Polyhydroxypyrrolidine ether compounds, methods of making and using the same

Abstract

This invention relates to the field of organic synthesis technology, specifically to a polyhydroxypyrrolidone ether compound, its preparation method, and its application. The structure of the polyhydroxypyrrolidone ether compound is shown in formula (1), and each group in formula (1) has a specific selection; the stereoconfiguration of the carbon atoms at positions 2, 3, and 4 in formula (1) is "2R,3R,4R" or "2S,3S,4S". The polyhydroxypyrrolidone ether compound of this invention and its pharmaceutically acceptable salts exhibit excellent selectivity and strong inhibitory effects on β-glucosidase, and are suitable for the prevention or treatment of Gaucher's disease and Parkinson's syndrome. Formula (1), Formula (2).

Description

Technical Field

[0001] This invention relates to the field of organic synthesis technology, specifically to a polyhydroxypyrrolidone ether compound, its preparation method, and its application. Background Technology

[0002] Lysosomal acid β-glucosidase (GCase) is a lysosomal enzyme encoded by the GBA1 gene that hydrolyzes glycosphingolipid substrates such as glucosceramide (GluCer) and glucosphoside (GluSph). Impaired catalytic function and stability of GCase are the cause of Gaucher disease (GD), the most common lysosomal storage disorder (LSD).

[0003] Molecular chaperones can act as templates for folding, specifically binding to the catalytic domain of GCases, inducing the correct conformation of misfolded enzymes, and ensuring their subsequent normal transport and function. Iminosaccharide-type GCase inhibitors are among the most representative and rapidly developing molecular chaperones [Sánchez-Fernández, EM; García Fernández, JM; Mellet, CO. Glycomimetic-based pharmacological chaperones for lysosomalstorage disorders: lessons from Gaucher, GM1-gangliosidosis and Fabrydiseases]. Chem. Commun. 2016, 52 , 5497-5515.].

[0004] Furthermore, recent studies have shown that insufficient GCase activity poses a significant risk of Parkinson's disease, especially in the elderly, where the risk is markedly increased. This gives GCase inhibitor-based molecular chaperones broader research value and application potential. Therefore, iminosaccharide-type polyhydroxypyrrolidone ethers have important application value in the treatment of lysosomal storage diseases, Gaucher's disease, and Parkinson's disease. Summary of the Invention

[0005] The purpose of this invention is to overcome the problems existing in the prior art and provide a polyhydroxypyrrolidone compound, its preparation method, and its application. This polyhydroxypyrrolidone compound and its pharmaceutically acceptable salt exhibit excellent selectivity and strong inhibitory effects on β-glucosidase, making it suitable for the prevention or treatment of Gaucher's disease and Parkinson's syndrome.

[0006] To achieve the above objectives, the present invention provides a first aspect of a polyhydroxypyrrolidone ether compound and a pharmaceutically acceptable salt thereof, the structure of which is shown in formula (1): Equation (1) ; Among them, R 1 One or more alkyl groups selected from C5-C30; R 2 and R 3 Simultaneously selected from H and one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more alkyl groups and halogens of C1-C12; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (1) is “2R,3R,4R” or “2S,3S,4S”.

[0007] A second aspect of the present invention provides a Boc-protected pyrrolidone compound and a pharmaceutically acceptable salt thereof, the structure of which is shown in formula (2): Equation (2) ; Among them, R 1 One or more alkyl groups selected from C5-C30, R2 and R3 are both selected from one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more alkyl groups and halogens from C1-C12; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is “2R,3R,4R” or “2S,3S,4S”.

[0008] A third aspect of this invention provides a method for preparing polyhydroxypyrrolidone ether compounds and pharmaceutically acceptable salts thereof, the method comprising: Scenario 1: When R 2 and R 3 When H is present, in the presence of a first catalyst and a first acidic reagent, hydrogen is used to reduce the Boc protected pyrrolidone ether compound shown in formula (2) to obtain the polyhydroxypyrrolidone ether compound shown in formula (1). Scenario 2: When R 2 and R 3 When one or more of the substituted or unsubstituted benzyl groups are present, the Boc-protected pyrrolidone ethers shown in formula (2) are subjected to a deBoc reaction in the presence of a second acidic reagent to obtain the polyhydroxypyrrolidone ethers shown in formula (1). Equation (2) Equation (1) ; Among them, R 1 R 2 R 3R2 and R3 are as defined in the first and second aspects above; the stereo configurations of the carbon atoms at positions 2, 3 and 4 in equation (2) are “2R,3R,4R” or “2S,3S,4S”; the stereo configurations of the carbon atoms at positions 2, 3 and 4 in equation (1) are “2R,3R,4R” or “2S,3S,4S”.

[0009] A fourth aspect of the present invention provides a method for preparing Boc-protected pyrrolidine ether compounds and pharmaceutically acceptable salts thereof, the method comprising: reacting a hydroxyl-containing Boc-protected pyrrolidine compound of formula (3) with R in the presence of a second catalyst and a basic reagent. 1 X undergoes an etherification reaction to obtain the Boc protected pyrrolidine ether compound shown in formula (2); Equation (3) Equation (2) ; Among them, R 1 R2 and R3 are as defined in the second aspect above, and X is selected from one or more halogens; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (3) is “2R,3R,4R” or “2S,3S,4S”; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is “2R,3R,4R” or “2S,3S,4S”.

[0010] The fifth aspect of the present invention provides a glycosidase inhibitor comprising the above-described polyhydroxypyrrolidone ether compounds and their pharmaceutically acceptable salts.

[0011] The sixth aspect of the present invention provides a small molecule drug companion comprising the above-mentioned polyhydroxypyrrolidone ether compounds and their pharmaceutically acceptable salts.

[0012] The seventh aspect of the present invention provides the use of the above-mentioned polyhydroxypyrrolidone ether compounds and pharmaceutically acceptable salts thereof in the prevention or treatment of Gaucher disease or Parkinson's syndrome.

[0013] This invention utilizes a special structural design and specific preparation method to obtain a polyhydroxypyrrolidone ether compound and its intermediates that exhibit excellent selectivity and strong inhibitory effect against β-glucosidase. This polyhydroxypyrrolidone ether compound, in particular, demonstrates excellent selectivity and inhibitory ability against β-glucosidase derived from human lysosomes. It also exhibits good molecular chaperone activity against β-glucosidase derived from lysosomes in human N370S fibroblasts, thus making it suitable for the prevention or treatment of Gaucher's disease and Parkinson's syndrome, achieving good preventive or therapeutic effects. Attached Figure Description

[0014] Figure 1 The graph shows the change in enzyme deformation with temperature obtained from test example 2; Figure 2The graph shows the degree to which the small molecule chaperone obtained in Test Example 3 enhances the activity of residual enzymes in human N370S fibroblasts. Detailed Implementation

[0015] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0016] In this invention, the alkyl groups used in expressions such as "C5-C30 alkyl" and "C1-C12 alkyl" can be either straight-chain alkyl or branched-chain alkyl. The same applies to other similar expressions.

[0017] In this invention, "substituted or unsubstituted benzyl" means that the H on the benzyl group can be substituted by a substituent or not; when substituted, the substituted H can be one or more, and when multiple H are substituted, they can be H on the same carbon atom or H on different carbon atoms. Other similar or identical expressions are analogous.

[0018] In this invention, The numbers "1, 2, 3, 4, 5" in the structure indicate the position of the carbon atom, while the numbers in "2R, 3R, 4R" and "2S, 3S, 4S" that indicate the stereoconfiguration of the compound refer to the carbon atom at the corresponding position. R or S refers to the absolute configuration of the carbon atom at the corresponding position. For example, "2R" indicates that the carbon atom at position 2 in the compound has an absolute configuration of R, and "4S" indicates that the carbon atom at position 4 in the compound has an absolute configuration of S.

