Preparation method of pevitide key intermediate

Abstract

The invention discloses a preparation method of a pevitide key intermediate, and relates to the technical field of medicine synthesis. The specific preparation method comprises the following steps: firstly, reacting 2, 3, 4, 6-tetraacetoxy-alpha-D-glucopyranose bromide with 18-hydroxy tert-butyl octadecanoate under the action of a catalyst to generate a glycoside intermediate, and carrying out alkaline hydrolysis on the glycoside intermediate to remove an acetyl protecting group, so as to obtain the 2, 3, 4, 6-tetraacetoxy-alpha-D-glucopyranose. And finally, carrying out oxidation reaction and purification to obtain a target product 18-([beta-D-glucuronic acid-1-yl] oxy) tert-butyl octadecanoate, namely the pevitide key intermediate. According to the preparation method disclosed by the invention, the pevitide key intermediate with relatively high yield and purity can be finally obtained, and the preparation method is simple to operate and has a good application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of drug synthesis, and particularly relates to a preparation method of a key intermediate of Pemvidutide. Background Art

[0002] Pemvidutide (CAS No.: 2538014-94-5) is a novel, investigational, peptide-based GLP-1 / glucagon dual 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 thought 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 thought to have a direct effect on hepatic lipid metabolism, resulting in a rapid reduction in hepatic lipid levels.

[0003] tert-Butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate is a key starting material in the synthesis of Pemvidutide. Currently, there are few reports on the synthesis of this intermediate. Therefore, there is still a need for a method that can effectively prepare this intermediate. In the preparation process of this intermediate, the control of the β configuration is mainly involved. Using 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide as the raw material, under the action of a catalyst, a glycoside intermediate with the β configuration can be obtained. The glycoside intermediate can be used to effectively synthesize tert-Butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate. Summary of the Invention

[0004] The purpose of the present invention is to provide a preparation method of a key intermediate of Pemvidutide, which can efficiently synthesize tert-Butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate and improve the yield and purity of the product.

[0005] To solve the above technical problems, the present invention specifically provides the following technical solutions: The present invention discloses a preparation method of a key intermediate of Pemvidutide, comprising: S1: Mix tert-Butyl 18-hydroxyoctadecanoate and a solvent, and react with 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide under the action of a catalyst under a nitrogen atmosphere to form a glycoside intermediate; the molar ratio of the amount of tert-Butyl 18-hydroxyoctadecanoate to 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide used is 1:0.8 - 1.2; S2: Mix the glycoside intermediate prepared in S1 with a solvent, and then carry out an alkali hydrolysis reaction to obtain an oil; S3: Add a solvent to the oily substance obtained in S2, and oxidize it under the action of an oxidant system to obtain a crude product. The crude product is purified by column chromatography to obtain tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate, which is the key intermediate of pervipetide.

[0006] Preferably, the solvent in S1 is toluene, and the dosage ratio of tert-butyl 18-hydroxyoctadecanoate to the solvent is 1 mol: 4 - 6 L.

[0007] 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.

[0008] Preferably, the solvent in S2 is methanol, and the dosage ratio of tert-butyl 18-hydroxyoctadecanoate to the solvent when preparing the glycoside intermediate is 1 mol: 4 - 6 L.

[0009] Preferably, the lye 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 the base.

[0010] Preferably, the solvent in S3 is at least one of 1,4-dioxane, tetrahydrofuran and methyl tert-butyl ether, and the dosage ratio of tert-butyl 18-hydroxyoctadecanoate to the solvent when preparing the crude product is 1 mol: 4 - 6 L.

[0011] Preferably, the oxidant system in S3 is at least one of 2,2,6,6-tetramethylpiperidine oxide / potassium bromide / sodium hypochlorite solution, Jones reagent and Collins reagent; the molar ratio of tert-butyl 18-hydroxyoctadecanoate to 2,2,6,6-tetramethylpiperidine oxide when preparing the crude product is 1: 0.005 - 0.02, the molar ratio of 2,2,6,6-tetramethylpiperidine oxide to potassium bromide is 1: 8 - 12, and the effective chlorine content of the sodium hypochlorite solution is 6 - 14%; the dosage ratio of tert-butyl 18-hydroxyoctadecanoate to Jones reagent or Collins reagent when preparing the crude product is 1 mol: 0.8 - 1.2 L.

