CN114478659B

CN114478659B - Preparation method of disaccharide compound

Info

Publication number: CN114478659B
Application number: CN202210389183.5A
Authority: CN
Inventors: 熊美军; 胡明宇; 秦刚
Original assignee: Genequantum Healthcare Suzhou Co Ltd
Current assignee: Genequantum Healthcare Suzhou Co Ltd
Priority date: 2022-04-14
Filing date: 2022-04-14
Publication date: 2022-07-15
Anticipated expiration: 2042-04-14
Also published as: CN114478659A

Abstract

The invention provides a preparation method of a disaccharide compound, an intermediate compound and a preparation method thereof, wherein the method uses cheap and easily-obtained reaction reagents, has simple operation method, higher reaction purity and yield and low process cost, and is suitable for large-scale production.

Description

Preparation method of disaccharide compound

Technical Field

The invention relates to the field of drug synthesis, in particular to a preparation method of a disaccharide compound.

Background

The natural product library has diversified structures and functions, and thus becomes one of the important sources of small molecule drug leads. The carbohydrate is one of important life substances and plays a key role in signal transduction, biological recognition and the like in organisms. Carbohydrates for biological research and commercial applications are often difficult to synthesize by the biogenic route due to high functionalization, and are mostly obtained by chemical synthesis methods. The polyhydroxy structural characteristics of the saccharide enable the synthesis of the saccharide derivatives to have greater challenges, the difficulties mainly comprise the stereoselective construction of glycosidic bonds and the regioselective functionalization of polyhydroxy, and the synthesis of functionalized oligosaccharide molecules can be realized through reasonable route design, a protecting group strategy and the selection of efficient glycosidation conditions.

Disclosure of Invention

The invention provides a preparation method of a compound HX21010 with a disaccharide structure, which comprises the following reaction steps:

taking a compound H0002 and a compound H0003 as reaction raw materials, and preparing a compound HX21010 in an inert atmosphere in an organic solvent 1 under the action of a catalyst 1;

wherein, the organic solvent 1 is selected from any one or any combination of dichloromethane, carbon tetrachloride, chloroform, ethanol, methanol, dimethyl sulfoxide (DMSO), N-Dimethylformamide (DMF) and Tetrahydrofuran (THF); the catalyst 1 is a combination of 1- (phenylsulfinyl) piperidine, 2,4, 6-tri-tert-butyl pyrimidine and trifluoromethanesulfonic anhydride, and the molar ratio of the dosage of the combination is 1-1.5:2-3: 1-2;

the compound H0003 is prepared by the following method:

wherein the alkali is selected from organic alkali or inorganic alkali, preferably sodium carbonate (Na)₂CO₃) Potassium carbonate (K)₂CO₃) Sodium hydroxide (NaOH), potassium hydroxide (KOH), cesium carbonate (Cs)₂CO₃) Any one or any combination of tert-butyl sodium, sodium methoxide, sodium ethoxide, triethylamine, pyridine, morpholine, N-methylmorpholine and N, N-diisopropylethylamine;

wherein, the organic solvent 2 is selected from any one of methanol, ethanol, Tetrahydrofuran (THF), N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), acetonitrile, acetone and dioxane or any combination thereof.

In a preferred embodiment of the present invention, the base is sodium methoxide or sodium ethoxide.

In a preferred embodiment of the present invention, the organic solvent 2 is selected from any one of methanol, ethanol, and Tetrahydrofuran (THF), or any combination thereof.

In a preferred embodiment of the present invention, the organic solvent 2 is a mixed solvent of methanol and Tetrahydrofuran (THF).

In a preferred embodiment of the present invention, compound H0003i is prepared by the following steps:

wherein X represents OH;

wherein, the organic solvent 3 is selected from any one of toluene, acetonitrile, chloroform, dichloromethane, methanol, ethanol, acetone, Tetrahydrofuran (THF), N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dioxane, carbon tetrachloride or any combination thereof.

In a preferred embodiment of the present invention, the catalyst 2 is selected from one of trimethylsilyl trifluoromethanesulfonate, trifluoromethanesulfonic acid, tin tetrachloride, p-toluenesulfonic acid, trifluoroacetic acid, or any combination thereof.

In a preferred embodiment of the present invention, compound H0003 is prepared by the following method:

wherein, X represents OH;

wherein, the organic solvent 3 is selected from any one of toluene, acetonitrile, chloroform, dichloromethane, methanol, ethanol, acetone, Tetrahydrofuran (THF), N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dioxane and carbon tetrachloride or any combination thereof;

wherein, the catalyst 2 is selected from any one of or any combination of trimethylsilyl trifluoromethanesulfonate, trifluoromethanesulfonic acid, stannic chloride, p-toluenesulfonic acid and trifluoroacetic acid;

wherein the alkali is selected from sodium carbonate (Na)₂CO₃) Potassium carbonate (K)₂CO₃) Sodium hydroxide (NaOH), potassium hydroxide (KOH), cesium carbonate (Cs)₂CO₃) Any one or any combination of tert-butyl sodium, sodium methoxide, sodium ethoxide, triethylamine, pyridine, morpholine, N-methylmorpholine and N, N-diisopropylethylamine;

In a preferred embodiment of the present invention, the organic solvent 2 is selected from any one of methanol, ethanol, and Tetrahydrofuran (THF), or a mixed solvent of methanol and Tetrahydrofuran (THF).