[0019] In this invention, some groups in the compound structure can be represented by their abbreviations, such as "Boc" for tert-butyloxycarbonyl and "Bn" for benzyl.

[0020] The present invention provides a first aspect of a polyhydroxypyrrolidone ether compound and a pharmaceutically acceptable salt thereof, the structure of which is shown in formula (1): Equation (1) ; Among them, R 1 One or more alkyl groups selected from C5-C30; R 2 and R 3Simultaneously selected from H and one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more alkyl groups and halogens of C1-C12; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (1) is “2R,3R,4R” or “2S,3S,4S”.

[0021] According to the present invention, by designing the structure of polyhydroxypyrrolidone ether compounds, they can be made to have selective and inhibitory effects on β-glucosidase. To obtain higher activity, the structure of the polyhydroxypyrrolidone ether compounds can be further selected. Preferably, R... 1 One or more alkyl groups selected from C5-C24; R 2 and R 3 Simultaneously selected from H and one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more alkyl groups and halogens of C1-C6; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (1) is “2R,3R,4R” or “2S,3S,4S”.

[0022] More preferably, R 1 Selected from one or more n-alkyl groups from C5-C18; R 2 and R 3 Simultaneously selected from H and one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more of C1-C3 alkyl groups, F, Cl and Br; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (1) is “2R,3R,4R” or “2S,3S,4S”.

[0023] More preferably, R 1 Selected from one or more of n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecanyl, n-hexadecyl, n-heptadecyl, and n-octadecyl; R 2 and R 3 Simultaneously selected from one or more of H, benzyl, p-methylbenzyl, p-ethylbenzyl, p-isopropylbenzyl, p-chlorobenzyl and p-bromobenzyl; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (1) is "2R,3R,4R" or "2S,3S,4S".

[0024] According to a preferred embodiment of the present invention, the polyhydroxypyrrolidone ether compound is selected from one or more compounds shown in the following formula:

[0025]

[0026]

[0027]

[0028]

[0029] .

[0030] According to a preferred embodiment of the present invention, the pharmaceutically acceptable salts of the polyhydroxypyrrolidone ether compounds include one or more of hydrochloride, sulfonate, trifluoroacetate, carbonate, nitrate, phosphate, folic acid, and acetate, preferably one or more of hydrochloride, sulfonate, and trifluoroacetate.

[0031] A second aspect of the present invention provides a Boc-protected pyrrolidone compound and a pharmaceutically acceptable salt thereof, the structure of which is shown in formula (2): Equation (2) ; Among them, R 1 One or more alkyl groups selected from C5-C30, R2 and R3 are both selected from one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more alkyl groups and halogens from C1-C12; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is “2R,3R,4R” or “2S,3S,4S”.

[0032] According to the present invention, Boc-protected pyrrolidone compounds, as intermediates in the preparation of polyhydroxypyrrolidone compounds, can be further selected in structure to obtain polyhydroxypyrrolidone compounds with higher selectivity and better inhibitory effect against β-glucosidase. Preferably, R 1 One or more alkyl groups selected from C5-C24; R2 and R3 are both selected from one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more alkyl groups and halogens from C1-C6; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is “2R,3R,4R” or “2S,3S,4S”.

[0033] More preferably, R 1 One or more of the C5-C18 n-alkyl groups; R2 and R3 are both selected from one or more of the substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more of the C1-C3 alkyl groups, F, Cl and Br; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is “2R,3R,4R” or “2S,3S,4S”.

[0034] More preferably, R 1 R2 is selected from one or more of n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecanyl, n-hexadecyl, n-heptadecyl, and n-octadecyl; R2 and R3 are simultaneously selected from one or more of benzyl, p-methylbenzyl, p-ethylbenzyl, p-isopropylbenzyl, p-chlorobenzyl, and p-bromobenzyl; the stereoconfiguration of the carbon atoms at positions 2, 3, and 4 in formula (2) is "2R,3R,4R" or "2S,3S,4S".

[0035] According to a preferred embodiment of the present invention, the Boc-protected pyrrolidone compound is selected from one or more compounds shown in the following formula:

[0036]

[0037]

[0038]

[0039]

[0040] .

[0041] According to a preferred embodiment of the present invention, the pharmaceutically acceptable salt of the Boc protected pyrrolidone compound includes one or more of hydrochloride, sulfonate, trifluoroacetate, carbonate, nitrate, phosphate, folic acid, and acetate, preferably one or more of hydrochloride, sulfonate, and trifluoroacetate.

[0042] A third aspect of this invention provides a method for preparing polyhydroxypyrrolidone ether compounds and pharmaceutically acceptable salts thereof, the method comprising: Scenario 1: When R 2 and R 3 When H is present, in the presence of a first catalyst and a first acidic reagent, hydrogen is used to reduce the Boc protected pyrrolidone ether compound shown in formula (2) to obtain the polyhydroxypyrrolidone ether compound shown in formula (1). Scenario 2: When R 2 and R 3 When one or more of the substituted or unsubstituted benzyl groups are present, the Boc-protected pyrrolidone ethers shown in formula (2) are subjected to a deBoc reaction in the presence of a second acidic reagent to obtain the polyhydroxypyrrolidone ethers shown in formula (1). Equation (2) Equation (1) ; Among them, R 1 R 2 R 3 R2 and R3 are as defined in the first and second aspects above; the stereo configurations of the carbon atoms at positions 2, 3 and 4 in equation (2) are “2R,3R,4R” or “2S,3S,4S”; the stereo configurations of the carbon atoms at positions 2, 3 and 4 in equation (1) are “2R,3R,4R” or “2S,3S,4S”.

[0043] According to the present invention, in order to accelerate the reaction rate and obtain a better reduction effect, the first catalyst is preferably selected from one or more of Raney nickel, Pd / C, palladium black, palladium hydroxide, palladium acetate, palladium chloride, platinum chloride and platinum black, and more preferably from one or more of Raney nickel, Pd / C, palladium hydroxide and palladium acetate.

[0044] Preferably, relative to 1g of the Boc protected pyrrolidone compound, the amount of the first catalyst is 0.005-0.5g, more preferably 0.01-0.1g, for example, it can be 0.01g, 0.03g, 0.05g, 0.08g and 0.1g, or any value between these values.

[0045] According to the present invention, in order to facilitate the reaction and better remove Boc, the first acidic reagent is preferably selected from one or more of hydrochloric acid, sulfuric acid, p-toluenesulfonic acid and trifluoroacetic acid, and more preferably hydrochloric acid and / or sulfuric acid.

[0046] Preferably, the first acidic reagent is provided in the form of a first acid solution. The solvent for the first acid solution may be selected from suitable solvents such as methanol, ethanol, and tetrahydrofuran.

[0047] Preferably, the concentration of the first acid solution is 1-10N, more preferably 2-8N, for example, it can be 2N, 5N, 6N, and 8N, or any range between these values. Wherein, N is the equivalent concentration, that is, the number of gram equivalents of solute contained in 1L of solution. For example, when the first acid solution is hydrochloric acid, 1N = 1 mol / L; when the first acid solution is sulfuric acid, 1N = 0.5 mol / L.

[0048] Preferably, relative to 1g of the Boc protected pyrrolidone compound, the amount of the first acid solution is 0.5-5mL, preferably 0.8-2mL, for example, it can be 0.8mL, 1.2mL, 1.5mL and 2mL and any range between these values.

[0049] According to the present invention, in order to provide a suitable reaction environment, the solvent for the reduction treatment is preferably selected from one or more of methanol, ethanol, acetonitrile, dichloromethane, chloroform, tetrahydrofuran, diethyl ether, ethyl acetate, acetic acid, water, 1,4-dioxane and N,N-dimethylformamide, and more preferably one or more of methanol, ethanol and acetonitrile.