[0012] Preferably, the column used for purification is a modified silica gel column made of modified silica gel, and the modified silica gel is prepared from silica gel, N-(acetylglycyl)-3-aminopropyltrimethoxysilane, and 3-mercaptopropylmethyldimethoxysilane. By reacting with organic substances, various functional groups with different properties can be bonded and introduced into silica gel to obtain modified silica gel. After introducing functional groups, the surface of the modified silica gel has richer functions, shows excellent adsorption capacity for the key intermediate of pevibepride, namely tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, and has good yield and purity during the purification of crude products, enabling efficient separation.

[0013] Preferably, in S1, the reaction temperature is 20 - 30 °C and the reaction time is 10 - 15 h; in S2, the reaction temperature is 20 - 30 °C and the reaction time is 3 - 5 h; in S3, the reaction temperature is 5 - 15 °C and the reaction time is 3 - 5 h.

[0014] The present invention also discloses the application of the key intermediate of pevibepride prepared by any of the above methods in the preparation of pevibepride.

[0015] The present invention discloses a preparation method of a key intermediate of pevibepride, comprising: S1: Mix tert-butyl 18-hydroxyoctadecanoate and a solvent, add a catalyst and 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide under a nitrogen atmosphere, react at 20 - 30 °C for 10 - 15 h, quench the reaction after the reaction ends, separate the organic phase, and then obtain a glycoside intermediate through washing, drying, and concentration; S2: Mix the glycoside intermediate prepared in S1 with a solvent, then add an alkali solution at 20 - 30 °C to adjust the pH to 9 - 11, react for 3 - 5 h, add an acid solution to adjust the pH to 6.5 - 7.5 after the reaction ends, and then obtain an oil through extraction and concentration; S3: Add a solvent to the oil prepared in S2, lower the temperature to 5 - 15 °C, then add an oxidant system and react for 3 - 5 h, adjust the pH to 2.5 - 3.5 after the reaction ends, 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-glucuronosyl-1-yl]oxy)octadecanoate, namely the key intermediate of pevibepride.

[0016] Preferably, the solvent in S1 is toluene, and the dosage ratio of tert-butyl 18-hydroxyoctadecanoate to the solvent is 1 mol : 4 - 6 L.

[0017] 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 usage amount is 1 : 1 - 6.

[0018] 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.

[0019] More preferably, the Lewis acid is at least one of Sn(OTf)2, SnCl4, and TrCl-ZnCl2.

[0020] More preferably, the phase transfer agent is at least one of tetrabutylammonium bromide, benzyltriethylammonium chloride and benzyltriethylammonium bromide.

[0021] 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.

[0022] Preferably, the solvent used for quenching the reaction in S1 is water.

[0023] 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.

[0024] 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.

[0025] Preferably, the alkali solution in S2 is at least one of sodium methoxide solution, lithium hydroxide solution and sodium hydroxide solution.

[0026] 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.

[0027] 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.

[0028] 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.

[0029] 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.

[0030] Preferably, the solvent used for extraction in S2 is dichloromethane.

[0031] 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.

[0032] Preferably, the oxidant system in S3 is at least one of 2,2,6,6 - tetramethylpiperidine oxide / potassium bromide / sodium hypochlorite solution, Jones reagent, and Collins reagent.

[0033] More preferably, if the oxidant system is 2,2,6,6 - tetramethylpiperidine oxide / potassium bromide / sodium hypochlorite solution, the molar ratio of tert - butyl 18 - hydroxyoctadecanoate to 2,2,6,6 - tetramethylpiperidine oxide used in the preparation of glycoside intermediate is 1:0.005 - 0.02, the molar ratio of 2,2,6,6 - tetramethylpiperidine oxide to potassium bromide is 1:8 - 12, and the available chlorine content of the sodium hypochlorite solution is 6 - 14%.

[0034] More preferably, if the oxidant system is Jones reagent or Collins reagent, the dosage ratio of tert - butyl 18 - hydroxyoctadecanoate to Jones reagent or Collins reagent used in the preparation of glycoside intermediate is 1 mol:0.8 - 1.2 L.

