and shows that the increase of N-((4-vinylbenzyl) aminoidene) diacetic acid usage amount can also improve the purity of 18-([β-D-glucuronic acid-1-yl] oxygen base) octadecanoic acid tert-butyl ester of follow-up preparation; Embodiment 1 is compared with Comparative Example 1, and shows that the usage amount of nitrogen-(acetylglycyl)-3-aminopropyl trimethoxysilane needs to be in suitable scope, and usage amount is too low to the purity of promoting 18-([β-D-glucuronic acid-1-yl] oxygen base) octadecanoic acid tert-butyl ester does not have obvious effect.

[0130] Experimental Example 3:

[0131] To determine the adsorption rate of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate on modified silica gel, add a solution of tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate to the activated silica gel. The mixture was shaken at 150 rpm at 30°C for uniform adsorption. The tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate content in the solution was determined by HPLC, and the time required for adsorption to reach equilibrium was determined. Adsorption rate = (tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate concentration in the solution before adsorption - tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate concentration in the solution after adsorption) / tert-butyl 18-([β-D-glucuronic acid-1-yl]oxy)octadecanoate concentration in the solution before adsorption × 100%.

[0132] Table 2 Determination results of adsorption rate

[0133]

[0134] The results are shown in Table 2. Compared with Example 7, Example 1 shows that increasing the usage of nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane within a certain range can improve the adsorption of modified silica gel; Compared with Example 8, Example 7 shows that using nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane and then using N-((4-vinylbenzyl)aminoidene)diacetic acid to prepare modified silica gel can further improve the adsorption of the prepared modified silica gel; Compared with Example 9, Example 8 shows that increasing the usage of N-((4-vinylbenzyl)aminoidene)diacetic acid can also improve the adsorption of modified silica gel; Compared with Comparative Example 1, Example 1 shows that the usage of nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane needs to be within an appropriate range, and too low an usage amount has no obvious effect on improving the adsorption rate of modified silica gel.

[0135] The embodiments and / or implementation methods described above are only used to illustrate the preferred embodiments and / or implementation methods for realizing the technology of the present invention, and do not impose any form of limitation on the implementation methods of the technology of the present invention. Any person skilled in the art may make slight changes or modifications to other equivalent embodiments without departing from the scope of the technical means disclosed in the content of the present invention, but they should still be regarded as technologies or embodiments that are essentially the same as the present invention.

[0136] This article uses specific examples to illustrate the principles and implementation methods of this application. The description of the above embodiments is only used to help understand the method and core ideas of this application. The above is only the preferred implementation method of this application. It should be pointed out that due to the limitations of textual expression, there are objectively infinite specific structures. For ordinary technicians in this technical field, without departing from the principles of this application, they can also make several improvements, modifications or changes, and can also combine the above technical features in an appropriate manner; these improvements, modifications, changes or combinations, or the direct application of the inventive concept and technical solution to other occasions without improvement, should be regarded as the scope of protection of this application.

Claims

1. A method for preparing a key intermediate of peviptide, comprising: S1: mixing tert-butyl 18-hydroxyoctadecanoate and a solvent, and reacting them with 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide in the presence of a catalyst under nitrogen conditions to form a glycoside intermediate; the molar ratio of the tert-butyl 18-hydroxyoctadecanoate to the 2,3,4,6-tetraacetoxy-α-D-pyranose glucopyranose bromide is 1:0.8-1.2; the catalyst in S1 is benzyltriethylammonium bromide, and the molar ratio of the tert-butyl 18-hydroxyoctadecanoate to the catalyst is 1:1-6; S2: mixing the glycoside intermediate prepared in S1 with a solvent, and then hydrolyzing the mixture with alkaline solution to obtain an oily substance; S3: Add a solvent to the oily substance prepared in S2, oxidize it under the action of an oxidant system to generate a crude product, and purify the crude product by column chromatography to obtain 18-([β-D-glucuronic acid-1-yl]oxy) octadecanoic acid tert-butyl ester, which is the key intermediate of pevi peptide; the solvent in S3 is at least one of 1,4-dioxane, tetrahydrofuran and methyl tert-butyl ether, and the ratio of 18-hydroxyoctadecanoic acid tert-butyl ester to the solvent used in preparing the crude product when preparing the glycoside intermediate is 1 mol: 4-6L. The column used for purification in S3 is a modified silica gel column, which is made of modified silica gel, and the modified silica gel is prepared from silica gel, nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane.

2. The method for preparing a key intermediate of Pevitil according to claim 1, wherein: The solvent in S1 is toluene, and the ratio of tert-butyl 18-hydroxyoctadecanoate to the solvent is 1 mol:4-6 L.

3. The method for preparing a key intermediate of Pevitil according to claim 1, wherein: The solvent in S2 is methanol, and the ratio of tert-butyl 18-hydroxyoctadecanoate used in preparing the glycoside intermediate to the solvent used in preparing the oil is 1 mol: 4-6 L.

4. The method for preparing a key intermediate of Pevitil according to claim 1, wherein: The alkali solution in S2 is at least one of sodium methoxide solution, lithium hydroxide solution and sodium hydroxide solution, and the pH is adjusted to 9-11 with alkali.

5. The method for preparing a key intermediate of Pevitil according to claim 1, characterized in that: The oxidant system in S3 is at least one of 2,2,6,6-tetramethylpiperidinium oxide / potassium bromide / sodium hypochlorite solution, Jones reagent, and Collins reagent; the molar ratio of tert-butyl 18-hydroxyoctadecanoate used in preparing the glycoside intermediate to 2,2,6,6-tetramethylpiperidinium oxide used in preparing the crude product is 1:0.005-0.02, the molar ratio of 2,2,6,6-tetramethylpiperidinium oxide used in preparing the crude product is 1:8-12, and the effective chlorine content of the sodium hypochlorite solution is 6-14%; the ratio of tert-butyl 18-hydroxyoctadecanoate used in preparing the glycoside intermediate to Jones reagent or Collins reagent used in preparing the crude product is 1 mol:0.8-1.2 L.

6. The method for preparing a key intermediate of Pevitil according to claim 1, characterized in that: In the S1, the reaction temperature is 20-30°C, and the reaction time is 10-15 hours; in the S2, the reaction temperature is 20-30°C, and the reaction time is 3-5 hours; in the S3, the reaction temperature is 5-15°C, and the reaction time is 3-5 hours.

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