[0050] Preferably, relative to 1g of the Boc protected pyrrolidone compound, the amount of solvent used for the reduction treatment is 10-120mL, preferably 40-80mL, for example, it can be 40mL, 50mL, 60mL and 80mL and any range between these values.

[0051] According to the present invention, in order to achieve better reaction results and improve reaction efficiency, preferably, the conditions for the reduction treatment include: hydrogen pressure of 1-10 atm, temperature of 10-60°C, and time of 2-16 h. More preferably, the conditions for the reduction treatment include: hydrogen pressure of 1-5 atm (e.g., values ​​such as 1 atm, 2 atm, 3 atm, and 5 atm, and any range thereof), temperature of 20-50°C (e.g., values ​​such as 20°C, 30°C, 40°C, and 50°C, and any range thereof), and time of 5-10 h (e.g., values ​​such as 5 h, 8 h, 9 h, and 10 h, and any range thereof).

[0052] According to the present invention, in order to obtain a high-purity product, after the reduction treatment is completed, the obtained reaction solution may be subjected to a first post-treatment. Preferably, the first post-treatment includes: purging with an inert gas (e.g., nitrogen or argon), filtering the reaction solution, removing the solvent by rotary evaporation, and obtaining the polyhydroxypyrrolidone ether compound.

[0053] According to the present invention, in order to better remove Boc and improve reaction efficiency, the second acidic reagent is preferably selected from one or more of hydrochloric acid, sulfuric acid, p-toluenesulfonic acid and trifluoroacetic acid, and more preferably hydrochloric acid and / or sulfuric acid.

[0054] Preferably, the second acidic reagent is provided in the form of a second acid solution. The solvent for the second acid solution may be selected from suitable solvents such as methanol, ethanol, and tetrahydrofuran.

[0055] Preferably, the concentration of the second acid solution is 1-10N, more preferably 2-8N, for example, it can be 2N, 5N, 6N and 8N and any range between these values.

[0056] Preferably, relative to 1g of the Boc protected pyrrolidone compound, the amount of the second acid solution is 0.5-5mL, preferably 0.8-2mL, for example, it can be 0.8mL, 1.2mL, 1.5mL and 2mL and any range between these values.

[0057] According to the present invention, in order to ensure sufficient contact between the reaction substrate and provide a favorable reaction environment, the solvent for the deBoc reaction is preferably selected from one or more of methanol, ethanol, acetonitrile, dichloromethane, chloroform, tetrahydrofuran, diethyl ether, ethyl acetate, acetic acid, water, 1,4-dioxane and N,N-dimethylformamide, and more preferably one or more of methanol, ethanol and acetonitrile.

[0058] Preferably, relative to 1g of the Boc-protected pyrrolidone compound, the amount of solvent used in the Boc removal reaction is 10-120mL, preferably 40-80mL, for example, it can be 40mL, 50mL, 60mL and 80mL and any range between these values.

[0059] According to the present invention, in order to improve the reaction yield and obtain better Boc removal reaction results, preferably, the conditions for the Boc removal reaction include: a temperature of 10-60°C and a time of 2-16 h. More preferably, the conditions for the Boc removal reaction include: a temperature of 20-50°C (e.g., values ​​such as 20°C, 30°C, 40°C, and 50°C, or any range thereof), and a time of 5-10 h (e.g., values ​​such as 5 h, 8 h, 9 h, and 10 h, or any range thereof). Preferably, the Boc removal reaction is carried out in a non-reactive gas atmosphere, such as nitrogen or argon.

[0060] According to the present invention, in order to obtain a high-purity product, after the Boc removal reaction is completed, the obtained reaction solution can be subjected to a second post-treatment. Preferably, the second post-treatment includes: removing the solvent from the reaction solution to obtain the polyhydroxypyrrolidone ether compound.

[0061] A fourth aspect of the present invention provides a method for preparing Boc-protected pyrrolidine ether compounds and pharmaceutically acceptable salts thereof, the method comprising: reacting a hydroxyl-containing Boc-protected pyrrolidine compound of formula (3) with R in the presence of a second catalyst and a basic reagent. 1 X undergoes an etherification reaction to obtain the Boc protected pyrrolidine ether compound shown in formula (2); Equation (3) Equation (2) ; Among them, R 1R2 and R3 are as defined in the second aspect above, and X is selected from one or more halogens; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (3) is “2R,3R,4R” or “2S,3S,4S”; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is “2R,3R,4R” or “2S,3S,4S”.

[0062] According to the present invention, in order to better prepare Boc-protected pyrrolidine ether compounds, preferably, the hydroxyl-containing Boc-protected pyrrolidine compound is selected from one or more compounds shown in the following formula: .

[0063] According to the present invention, in order to improve the efficiency of the etherification reaction and obtain better reaction results, the second catalyst is preferably selected from one or more of tetrabutylammonium iodide, tetrabutylammonium chloride, tetrabutylammonium bromide, tetrabutylammonium fluoride and crown ether, preferably tetrabutylammonium iodide and / or tetrabutylammonium bromide.

[0064] Preferably, relative to 1g of the hydroxyl-containing Boc-protected pyrrolidine compound, the amount of the second catalyst is 0.01-1g, preferably 0.05-0.5g, for example, it can be 0.05g, 0.1g, 0.2g, 0.3g and 0.5g, or any value between these values.

[0065] According to the present invention, in order to provide an alkaline environment and facilitate the reaction, the alkaline reagent is preferably selected from one or more of potassium carbonate, sodium hydride, potassium hydroxide, sodium hydroxide and calcium hydroxide, and more preferably potassium carbonate and / or sodium hydride.

[0066] Preferably, relative to 1g of the hydroxyl-containing Boc-protected pyrrolidine compound, the amount of the alkaline reagent is 0.1-5g, preferably 0.5-2g, for example, it can be 0.5g, 1g, 1.5g and 2g, or any value between these values.

[0067] According to the present invention, in order to obtain the desired product and improve the yield, preferably, relative to 1g of the hydroxyl-containing Boc-protected pyrrolidine compound, R 1 The amount of X is 0.1-10g, preferably 0.5-5g, for example, it can be 0.5g, 1g, 2g, 3g and 5g and any range between these values.

[0068] According to the present invention, in order to make the materials more fully mixed and contacted and achieve better reaction results, the solvent for the etherification reaction is preferably selected from one or more of diethyl ether, tetrahydrofuran, N,N-dimethylformamide, 1,4-dioxane and dichloromethane, preferably tetrahydrofuran and / or N,N-dimethylformamide.

[0069] Preferably, relative to 1g of the hydroxyl-containing Boc-protected pyrrolidine compound, the amount of solvent used in the etherification reaction is 1-1000mL, preferably 200-600mL, for example, it can be 200mL, 350mL, 465mL, 500mL and 600mL and any range between these values.

[0070] According to the present invention, in order to ensure the reaction proceeds fully and to obtain a higher yield, the etherification reaction conditions preferably include: a temperature of 10-50°C and a time of 2-16 h. More preferably, the etherification reaction conditions include: a temperature of 20-40°C (e.g., values ​​such as 20°C, 25°C, 30°C, and 40°C, or any range thereof), and a time of 6-12 h (e.g., values ​​such as 6 h, 8 h, 10 h, and 12 h, or any range thereof). The etherification reaction is preferably carried out in a non-reactive gas atmosphere, such as nitrogen or argon.