[0035] More preferably, if the oxidant system is Jones reagent or Collins reagent, isopropanol needs to be added to quench the reaction after the reaction ends.

[0036] Preferably, the solvent used to adjust the pH in S3 is hydrochloric acid solution or sodium hydroxide solution. The hydrochloric acid solution is composed of hydrochloric acid and water, and the dosage ratio of hydrochloric acid to water is 1 mol:0.8 - 1.2 L; the sodium hydroxide solution is composed of sodium hydroxide and water, and the dosage ratio of sodium hydroxide to water is 1 mol:0.8 - 1.2 L.

[0037] Preferably, the column used for purification is a modified silica gel column made of modified silica gel.

[0038] Preferably, the solvent used for extraction in S3 is ethyl acetate.

[0039] Preferably, the solvent used for elution in S3 is methanol - dichloromethane solution, which is composed of methanol and dichloromethane, and the volume ratio of methanol to dichloromethane is 1:18 - 20.

[0040] The present invention discloses a preparation method of modified silica gel, specifically as follows: Soak silica gel in hydrochloric acid solution for 20 - 30 h, then wash with water and dry to obtain activated silica gel. Under nitrogen conditions, add N - (acetylglycyl) - 3 - aminopropyltrimethoxysilane, 3 - mercaptopropylmethyldimethoxysilane, and toluene to the activated silica gel, react at 100 - 130 °C for 5 - 8 h, and after the reaction ends, wash and dry to obtain modified silica gel.

[0041] Preferably, the hydrochloric acid solution is composed of hydrochloric acid and water, and the volume ratio of the usage amounts of hydrochloric acid to water is 1:0.8 - 1.2.

[0042] Preferably, the mass ratio of the usage amounts of activated silica gel to N-(acetylglycyl)-3-aminopropyltrimethoxysilane is 1:0.3 - 0.7.

[0043] Preferably, the mass ratio of the usage amounts of N-(acetylglycyl)-3-aminopropyltrimethoxysilane to 3-mercaptopropylmethyldimethoxysilane is 1:0.8 - 1.2.

[0044] Preferably, the usage ratio of activated silica gel to toluene is 1 g:15 - 25 ml.

[0045] Preferably, the toluene and methanol are used for washing in sequence.

[0046] The present invention discloses a preparation method of a modified silica gel column, specifically as follows: First, an eluent is pre-filled into the column, 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 is allowed to flow out to obtain the modified silica gel column.

[0047] Preferably, the eluent is a methanol-dichloromethane solution, the methanol-dichloromethane solution is composed of methanol and dichloromethane, and the volume ratio of the usage amounts of methanol to dichloromethane is 1:18 - 20.

[0048] More preferably, in the process of preparing the modified silica gel in the present invention, on the basis of using N-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane, N-((4-vinylbenzyl)iminodiacetic acid) can also be used to further prepare the modified silica gel. Introducing N-((4-vinylbenzyl)iminodiacetic acid) can construct a synergistic action system with multiple functional groups, further change the properties of the modified silica gel, improve the performance of the modified silica gel, be beneficial to the adsorption of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, and thus improve the separation and purification ability.

[0049] Preferably, the mass ratio of the usage amounts of the modified silica gel intermediate to N-((4-vinylbenzyl)iminodiacetic acid) is 1:0.3 - 0.8.

[0050] The present invention has the following beneficial effects compared with the prior art: The present invention provides a method for preparing a key intermediate of pevitepa. First, 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide reacts with tert-butyl 18-hydroxyoctadecanoate under the action of a catalyst to form a glycoside intermediate. The glycoside intermediate is deprotected by alkali hydrolysis to remove the acetyl protecting group, and finally, the target product tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, namely the key intermediate of pevitepa, is obtained through an oxidation reaction and purification. The preparation method of the present invention can finally obtain the key intermediate of pevitepa with a high yield and purity, and has simple operation and good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only exemplary, and for those of ordinary skill in the art, other implementation drawings can be obtained based on the provided drawings without creative efforts.

[0052] Figure 1 It is a graph of the yield determination results of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0053] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0054] The concepts involved in the present application will be described below with reference to the drawings first. It should be noted here that the following descriptions of each concept are only for making the content of the present application easier to understand, and do not represent a limitation on the protection scope of the present application; at the same time, without conflict, the embodiments and features in the embodiments of the present application can be combined with each other. The present application will be described in detail below with reference to the drawings and in combination with the embodiments.