[0071] According to the present invention, the specific process of the etherification reaction may, for example, be: adding the hydroxyl-containing Boc-protected pyrrolidine compound and the basic reagent to a solvent, stirring for 20-40 min, and then adding R. 1 X undergoes the etherification reaction to obtain the Boc protected pyrrolidine ether compound.

[0072] According to the present invention, in order to obtain a product with better properties and purity, after the etherification reaction is completed, the obtained reaction solution can be subjected to a third post-treatment. Preferably, the third post-treatment includes: quenching the reaction solution (e.g., quenching with a suitable reagent such as a saturated ammonium chloride aqueous solution), diluting (e.g., diluting with an appropriate amount of water), extracting (e.g., extracting with a suitable solvent such as ethyl acetate), drying the obtained organic phase (e.g., drying with a drying agent such as anhydrous sodium sulfate or anhydrous magnesium sulfate), removing the solvent by rotation, and separating by silica gel column chromatography to obtain the Boc protected pyrrolidone ether compound.

[0073] According to the present invention, the hydroxyl-containing Boc-protected pyrrolidine compound and its pharmaceutically acceptable salt can be prepared commercially available or by referring to methods provided in existing published patents or literature, for example, by the following methods:

[0074] ; Step 1: Combine the compound shown in formula (a) with... The first contact reaction is carried out to obtain the compound shown in formula (b), wherein X1 is selected from Cl or Br, the stereoconfiguration of the carbon atoms at positions 2 and 3 in formula (a) is "2R,3R" or "2S,3S", and the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (b) is "2R,3R,4R" or "2S,3S,4S".

[0075] According to the present invention, in order to obtain the desired product, preferably, relative to 1g of the compound shown in formula (a), The dosage is 2-20 mL, preferably 4-16 mL, for example, it can be 4 mL, 7 mL, 10 mL, 12 mL and 16 mL and any range between these values.

[0076] According to the present invention, in order to facilitate the reaction, the solvent for the first contact reaction is preferably selected from one or more of tetrahydrofuran, diethyl ether, toluene, methyl tert-butyl ether and benzene, preferably tetrahydrofuran and / or diethyl ether.

[0077] Preferably, the amount of solvent used in the first contact reaction is 2-20 mL, more preferably 4-16 mL, relative to 1 g of the compound shown in formula (a), for example, it can be 4 mL, 8 mL, 10 mL, 12 mL and 16 mL and any range between these values.

[0078] According to the present invention, in order to obtain a better reaction rate, preferably, the conditions for the first contact reaction include: a temperature of 10-50°C and a time of 10-60 min. More preferably, the conditions for the first contact reaction include: a temperature of 20-40°C (e.g., values ​​such as 20°C, 25°C, 30°C, and 40°C, or any range thereof), and a time of 20-40 min (e.g., values ​​such as 20 min, 25 min, 30 min, and 40 min, or any range thereof). Preferably, the first contact reaction is carried out in a non-reactive gas atmosphere, such as nitrogen or argon.

[0079] According to the present invention, the specific process of the first contact reaction may be, for example, adding the compound shown in formula (a) to a solvent, and adding it at -10°C to 10°C. The first contact reaction is carried out to obtain the compound shown in formula (b).

[0080] According to the present invention, after the first contact reaction is completed, the obtained reaction solution can be subjected to a fourth post-treatment. Preferably, the fourth post-treatment includes: quenching the reaction solution (e.g., quenching with a suitable reagent such as a saturated ammonium chloride aqueous solution), extraction (e.g., extraction with a suitable solvent such as ethyl acetate), drying the obtained organic phase (e.g., drying with a suitable desiccant such as anhydrous sodium sulfate or anhydrous magnesium sulfate), and then removing the solvent by rotation to obtain the compound shown in formula (b).

[0081] Step 2: In the presence of copper acetate and zinc powder, the compound shown in formula (b) undergoes a second contact reaction to obtain the compound shown in formula (c), wherein the stereoconfiguration of the carbon atoms at positions 2, 3, and 4 in formula (b) is "2R,3R,4R" or "2S,3S,4S", and the stereoconfiguration of the carbon atoms at positions 2, 3, and 4 in formula (c) is "2R,3R,4R" or "2S,3S,4S".

[0082] According to the present invention, in order to obtain a better dehydroxylation effect, preferably, the amount of copper acetate used is 0.005-0.1g, more preferably 0.01-0.05g, relative to 1g of the compound shown in formula (b), for example, it can be 0.01g, 0.025g, 0.034g and 0.05g and any range between these values.

[0083] According to the present invention, in order to obtain better reaction results, preferably, the amount of zinc powder used is 1-5g, more preferably 1.5-3g, relative to 1g of the compound shown in formula (b), for example, it can be 1.5g, 2.2g, 2.8g and 3g and any range between these values.

[0084] According to the present invention, in order for the reaction to proceed smoothly, the solvent for the second contact reaction is preferably glacial acetic acid.

[0085] Preferably, the amount of solvent used in the second contact reaction is 2-20 mL, more preferably 5-15 mL, relative to 1 g of the compound shown in formula (b), for example, it can be 5 mL, 8 mL, 10 mL, 12 mL and 15 mL and any range between these values.

[0086] According to the present invention, in order to obtain a higher yield, preferably, the conditions for the second contact reaction include: a temperature of 10-50°C and a time of 1-8 hours. More preferably, the conditions for the second contact reaction include: a temperature of 20-40°C (e.g., values ​​such as 20°C, 25°C, 30°C, and 40°C, or any range thereof), and a time of 2-6 hours (e.g., values ​​such as 2 hours, 4 hours, 5 hours, and 6 hours, or any range thereof). Preferably, the second contact reaction is carried out in a non-reactive gas atmosphere, such as nitrogen or argon.

[0087] According to the present invention, the specific process of the second contact reaction can be, for example, adding copper acetate and zinc powder to a solvent (glacial acetic acid), adding a glacial acetic acid solution of the compound shown in formula (b) (the concentration can be selected in a wide range, for example, 0.01-1 g / mL), and carrying out the second contact reaction to obtain the compound shown in formula (c).

[0088] According to the present invention, after the second contact reaction is completed, the obtained reaction solution can be subjected to a fifth post-treatment. Preferably, the fifth post-treatment includes: filtering the reaction solution, removing the solvent from the filtrate, and obtaining the compound shown in formula (c).

[0089] Step 3: In the presence of an inorganic base, the compound shown in formula (c) is subjected to a third contact reaction with di-tert-butyl dicarbonate to obtain the compound shown in formula (d), wherein the stereoconfiguration of the carbon atoms at positions 2, 3, and 4 in formula (c) is "2R,3R,4R" or "2S,3S,4S", and the stereoconfiguration of the carbon atoms at positions 2, 3, and 4 in formula (d) is "2R,3R,4R" or "2S,3S,4S".

[0090] According to the present invention, in order to better achieve Boc, the inorganic base is preferably selected from one or more of sodium bicarbonate, potassium bicarbonate, sodium carbonate and potassium carbonate, preferably sodium bicarbonate and / or potassium bicarbonate.

[0091] Preferably, the amount of the inorganic base used relative to 1g of the compound shown in formula (c) is 1-5g, more preferably 1-3g, for example, it can be 1g, 2g, 2.5g and 3g and any range between these values.

[0092] According to the present invention, in order to improve the yield of the reaction, preferably, the amount of di-tert-butyl dicarbonate is 0.5-5g, more preferably 1-3g, relative to 1g of the compound shown in formula (c), for example, it can be 1g, 1.5g, 2g and 3g and any range between these values.

[0093] According to the present invention, in order to provide a favorable reaction environment, the solvent for the third contact reaction is preferably selected from one or more of tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide, dimethyl sulfoxide and toluene, preferably tetrahydrofuran and / or 1,4-dioxane.

[0094] Preferably, the amount of solvent used in the third contact reaction is 2-30 mL, more preferably 5-20 mL, relative to 1 g of the compound shown in formula (c), for example, it can be 5 mL, 8 mL, 15 mL and 20 mL and any range between these values.

[0095] According to the present invention, for better reaction, the conditions for the third contact reaction preferably include: a temperature of 10-50°C and a time of 20-120 min. More preferably, the conditions for the third contact reaction include: a temperature of 20-40°C (e.g., values ​​such as 20°C, 25°C, 30°C, and 40°C, or any range thereof), and a time of 40-80 min (e.g., values ​​such as 40 min, 60 min, 70 min, and 80 min, or any range thereof). Preferably, the third contact reaction is carried out in a non-reactive gas atmosphere, such as nitrogen or argon.

[0096] According to the present invention, the specific process of the third contact reaction can be, for example, adding the compound shown in formula (c) and an inorganic base to a solvent, adding di-tert-butyl dicarbonate at -10°C to 10°C, and carrying out the third contact reaction to obtain the compound shown in formula (d).

[0097] According to the present invention, after the third contact reaction is completed, the obtained reaction solution can be subjected to a sixth post-treatment. Preferably, the sixth post-treatment includes: removing the solvent from the reaction solution by rotary evaporation, diluting (e.g., diluting with an appropriate amount of water), extracting (e.g., extracting with a suitable solvent such as ethyl acetate), drying the obtained organic phase (e.g., drying with a drying agent such as anhydrous sodium sulfate or anhydrous magnesium sulfate), removing the solvent by rotary evaporation, and separating by silica gel column chromatography to obtain the compound shown in formula (d).

[0098] Step 4: The compound shown in formula (d) undergoes a fourth contact reaction in an ozone atmosphere, and zinc powder is added for a fifth contact reaction. After removing the zinc powder, a metal borohydride is added for a sixth contact reaction to obtain a hydroxyl-containing Boc-protected pyrrolidine compound shown in formula (3). In formula (d), the stereoconfiguration of the carbon atoms at positions 2, 3, and 4 is "2R,3R,4R" or "2S,3S,4S", and in formula (3), the stereoconfiguration of the carbon atoms at positions 2, 3, and 4 is "2R,3R,4R" or "2S,3S,4S".

[0099] According to the present invention, in order to ensure sufficient contact between the reaction substrates, the solvent for the fourth contact reaction is selected from one or more of dichloromethane, methanol, ethyl acetate and chloroform, preferably dichloromethane and / or methanol.

[0100] Preferably, the amount of solvent used in the fourth contact reaction is 10-200 mL relative to 1 g of the compound shown in formula (d), preferably 50-100 mL, for example, values ​​such as 50 mL, 70 mL, 90 mL and 100 mL and any range between these values.

[0101] According to the present invention, in order to achieve sufficient contact and reaction with ozone, preferably, the conditions for the fourth contact reaction include: a temperature of -70 to -50°C and a time of 5 to 30 minutes. More preferably, the conditions for the fourth contact reaction include: a temperature of -65°C to -55°C (e.g., values ​​such as -65°C, -60°C, -58°C, and -55°C, and any range thereof), and a time of 10 to 20 minutes (e.g., values ​​such as 10 minutes, 15 minutes, 18 minutes, and 20 minutes, and any range thereof).

[0102] According to the present invention, in order for subsequent reactions to proceed smoothly, preferably, after the fourth contact reaction is completed, a non-reactive gas (e.g., nitrogen, argon, etc.) can be introduced into the reaction system to remove ozone.

[0103] According to the present invention, in order to obtain the ideal intermediate, preferably, the amount of zinc powder used is 0.5-5g, more preferably 1-3g, relative to 1g of the compound shown in formula (d), for example, it can be 1g, 1.5g, 1.85g, 2.5g and 3g and any range between these values.

[0104] According to the present invention, in order to obtain better reaction results, preferably, the conditions for the fifth contact reaction include: a temperature of 10-50°C and a time of 1-8 hours. More preferably, the conditions for the fifth contact reaction include: a temperature of 20-40°C (e.g., values ​​such as 20°C, 25°C, 30°C, and 40°C, or any range thereof), and a time of 2-6 hours (e.g., values ​​such as 2 hours, 3 hours, 4 hours, and 6 hours, or any range thereof). Preferably, the fifth contact reaction is carried out in a non-reactive gas atmosphere, such as nitrogen or argon.

[0105] According to the present invention, in order to obtain the desired compound of formula (3), preferably, the metal borohydride is selected from sodium borohydride or potassium borohydride.

[0106] Preferably, the amount of the metal borohydride used relative to 1g of the compound shown in formula (d) is 0.05-0.5g, more preferably 0.1-0.3g, for example, it can be 0.1g, 0.14g, 0.2g, 0.25g and 0.3g, and any range between these values.

[0107] According to the present invention, in order to improve the reaction yield, preferably, the conditions for the sixth contact reaction include: a temperature of 10-50°C and a time of 20-100 min. More preferably, the conditions for the sixth contact reaction include: a temperature of 20-40°C (e.g., values ​​such as 20°C, 25°C, 30°C, and 40°C, or any range thereof), and a time of 40-80 min (e.g., values ​​such as 40 min, 50 min, 60 min, and 80 min, or any range thereof). Preferably, the sixth contact reaction is carried out in a non-reactive gas atmosphere, such as nitrogen or argon.

[0108] According to the present invention, the fourth contact reaction, the fifth contact reaction and the sixth contact reaction are continuous reactions. The specific process can be as follows: the compound shown in formula (d) is added to the solvent, ozone is introduced to carry out the fourth contact reaction, an inactive gas (such as nitrogen, argon, etc.) is introduced to remove the ozone, zinc powder is added to carry out the fifth contact reaction, the zinc powder is filtered out, and a metal borohydride is added to the filtrate to carry out the sixth contact reaction, so as to obtain the hydroxyl-containing Boc protected pyrrolidine compound shown in formula (3).

[0109] According to the present invention, after the sixth contact reaction is completed, the obtained reaction solution can be subjected to a seventh post-treatment. Preferably, the seventh post-treatment includes: quenching the reaction solution (e.g., quenching with water), extraction (e.g., extraction with a suitable solvent such as dichloromethane), washing the obtained organic phase (e.g., washing with a suitable reagent such as saturated sodium chloride aqueous solution), drying (e.g., drying with a desiccant such as anhydrous sodium sulfate or anhydrous magnesium sulfate), removing the solvent by spin rotation, and separating by silica gel column chromatography to obtain the hydroxyl-containing Boc protected pyrrolidine compound shown in formula (3).

[0110] According to the present invention, the starting material in the preparation method of the above-mentioned hydroxyl-containing Boc protected pyrrolidine compound, namely the compound shown in formula (a), can be obtained commercially or by referring to the references "Cicchi, S.; Hold, I.; Brandi, A." J. Org. Chem. 1993, 58 It was prepared by the method provided in , 5274.”

[0111] The fifth aspect of the present invention provides a glycosidase inhibitor comprising the above-described polyhydroxypyrrolidone ether compounds and their pharmaceutically acceptable salts.

[0112] According to a preferred embodiment of the present invention, the glycosidase inhibitor is used to inhibit one or more of α-glucosidase, β-glucosidase, α-galactosidase, β-galactosidase, α-mannosidase, β-mannosidase, α,α-trehalase, amylase, α-L-rhamnosidase, β-glucuronidase, and β-glucanase, preferably for inhibiting β-glucosidase, more preferably for inhibiting β-glucosidase derived from human lysosomes.

[0113] The sixth aspect of the present invention provides a small molecule drug companion comprising the above-mentioned polyhydroxypyrrolidone ether compounds and their pharmaceutically acceptable salts.