[0055] The specific meanings of the abbreviations used in the specification and claims are as follows: Example 1: Preparation of Modified Silica Gel: Soak silica gel in hydrochloric acid solution for 24 h, then wash with water and dry to obtain activated silica gel. Under nitrogen conditions, add N-(acetylglycyl)-3-aminopropyltrimethoxysilane, 3-mercaptopropylmethyldimethoxysilane and toluene to the activated silica gel, react at 120 °C for 6 h, after the reaction, wash successively with toluene and methanol, and finally dry to obtain modified silica gel. The hydrochloric acid solution consists of hydrochloric acid and water, and the volume ratio of the usage amounts of hydrochloric acid and water is 1:1. The mass ratio of the usage amounts of activated silica gel and N-(acetylglycyl)-3-aminopropyltrimethoxysilane is 1:0.5. The mass ratio of the usage amounts of N-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane is 1:1. The usage ratio of activated silica gel and toluene is 1 g:20 ml.

[0056] Preparation of Modified Silica Gel Column: First, pre-fill the column with eluent, then pour the modified silica gel soaked with eluent into the column. After the modified silica gel is deposited, apply nitrogen pressure to drive away the residual bubbles in the column, compact the bed layer, and let the eluent flow out to obtain the modified silica gel column. The eluent is a methanol-dichloromethane solution, and the methanol-dichloromethane solution consists of methanol and dichloromethane. The volume ratio of the usage amounts of methanol and dichloromethane is 1:19.

[0057] Preparation of Glycoside Intermediate: Mix tert-butyl 18-hydroxyoctadecanoate and toluene, then under nitrogen conditions, add silver trifluoromethanesulfonate and 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide, stir and react at 25 °C for 12 h. After the reaction, add water to quench the reaction, separate the organic phase, wash with sodium chloride solution, and obtain the glycoside intermediate after drying and concentration. The usage ratio of tert-butyl 18-hydroxyoctadecanoate and toluene is 1 mol:5 L. The molar ratio of the usage amounts of tert-butyl 18-hydroxyoctadecanoate and silver trifluoromethanesulfonate is 1:2. The molar ratio of the usage amounts of tert-butyl 18-hydroxyoctadecanoate and 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide is 1:1. The sodium chloride solution consists of sodium chloride and water, and the usage ratio of sodium chloride and water is 1 mol:1.12 L.

[0058] 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.

[0059] Example 2: The preparation of modified silica gel is the same as in Example 1.

[0060] The preparation of the modified silica gel column was the same as in Example 1.

[0061] 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.

[0062] 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.

[0063] Example 3: The preparation of the modified silica gel is the same as that in Example 1.

[0064] The preparation of the modified silica gel column is the same as that in Example 1.

[0065] Preparation of the glycoside intermediate: Mix tert-butyl 18-hydroxyoctadecanoate and toluene, then under nitrogen atmosphere, add Sn(OTf)₂ and 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide, and stir at 25 °C for 12 h. After the reaction, quench the reaction with water, separate the organic phase, wash it with sodium chloride solution, and dry and concentrate to obtain the glycoside intermediate. The dosage ratio of tert-butyl 18-hydroxyoctadecanoate to toluene is 1 mol: 5 L, the molar ratio of tert-butyl 18-hydroxyoctadecanoate to Sn(OTf)₂ is 1: 1.5, the molar ratio of tert-butyl 18-hydroxyoctadecanoate to 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide is 1: 1, the sodium chloride solution consists of sodium chloride and water, and the dosage ratio of sodium chloride to water is 1 mol: 1.12 L.

[0066] Preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate: In this example, compared with Example 1 in the preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, the difference is that the glycoside intermediate is the glycoside intermediate prepared in this example, and other conditions and parameters are the same as those in Example 1.

[0067] Example 4: The preparation of the modified silica gel is the same as that in Example 1.

[0068] The preparation of the modified silica gel column is the same as that in Example 1.

[0069] Preparation of the glycoside intermediate: Mix tert-butyl 18-hydroxyoctadecanoate and toluene, then under nitrogen atmosphere, add mercuric chloride and 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide, and stir at 25 °C for 12 h. After the reaction, quench the reaction with water, separate the organic phase, wash it with sodium chloride solution, and dry and concentrate to obtain the glycoside intermediate. The dosage ratio of tert-butyl 18-hydroxyoctadecanoate to toluene is 1 mol: 5 L, the molar ratio of tert-butyl 18-hydroxyoctadecanoate to mercuric chloride is 1: 1, the molar ratio of tert-butyl 18-hydroxyoctadecanoate to 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide is 1: 1, the sodium chloride solution consists of sodium chloride and water, and the dosage ratio of sodium chloride to water is 1 mol: 1.12 L.

[0070] 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.

[0071] Example 5: The preparation of modified silica gel is the same as in Example 1.

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

[0073] The preparation of the glycoside intermediate was the same as in Example 1.

[0074] 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.

[0075] Example 6: The preparation of modified silica gel is the same as in Example 1.

[0076] The preparation of the modified silica gel column was the same as in Example 1.

[0077] The preparation of the glycoside intermediate was the same as in Example 1.

[0078] Preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate: In this example, compared with Example 5 for the preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, the difference lies in that Jones reagent is replaced by Collins reagent, and other conditions and parameters are the same as those in Example 5.

[0079] Example 7: Preparation of modified silica gel: In this example, compared with Example 1 for the preparation of the modified silica gel column, the difference lies in that the mass ratio of the activated silica gel to N-(acetylglycyl)-3-aminopropyltrimethoxysilane used is 1:0.8, and other conditions and parameters are the same as those in Example 1.

[0080] Preparation of the modified silica gel column: In this example, compared with Example 1 for the preparation of the modified silica gel column, the difference lies in that the modified silica gel is replaced by the modified silica gel prepared in this example, and other conditions and parameters are the same as those in Example 1.

[0081] The preparation of the glycoside intermediate is the same as that in Example 1.

[0082] Preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate: In this example, compared with Example 1 for the preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, the difference lies in that the modified silica gel column is the modified silica gel column prepared in this example, and other conditions and parameters are the same as those in Example 1.

[0083] Example 8: Preparation of Modified Silica Gel: Soak silica gel in hydrochloric acid solution for 24 h, then wash with water and dry to obtain activated silica gel. Under nitrogen condition, add N-(acetylglycyl)-3-aminopropyltrimethoxysilane, 3-mercaptopropylmethyldimethoxysilane and toluene to the activated silica gel, and react at 120 °C for 6 h. After the reaction, wash with toluene and methanol in sequence, and then dry to obtain the intermediate of modified silica gel. Add acetonitrile, azobisisobutyronitrile and N-((4-vinylbenzyl)iminodiacetic acid) to the intermediate of modified silica gel, and react at 80 °C for 36 h. After the reaction, wash with acetonitrile and ether in sequence, and finally dry to obtain modified silica gel. The hydrochloric acid solution is composed of hydrochloric acid and water, and the volume ratio of the usage amount of hydrochloric acid to water is 1:1. The mass ratio of the usage amount of activated silica gel to N-(acetylglycyl)-3-aminopropyltrimethoxysilane is 1:1. The mass ratio of the usage amount of N-(acetylglycyl)-3-aminopropyltrimethoxysilane to 3-mercaptopropylmethyldimethoxysilane is 1:1. The usage ratio of activated silica gel to toluene is 1 g:20 ml. The usage ratio of the intermediate of modified silica gel to acetonitrile is 1 g:15 ml. The mass ratio of the usage amount of the intermediate of modified silica gel to azobisisobutyronitrile is 1:0.008. The mass ratio of the usage amount of the intermediate of modified silica gel to N-((4-vinylbenzyl)iminodiacetic acid) is 1:0.38.

[0084] Preparation of Modified Silica Gel Column: In this example, the preparation of the modified silica gel column is different from that in Example 1 in that the modified silica gel is replaced with the modified silica gel prepared in this example, and other conditions and parameters are the same as those in Example 1.

[0085] The preparation of the glycoside intermediate is the same as that in Example 1.