[0114] According to the present invention, the small molecule drug chaperone can be used to enhance the activity of β-glucosidase (GCase) in Gaucher disease fibroblasts. The Gaucher disease fibroblasts can be selected from one or more of N370S, L444P, D409H, G202R, E326K, and T369M fibroblasts, preferably N370S fibroblasts.

[0115] The seventh aspect of the present invention provides the use of the above-mentioned polyhydroxypyrrolidone ether compounds and pharmaceutically acceptable salts thereof in the prevention or treatment of Gaucher disease or Parkinson's syndrome.

[0116] This invention utilizes a special structural design and specific preparation method to obtain a polyhydroxypyrrolidone ether compound and its intermediates that exhibit excellent selectivity and strong inhibitory effect against β-glucosidase. This polyhydroxypyrrolidone ether compound, in particular, demonstrates excellent selectivity and inhibitory ability against β-glucosidase derived from human lysosomes. It also exhibits good molecular chaperone activity against β-glucosidase derived from lysosomes in human N370S fibroblasts, thus making it suitable for the prevention or treatment of Gaucher's disease and Parkinson's syndrome, achieving good preventive or therapeutic effects.

[0117] The present invention will be described in detail below through embodiments.

[0118] In the following examples, the apparatus used is conventional in the art, the operations performed are conventional in the art, and the raw materials and reagents used are commercially available. The compounds shown in formulas (a-1) and (a-2) are referenced in the literature "Cicchi, S.; Hold, I.; Brandi, A." J. Org. Chem. 1993, 58 It was prepared by the method provided in , 5274.”

[0119] Preparation Example 1

[0120] ; (1) Under an argon atmosphere, 4.9 g of the compound shown in formula (a-1) was added to 40 mL of tetrahydrofuran, and 34.4 mL of vinyl magnesium bromide was added at 0 °C. The reaction was carried out at 25 °C for 30 min. After the reaction was completed, the reaction solution was quenched with saturated ammonium chloride aqueous solution, extracted with ethyl acetate, and the obtained organic phase was dried with anhydrous magnesium sulfate and the solvent was removed by rotary evaporation to obtain the compound shown in formula (b-1).

[0121] (2) Under an argon atmosphere, 0.17 g of copper acetate and 10.8 g of zinc powder were added to 50 mL of glacial acetic acid. A glacial acetic acid solution of the compound shown in formula (b-1) (concentration of 0.1 g / mL, the amount of solution used was such that the amount of the compound shown in formula (b-1) was 5 g) was added, and the reaction was carried out at 25 °C for 4 h. After the reaction was completed, the reaction solution was filtered to remove the zinc powder, and the solvent was removed from the filtrate to obtain the compound shown in formula (c-1).

[0122] (3) Under an argon atmosphere, 3 g of the compound shown in formula (c-1) and 3.9 g of sodium bicarbonate were added to 50 mL of tetrahydrofuran, and 3.4 g of ditert-butyl dicarbonate was added at 0 °C. The reaction was carried out at 25 °C for 1 h. After the reaction was completed, the solvent was removed from the reaction solution by rotary evaporation, the product was diluted with water, extracted with ethyl acetate, and the resulting organic phase was dried with anhydrous magnesium sulfate, the solvent was removed by rotary evaporation, and the mixture was separated by silica gel column chromatography to obtain the compound shown in formula (d-1).

[0123] (4) 2.1 g of the compound shown in formula (d-1) was added to 150 mL of dichloromethane, and ozone was introduced at -60 °C for 15 min. After the reaction was completed, argon gas was introduced to remove the ozone, and 3.9 g of zinc powder was added to the reaction solution, and the reaction was carried out at 25 °C for 3 h. After the reaction was completed, the reaction solution was filtered to remove the zinc powder, and 0.3 g of sodium borohydride was added to the filtrate, and the reaction was carried out at 25 °C for 1 h. After the reaction was completed, the reaction solution was quenched with water, extracted with dichloromethane, and the obtained organic phase was washed with saturated sodium chloride aqueous solution, dried with anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation and separated by silica gel column chromatography to obtain the compound shown in formula (3-1).

[0124] Preparation Example 2

[0125] ; According to the method of preparation example 1, the difference is that in step (1), the compound shown in formula (a-1) is replaced with the compound shown in formula (a-2), and the compound shown in formula (3-2) is finally obtained.

[0126] Examples 1-22 This embodiment is used to illustrate the preparation process of the compound shown in formula (2) and the compound shown in formula (1), wherein the substrates and products used are shown in Table 1.

[0127] (1) Under an argon atmosphere, 43 mg of the compound shown in formula (3), 38 mg of sodium hydride and 5 mg of tetrabutylammonium iodide were added to 20 mL of tetrahydrofuran and stirred for 30 min. Then, 120 mg of R was added. 1 X, reacted at 25℃ for 8 h. After the reaction was completed, the reaction solution was quenched with saturated ammonium chloride aqueous solution, diluted with water, extracted with ethyl acetate, and the resulting organic phase was dried with anhydrous magnesium sulfate, the solvent was removed by rotary evaporation, and separated by silica gel column chromatography to obtain the compound shown in formula (2).

[0128] (2) 159 mg of the compound shown in formula (2) was added to 10 mL of methanol, along with 10 mg of Pd / C (Pd content of 10 wt%) and 0.2 mL of hydrochloric acid (concentration of 6 N, solvent of methanol). The reaction was carried out at 25 °C for 8 h under a hydrogen pressure of 1 atm. After the reaction was completed, argon gas was introduced to purge the reaction solution, and the solvent was removed from the filtrate to obtain the hydrochloride salt of the compound shown in formula (1).

[0129] Table 1

[0130] Test Example 1 This test example is used to verify the glycosidase inhibitory activity of the hydrochloride salts of the compounds shown in formulas (1-1) to (1-22).

[0131] (1) Experimental materials: The 4-nitrophenol pyranoside, sodium acetate buffer solution and various glycosidases (α-glucosidase, β-glucosidase, α-galactosidase, β-galactosidase, α-mannosidase, β-mannosidase, α,α-trehalase, amylase, α-L-rhamnosidase, β-glucuronidase, β-glucanase) used in the following tests were all purchased from Sigma-Aldrich.

[0132] (2) Test method: Add 20 μL of 0.1 M sodium acetate buffer solution (pH=4) to a 96-well plate, then add 10 μL of α-glucosidase and 20 μL of hydrochloride aqueous solution of the compound shown in formula (1-1) (added and tested at gradient concentrations of 0.05 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.4 mg / mL, 0.8 mg / mL and 1 mg / mL respectively). Shake the mixture vigorously and incubate at 25 °C for 10 min. Add 100 μL of 1 M sodium carbonate aqueous solution to the 96-well plate to terminate the reaction. Measure the absorbance of the mixture at 400 nm using a microplate reader, calculate the substrate consumption rate, and perform data analysis using the GraFit program (see Leatherbarrow, RJ Grafit 4.0; Eritacus Software: Stains, UK, 1998). The hydrochloride salts of the compounds shown in formulas (1-2) to (1-22) were tested according to the aforementioned method.

[0133] (3) Evaluation results: The results are shown in Table 2. The percentages in Table 2 represent the inhibition rate of the corresponding compound against the glycosidase, indicating its IC50 value against the enzyme. 50 A μM value greater than 1000 indicates low inhibitory activity; while the non-percentages in Table 1 represent the IC50 values ​​of the corresponding compounds for this enzyme. 50 The smaller the (μM) value, the stronger the inhibitory activity.

[0134] Table 2

[0135] Table 2 (Continued)

[0136] Table 2 (Continued)

[0137] As can be seen from Table 2, the polyhydroxypyrrolidone compounds of the present invention have good glycosidase inhibitory activity, especially showing excellent activity against β-glucosidase.

[0138] Test Example 2 This test example uses the compounds shown in formulas (1-7) and (1-8) to evaluate their stability as stabilizers for human lysosomal β-glucosidase.

[0139] (1) Experimental materials: The human lysosomal β-glucosidase, acetate buffer, protein thermal shift dye, DAB (1,4-dideoxy-1,4-imino-D-arabinitol), and IFG (isomulberry leaf alkaloids) used in the following tests were all purchased from Sigma-Aldrich.

[0140] (2) Test method: The melting curve of human lysosomal β-glucosidase was determined by thermal shift analysis using the StepOne real-time polymerase chain reaction (PCR) system from Applied Biosystems, Inc. β-glucosidase (final concentration 0.15 mg / mL) was equilibrated with protein thermal shift dye in acetate buffer (pH=4, concentration 200 mM). Compounds shown in formulas (1-7) or (1-8) (final concentration 10 mM) were added. Samples were aliquoted into MicroAmp Fast 8-tube strips (2 μL per tube) and heated from 25 °C to 99 °C at a heating rate of 1.32 °C / min. The optimal excitation wavelength (490 nm) and emission wavelength (530 nm) for fluorophores were used to detect the protein thermal shift dye. Fluorescence intensity was measured every 0.4 °C. The melting temperature was determined from the first derivative curve using protein thermal shift software v1.3 (Thermo Fisher Scientific). A blank group (NT) and control groups with DAB or IFG were set up according to the aforementioned method.

[0141] (3) Evaluation results: The results are as follows Figure 1 As shown, Figure 1 In the graph, the vertical axis represents the degree of enzyme deformation; a higher value indicates more severe enzyme deformation. The horizontal axis represents the temperature at which the enzyme was subjected to the test. Figure 1 It can be seen that when the compounds of formula (1-7) or formula (1-8) of the present invention are used as stabilizers, the stability of intracellular β-glucosidase can be significantly improved, and the performance is far superior to that of the blank group and the control group. This indicates that the polyhydroxypyrrolidone compounds of the present invention can be used alone or in combination with currently approved enzyme replacement therapy (ERT), and have broad application prospects in the prevention and treatment of Gaucher disease and Parkinson's syndrome.

[0142] Test Example 3 This test example uses the compounds shown in formulas (1-7) and (1-8) to evaluate their performance as small molecule chaperones.

[0143] (1) Experimental materials: Gaucher disease N370S fibroblasts (GM00372) used in the following tests were purchased from Collier Cell Bank, USA. EMEM medium, sodium taurocholate, Triton X-100, citrate buffer (pH=5.2), 4-methylumbelliferone-β-D-glucoside, and fetal bovine serum were all purchased from Sigma-Aldrich.

[0144] (2) Test methods: Gaucher disease N370S fibroblasts were cultured in EMEM medium supplemented with 15 wt% fetal bovine serum. The cells were cultured for 6 days in a water-jacketed incubator at 37°C and 5 vol% CO2. Compounds shown in formulas (1-7) or (1-8) (final concentration 10 mM) were added to the culture system. After washing twice with the culture medium, the cell pellet was homogenized in citrate buffer (pH=5.2) containing 0.25 wt% sodium taurocholate and 0.1 wt% Triton X-100. After centrifugation at 1000g for 5 min, the supernatant was used for enzyme activity determination and protein quantification. β-glucosidase activity in the cells was measured using 4-methylumbelliferone-β-D-glucan as a substrate at pH=5.2. A blank group (NT) and control groups supplemented with DAB or IFG were set up according to the aforementioned method.

[0145] (3) Evaluation results: The results are as follows Figure 2 As shown, Figure 2 In the graph, the vertical axis represents the degree of enhancement of residual enzyme activity in human N370S fibroblasts; the higher the value, the better the effect. (This is achieved through...) Figure 2 It can be seen that when the compounds of formula (1-7) or formula (1-8) of the present invention are used as small molecule chaperones, they can significantly enhance the intracellular human lysosomal β-glucosidase activity in cells present as Gaucher disease N370S, and their molecular chaperone effect is significantly stronger than that of DAB and IFG.

[0146] To visually illustrate the properties and characterization process of the compounds prepared using the method of this invention, the present invention provides, exemplarily, the identification results of some products. See Table 3 for details.

[0147] Table 3

[0148] The preferred embodiments of the present invention have been described in detail above; however, the present invention is not limited thereto. Within the scope of the inventive concept, various simple modifications can be made to the technical solutions of the present invention, including combinations of various technical features in any other suitable manner. These simple modifications and combinations should also be considered as the content disclosed in the present invention and are all within the protection scope of the present invention.

Claims

1. A polyhydroxypyrrolidone ether compound and its pharmaceutically acceptable salt, characterized in that, The structure of this polyhydroxypyrrolidone ether compound is shown in formula (1): Equation (1) ; Among them, R 1 One or more alkyl groups selected from C5-C30; R 2 and R 3 Simultaneously selected from H and one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more alkyl groups and halogens of C1-C12; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (1) is "2R,3R,4R" or "2S,3S,4S".

2. The polyhydroxypyrrolidone ether compound according to claim 1, wherein, R 1 One or more alkyl groups selected from C5-C24; R 2 and R 3 Simultaneously selected from one or more of H and substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more of C1-C6 alkyl groups and halogens; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (1) is "2R,3R,4R" or "2S,3S,4S". Preferably, R 1 Selected from one or more n-alkyl groups from C5-C18; R 2 and R 3 Simultaneously selected from H and one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more of C1-C3 alkyl groups, F, Cl and Br; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (1) is "2R,3R,4R" or "2S,3S,4S". More preferably, R 1 Selected from one or more of n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecanyl, n-hexadecyl, n-heptadecyl, and n-octadecyl; R 2 and R 3 Simultaneously selected from one or more of H, benzyl, p-methylbenzyl, p-ethylbenzyl, p-isopropylbenzyl, p-chlorobenzyl and p-bromobenzyl; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (1) is "2R,3R,4R" or "2S,3S,4S"; More preferably, the polyhydroxypyrrolidone ether compound is selected from one or more compounds shown in the following formula: ； Preferably, the pharmaceutically acceptable salts of the polyhydroxypyrrolidone compounds include one or more of hydrochloride, sulfonate, trifluoroacetate, carbonate, nitrate, phosphate, folic acid, and acetate, and are more preferably one or more of hydrochloride, sulfonate, and trifluoroacetate.

3. A Boc-protected pyrrolidone compound and its pharmaceutically acceptable salt, characterized in that, The structure of the Boc-protected pyrrolidine ether compound is shown in formula (2): Equation (2) ; Among them, R 1 One or more alkyl groups selected from C5-C30, R2 and R3 are both selected from one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more alkyl groups and halogens from C1-C12; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is "2R,3R,4R" or "2S,3S,4S".

4. The Boc-protected pyrrolidone ether compound according to claim 3, wherein, R 1 One or more alkyl groups selected from C5-C24; R2 and R3 are both selected from one or more substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more alkyl groups and halogens from C1-C6; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is "2R,3R,4R" or "2S,3S,4S"; Preferably, R 1 One or more of the C5-C18 n-alkyl groups; R2 and R3 are both selected from one or more of the substituted or unsubstituted benzyl groups, wherein the substituents of the substituted benzyl groups are each independently selected from one or more of the C1-C3 alkyl groups, F, Cl and Br; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is "2R,3R,4R" or "2S,3S,4S"; More preferably, R 1 R2 is selected from one or more of n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecanyl, n-hexadecyl, n-heptadecyl, and n-octadecyl; R2 and R3 are simultaneously selected from one or more of benzyl, p-methylbenzyl, p-ethylbenzyl, p-isopropylbenzyl, p-chlorobenzyl, and p-bromobenzyl; the stereoconfiguration of the carbon atoms at positions 2, 3, and 4 in formula (2) is "2R,3R,4R" or "2S,3S,4S"; More preferably, the Boc-protected pyrrolidone compound is selected from one or more compounds shown in the following formula: ； Preferably, the pharmaceutically acceptable salts of the Boc-protected pyrrolidone compounds include one or more of hydrochloride, sulfonate, trifluoroacetate, carbonate, nitrate, phosphate, folic acid, and acetate, and are more preferably one or more of hydrochloride, sulfonate, and trifluoroacetate.