[0086] Preparation of tert-Butyl 18-([β-D-Glucuronosyl-1-yl]oxy)octadecanoate: In this example, the preparation of tert-Butyl 18-([β-D-Glucuronosyl-1-yl]oxy)octadecanoate is different from that in Example 1 in that the modified silica gel column is the modified silica gel column prepared in this example, and other conditions and parameters are the same as those in Example 1.

[0087] Example 9: Preparation of Modified Silica Gel: In this example, the preparation of the modified silica gel column is different from that in Example 8 in that the mass ratio of the usage amount of the intermediate of modified silica gel to N-((4-vinylbenzyl)iminodiacetic acid) is 1:0.6, and other conditions and parameters are the same as those in Example 8.

[0088] Preparation of Modified Silica Gel Column: In this example, the preparation of the modified silica gel column is different from that in Example 8 in that the modified silica gel is replaced with the modified silica gel prepared in this example, and other conditions and parameters are the same as those in Example 8.

[0089] The preparation of the glycoside intermediate was the same as that in Example 8.

[0090] Preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate: In this example, compared with Example 8 in the preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, the difference was that the modified silica gel column was the modified silica gel column prepared in this example, and other conditions and parameters were the same as those in Example 8.

[0091] Comparative Example 1: Preparation of modified silica gel: In this comparative example, compared with Example 1 in the preparation of the modified silica gel column, the difference was that the mass ratio of the activated silica gel to the use amount of N-(acetylglycyl)-3-aminopropyltrimethoxysilane was 1:0.05, and other conditions and parameters were the same as those in Example 1.

[0092] Preparation of modified silica gel column: In this comparative example, compared with Example 1 in the preparation of the modified silica gel column, the difference was that the modified silica gel was replaced with the modified silica gel prepared in this example, and other conditions and parameters were the same as those in Example 1.

[0093] The preparation of the glycoside intermediate was the same as that in Example 1.

[0094] Preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate: In this comparative example, compared with Example 1 in the preparation of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, the difference was that the modified silica gel column was the modified silica gel column prepared in this example, and other conditions and parameters were the same as those in Example 1.

[0095] In the experimental examples of the present invention, the detection of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate was carried out by HPLC method.

[0096] The HPLC method was as follows: Reference substance solution: Accurately weigh 20 mg of the reference substance of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, place it in a 10 ml volumetric flask, dissolve it with the diluent and make the volume up to a solution containing 2 mg per 1 ml as the reference substance solution. The diluent was an acetonitrile aqueous solution, and the acetonitrile aqueous solution was mixed by acetonitrile and water in a volume ratio of 1:1.

[0097] Test solution: Accurately weigh 20 mg of the test sample, place it in a 10 ml volumetric flask, dissolve it with the diluent and make the volume up to a solution containing 2 mg per 1 ml as the test solution. The test sample was tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate prepared in each example or comparative example. The diluent was an acetonitrile aqueous solution, and the acetonitrile aqueous solution was mixed by acetonitrile and water in a volume ratio of 1:1.

[0098] Mobile phase: An aqueous phosphoric acid solution is used as mobile phase A, and acetonitrile is used as mobile phase B. In the preparation of the aqueous phosphoric acid solution, 1 ml of phosphoric acid is accurately measured and placed in a 1000 ml volumetric flask, diluted to volume with water and shaken well, and filtered through a 0.45 μm microporous membrane.

[0099] Detection method: Agilent ZORBAX SB-C18 5 μm 4.6 mm×250 mm is used as the chromatographic column, the detection wavelength is 210 nm, an aqueous phosphoric acid solution is used as mobile phase A, and acetonitrile is used as mobile phase B for gradient elution. The gradient elution program is as follows: at 0 minute, mobile phase A is 40 vol%, and mobile phase B is 60 vol%; at 35 minutes, mobile phase A is 10 vol%, and mobile phase B is 90 vol%; at 50 minutes, mobile phase A is 10 vol%, and mobile phase B is 90 vol%; at 51 minutes, mobile phase A is 40 vol%, and mobile phase B is 60 vol%; at 60 minutes, mobile phase A is 40 vol%, and mobile phase B is 60 vol%; the column temperature is 30 °C, the flow rate is 1.0 ml / min, and the injection volume of the test solution is 20 μL. The resolution between the main peak and each impurity should not be less than 1.5.