5. A method for preparing a polyhydroxypyrrolidone ether compound and its pharmaceutically acceptable salt, characterized in that, The method includes: Scenario 1: When R 2 and R 3 When H is present, in the presence of a first catalyst and a first acidic reagent, hydrogen is used to reduce the Boc protected pyrrolidone ether compound shown in formula (2) to obtain the polyhydroxypyrrolidone ether compound shown in formula (1). Scenario 2: When R 2 and R 3 When one or more of the substituted or unsubstituted benzyl groups are present, the Boc-protected pyrrolidone ethers shown in formula (2) are subjected to a deBoc reaction in the presence of a second acidic reagent to obtain the polyhydroxypyrrolidone ethers shown in formula (1). Equation (2) Equation (1) ; Among them, R 1 R 2 R 3 R2 and R3 are as defined in any one of claims 1-4; the stereo configuration of the carbon atoms at positions 2, 3 and 4 in formula (2) is "2R,3R,4R" or "2S,3S,4S"; the stereo configuration of the carbon atoms at positions 2, 3 and 4 in formula (1) is "2R,3R,4R" or "2S,3S,4S".

6. The method according to claim 5, wherein, The first catalyst is selected from one or more of Raney nickel, Pd / C, palladium black, palladium hydroxide, palladium acetate, palladium chloride, platinum chloride, and platinum black, preferably one or more of Raney nickel, Pd / C, palladium hydroxide, and palladium acetate; And / or, relative to 1g of the Boc-protected pyrrolidone compound, the amount of the first catalyst is 0.005-0.5g, preferably 0.01-0.1g; And / or, the first acidic reagent is selected from one or more of hydrochloric acid, sulfuric acid, p-toluenesulfonic acid and trifluoroacetic acid, preferably hydrochloric acid and / or sulfuric acid; And / or, the first acidic reagent is provided in the form of a first acid solution; preferably, the concentration of the first acid solution is 1-10N, more preferably 2-8N; preferably, the amount of the first acid solution used is 0.5-5mL, more preferably 0.8-2mL, relative to 1g of the Boc protected pyrrolidone compound. And / or, the solvent for the reduction treatment is selected from one or more of methanol, ethanol, acetonitrile, dichloromethane, chloroform, tetrahydrofuran, diethyl ether, ethyl acetate, acetic acid, water, 1,4-dioxane and N,N-dimethylformamide, preferably one or more of methanol, ethanol and acetonitrile; And / or, relative to 1g of the Boc protected pyrrolidone compound, the amount of solvent used for the reduction treatment is 10-120mL, preferably 40-80mL; And / or, the conditions for the reduction treatment include: hydrogen pressure of 1-10 atm, temperature of 10-60°C, and time of 2-16 h; more preferably, the conditions for the reduction treatment include: hydrogen pressure of 1-5 atm, temperature of 20-50°C, and time of 5-10 h. And / or, the second acidic reagent is selected from one or more of hydrochloric acid, sulfuric acid, p-toluenesulfonic acid and trifluoroacetic acid, preferably hydrochloric acid and / or sulfuric acid; And / or, the second acidic reagent is provided in the form of a second acid solution; preferably, the concentration of the second acid solution is 1-10N, more preferably 2-8N; preferably, the amount of the second acid solution used is 0.5-5mL, more preferably 0.8-2mL, relative to 1g of the Boc protected pyrrolidone compound; And / or, the solvent for the Boc removal reaction is selected from one or more of methanol, ethanol, acetonitrile, dichloromethane, chloroform, tetrahydrofuran, diethyl ether, ethyl acetate, acetic acid, water, 1,4-dioxane and N,N-dimethylformamide, preferably one or more of methanol, ethanol and acetonitrile; And / or, relative to 1g of the Boc-protected pyrrolidone compound, the amount of solvent used in the Boc removal reaction is 10-120mL, preferably 40-80mL; And / or, the conditions for the Boc removal reaction include: a temperature of 10-60°C and a time of 2-16 h; more preferably, the conditions for the Boc removal reaction include: a temperature of 20-50°C and a time of 5-10 h.

7. A method for preparing a Boc-protected pyrrolidone ether compound and its pharmaceutically acceptable salt, characterized in that, The method includes: in the presence of a second catalyst and a basic reagent, reacting a hydroxyl-containing Boc-protected pyrrolidine compound of formula (3) with R 1 X undergoes an etherification reaction to obtain the Boc protected pyrrolidine ether compound shown in formula (2); Equation (3) Equation (2) ; Among them, R 1 R2 and R3 are as defined in claim 3 or 4, and X is selected from one or more halogens; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (3) is "2R,3R,4R" or "2S,3S,4S"; the stereoconfiguration of the carbon atoms at positions 2, 3 and 4 in formula (2) is "2R,3R,4R" or "2S,3S,4S".

8. The method according to claim 7, wherein, The hydroxyl-containing Boc-protected pyrrolidine compound is selected from one or more compounds shown in the following formula: ； And / or, the second catalyst is selected from one or more of tetrabutylammonium iodide, tetrabutylammonium chloride, tetrabutylammonium bromide, tetrabutylammonium fluoride and crown ether, preferably tetrabutylammonium iodide and / or tetrabutylammonium bromide; And / or, relative to 1g of the hydroxyl-containing Boc-protected pyrrolidine compound, the amount of the second catalyst is 0.01-1g, preferably 0.05-0.5g; And / or, the alkaline reagent is selected from one or more of potassium carbonate, sodium hydride, potassium hydroxide, sodium hydroxide and calcium hydroxide, preferably potassium carbonate and / or sodium hydride; And / or, relative to 1g of the hydroxyl-containing Boc-protected pyrrolidine compound, the amount of the basic reagent is 0.1-5g, preferably 0.5-2g; And / or, relative to 1g of the hydroxyl-containing Boc-protected pyrrolidine compound, R 1 The dosage of X is 0.1-10g, preferably 0.5-5g; And / or, the solvent for the etherification reaction is selected from one or more of diethyl ether, tetrahydrofuran, N,N-dimethylformamide, 1,4-dioxane and dichloromethane, preferably tetrahydrofuran and / or N,N-dimethylformamide; And / or, relative to 1g of the hydroxyl-containing Boc-protected pyrrolidine compound, the amount of solvent used in the etherification reaction is 1-1000mL, preferably 200-600mL; And / or, the conditions for the etherification reaction include: a temperature of 10-50°C and a time of 2-16 h; more preferably, the conditions for the etherification reaction include: a temperature of 20-40°C and a time of 6-12 h.

9. Glycosidase inhibitors comprising the polyhydroxypyrrolidone ether compound of claim 1 or 2 and its pharmaceutically acceptable salt.

10. Small molecule drug companions comprising the polyhydroxypyrrolidone ethers of claim 1 or 2 and their pharmaceutically acceptable salts.

11. The use of the polyhydroxypyrrolidone ether compound of claim 1 or 2 and its pharmaceutically acceptable salt in the prevention or treatment of Gaucher disease or Parkinson's syndrome.

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