[0100] Experimental example 1: Determination of yield: The tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate prepared in Examples 1-10 and Comparative Example 1 was statistically calculated to obtain the yield.

[0101] The results of the yield determination of the tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate prepared in Examples 1-10 and Comparative Example 1 are as Figure 1As shown, compared with Examples 2-4, Example 1 shows that when silver trifluoromethanesulfonate is used as the catalyst, the yield of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate is the highest; compared with Examples 5-6, Example 1 shows that when the oxidant system is 2,2,6,6-tetramethylpiperidine N-oxide, potassium bromide and sodium hypochlorite, the yield of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate is the highest; compared with Example 7, Example 1 shows that the increase in the amount of N-(acetylglycyl)-3-aminopropyltrimethoxysilane within a certain range can improve the effect of the modified silica gel, thereby increasing the yield of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate; compared with Example 8, Example 7 shows that on the basis of using N-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane, the use of N-((4-vinylbenzyl)iminodiacetic acid) to prepare the modified silica gel can further increase the yield of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate; compared with Example 9, Example 8 shows that the increase in the amount of N-((4-vinylbenzyl)iminodiacetic acid) can also increase the yield of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate prepared subsequently; compared with Comparative Example 1, Example 1 shows that the amount of N-(acetylglycyl)-3-aminopropyltrimethoxysilane needs to be within a suitable range, and too low an amount has no obvious effect on increasing the yield of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate.

[0102] Experimental Example 2: Determination of purity: The tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate prepared in Examples 1-10 and Comparative Example 1 was analyzed by HPLC to obtain the purity.

[0103] Table 1 Results of purity determination

[0104] The purity determination results of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate prepared in Examples 1-10 and Comparative Example 1 are shown in Table 1. Comparing Example 1 with Examples 2-4, it shows that when silver trifluoromethanesulfonate is used as a catalyst, the purity of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate is the highest; comparing Example 1 with Examples 5-6, it shows that when the oxidant system is 2,2,6,6-tetramethylpiperidine N-oxide, potassium bromide and sodium hypochlorite, the purity of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate is the highest; comparing Example 1 with Example 7, it shows that within a certain range, the increase in the amount of N-(acetylglycyl)-3-aminopropyltrimethoxysilane used can improve the effect of modified silica gel, thereby increasing the purity of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate; comparing Example 7 with Example 8, it shows that on the basis of using N-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane, the use of N-((4-vinylbenzyl)iminodiacetic acid) to prepare modified silica gel can further increase the purity of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate; comparing Example 8 with Example 9, it shows that the increase in the amount of N-((4-vinylbenzyl)iminodiacetic acid) used can also increase the purity of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate prepared subsequently; comparing Example 1 with Comparative Example 1, it shows that the amount of N-(acetylglycyl)-3-aminopropyltrimethoxysilane used needs to be within a suitable range, and too low an amount has no obvious effect on improving the purity of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate.

[0105] Experimental Example 3: Determination of the adsorption rate of modified silica gel for tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate. Add a solution of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate to activated silica gel, shake at 30 °C and 150 rpm for uniform adsorption, and determine the content of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate in the solution by HPLC to determine the time required for adsorption to reach equilibrium. Adsorption rate = (concentration of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate in the solution before adsorption - concentration of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate in the solution after adsorption) / concentration of tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy) octadecanoate in the solution before adsorption × 100%.

[0106] Table 2 Determination results of adsorption rate

[0107] The results are shown in Table 2. Comparing Example 1 with Example 7 indicates that an increase in the amount of nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane within a certain range can improve the adsorption of the modified silica gel. Comparing Example 7 with Example 8 shows that on the basis of using nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane, the use of N-((4-vinylbenzyl)iminodiacetic acid) to prepare the modified silica gel can further improve the adsorption of the prepared modified silica gel. Comparing Example 8 with Example 9 shows that an increase in the amount of N-((4-vinylbenzyl)iminodiacetic acid) can also improve the adsorption of the modified silica gel. Comparing Example 1 with Comparative Example 1 shows that the amount of nitrogen-(acetylglycyl)-3-aminopropyltrimethoxysilane needs to be within a suitable range, and too low an amount has no obvious effect on improving the adsorption rate of the modified silica gel.

[0108] The above-described embodiments and / or implementation manners are only used to illustrate the preferred embodiments and / or implementation manners for implementing the technology of the present invention, and do not impose any formal restrictions on the implementation manners of the technology of the present invention. Any person skilled in the art, without departing from the scope of the technical means disclosed in the content of the present invention, may make some changes or modifications to other equivalent embodiments, but should still be regarded as the same technology or embodiment as the present invention in essence.

[0109] Specific examples are used in this article to elaborate on the principles and implementation manners of the present application. The description of the above embodiments is only used to help understand the method and its core idea of the present application. The above is only the preferred implementation manner of the present application. It should be noted that due to the limited nature of written expression and the objectively infinite specific structures, for those of ordinary skill in the art, without departing from the principle of the present application, several improvements, refinements or changes can be made, or the above technical features can be combined in an appropriate manner; these improvements, refinements, changes or combinations, or directly applying the concept and technical solution of the invention to other occasions without improvement, should all be regarded as the protection scope of the present application.

Claims

1. A preparation method of a key intermediate of pervitin, comprising: S1: Mixing tert-butyl 18-hydroxyoctadecanoate with a solvent, and reacting with 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide under nitrogen conditions in the presence of a catalyst to form a glycoside intermediate; the molar ratio of the amount of tert-butyl 18-hydroxyoctadecanoate to 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide used is 1:0.8 - 1.2; S2: Mixing the glycoside intermediate prepared in S1 with a solvent, and then subjecting it to an alkali hydrolysis reaction to obtain an oil; S3: Adding a solvent to the oil prepared in S2, and oxidizing it under the action of an oxidant system to form a crude product, and the crude product is purified by column chromatography to obtain tert-butyl 18-([β-D-glucuronosyl-1-yl]oxy)octadecanoate, which is the key intermediate of pervitin.

2. The preparation method of a key intermediate of pevicetide according to claim 1, characterized in that: The solvent in S1 is toluene, and the dosage ratio of tert-butyl 18-hydroxyoctadecanoate to the solvent is 1 mol: 4 - 6 L.

3. The preparation method of a key intermediate of pevicetide according to claim 1, characterized in that: The catalyst in S1 is a metal salt, a Lewis acid or a phase transfer agent, and the molar ratio of the amount of tert-butyl 18-hydroxyoctadecanoate to the catalyst used is 1:1 - 6.

4. The preparation method of a key intermediate of pevicetide according to claim 1, characterized in that: The solvent in S2 is methanol, and the dosage ratio of tert-butyl 18-hydroxyoctadecanoate when preparing the glycoside intermediate to the solvent when preparing the oil is 1 mol: 4 - 6 L.

5. The preparation method of a key intermediate of peviteptide according to claim 1, characterized in that: The alkali solution in S2 is at least one of a sodium methoxide solution, a lithium hydroxide solution and a sodium hydroxide solution, and the pH is adjusted to 9 - 11 with the alkali.

6. The preparation method of a key intermediate of pevicetide according to claim 1, characterized in that: The solvent in S3 is at least one of 1,4-dioxane, tetrahydrofuran and methyl tert-butyl ether, and the dosage ratio of tert-butyl 18-hydroxyoctadecanoate when preparing the glycoside intermediate to the solvent when preparing the crude product is 1 mol: 4 - 6 L.

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

8. The preparation method of a key intermediate of pevibatinib according to claim 3, characterized in that: 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, N-(acetylglycyl)-3-aminopropyltrimethoxysilane and 3-mercaptopropylmethyldimethoxysilane.

9. The preparation method of a key intermediate of pevicetide according to claim 1, characterized in that: The reaction temperature in S1 is 20 - 30 °C, and the reaction time is 10 - 15 h; the reaction temperature in S2 is 20 - 30 °C, and the reaction time is 3 - 5 h; the reaction temperature in S3 is 5 - 15 °C, and the reaction time is 3 - 5 h.

10. Use of the key intermediate of pervitin prepared by any of the methods of claims 1 - 9 in the preparation of pervitin.

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