In the technical scheme of the invention, the step of preparing the compound HX21010 by using the compound H0002 and the compound H0003 as reaction raw materials is carried out in an inert atmosphere, the reaction temperature is-65 ℃ to 0 ℃, the organic solvent 1 is selected from any one or combination of dichloromethane, carbon tetrachloride, chloroform, ethanol, methanol, dimethyl sulfoxide (DMSO), N-Dimethylformamide (DMF) and Tetrahydrofuran (THF), the catalyst 1 is a combination of 1- (phenylsulfinyl) piperidine, 2,4, 6-tri-tert-butylpyrimidine and trifluoromethanesulfonic anhydride, and the molar ratio of 1- (phenylsulfinyl) piperidine, 2,4, 6-tri-tert-butylpyrimidine and trifluoromethanesulfonic anhydride is 1-1.5:2-3: 1-2.

In a preferred embodiment of the present invention, compound H0002 is prepared by the following method:

step (i): under the protection of inert gas, in an organic solvent and in the presence of a catalyst, carrying out acylation reaction to prepare a compound 2 a;

step (ii): under the action of a catalyst, reacting the compound H0002a prepared in the step (i) with p-methylthiophenol (TolSH) in an organic solvent to prepare an intermediate H0002 b;

step (iii): (iii) preparing intermediate H0002c from intermediate H0002b prepared in step (ii) by deacetylation protection under basic conditions in an organic solvent;

step (iv): (iv) intermediate H0002c prepared by step (iii) in organic solvent in the presence of acidic catalyst with PhCH (OMe)₂Reacting to obtain an intermediate H0002 d;

and (v) preparing the intermediate H0002 by benzylation of the intermediate H0002d prepared in the step (iv) in an organic solvent under the action of a catalyst.

In a preferred embodiment of the present invention, the inert gas in step (i) is nitrogen or argon;

in a preferred embodiment of the present invention, wherein the organic solvent in step (i) is selected from any one of dichloromethane, methanol, ethanol, Tetrahydrofuran (THF), N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO) or any combination thereof;

in a preferred embodiment of the present invention, the catalyst in step (i) is any one selected from triethylamine, pyridine, and 4-dimethylaminopyridine, or any combination thereof;

in a preferred embodiment of the present invention, wherein the catalyst in step (i) is selected from a mixture of triethylamine and 4-dimethylaminopyridine;

in a preferred embodiment of the present invention, wherein, the acylating agent in step (i) is acetyl chloride or acetic anhydride;

in a preferred embodiment of the present invention, the reaction temperature in step (i) is 0 to 25 ℃.

In a preferred embodiment of the present invention, the organic solvent in step (ii) is selected from any one of dichloromethane, methanol, ethanol, Tetrahydrofuran (THF), N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), or any combination thereof;

in a preferred embodiment of the present invention, the catalyst in step (ii) is any one of boron trifluoride diethyl etherate, trifluoroacetic acid and trifluoromethanesulfonic acid, or any combination thereof;

in a preferred embodiment of the present invention, the reaction temperature in step (ii) is 0 to 25 ℃.

In a preferred embodiment of the present invention, the alkaline condition in step (iii) is sodium methoxide, sodium ethoxide, sodium hydroxide (NaOH), sodium carbonate (Na)₂CO₃) Any one or any combination thereof;

in a preferred embodiment of the present invention, the solvent in step (iii) is selected from any one of water, acetonitrile, methanol, ethanol, N-Dimethylformamide (DMF), Tetrahydrofuran (THF), dimethyl sulfoxide (DMSO), dioxane, or any combination thereof;

in a preferred embodiment of the present invention, the solvent in step (iii) is preferably methanol.

In a preferred embodiment of the present invention, the reaction solvent in step (iv) is any one of water, acetonitrile, methanol, ethanol, N-Dimethylformamide (DMF), Tetrahydrofuran (THF), dimethyl sulfoxide (DMSO), dioxane, or any combination thereof;

in a preferred embodiment of the present invention, wherein the reaction solvent in step (iv) is preferably acetonitrile;

in a preferred embodiment of the present invention, wherein step (iv) is carried out under an inert atmosphere (e.g., nitrogen or argon);

in a preferred embodiment of the present invention, wherein the acidic catalyst in step (iv) is selected from any one of sulfuric acid, benzenesulfonic acid, p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid or any combination thereof; preferably benzenesulfonic acid;

in a preferred embodiment of the present invention, the reaction temperature in step (iv) is from room temperature to 50 ℃.

In a preferred embodiment of the present invention, wherein the catalyst in step (v) is selected from tetrabutylammonium iodide, tetrabutylammonium bromide and tetrabutylammonium chloride; preferably tetrabutylammonium iodide;

in a preferred embodiment of the present invention, step (v) is performed under alkaline conditions, wherein the base is selected from the group consisting of sodium hydride (NaH), sodium hydroxide (NaOH), potassium hydroxide (KOH), and sodium carbonate (Na)₂CO₃) Any one or any combination thereof; preferably sodium hydride (NaH);

in a preferred embodiment of the present invention, wherein the benzylation reagent in step (v) is selected from benzyl bromide (BnBr) or benzyl chloride (BnCl);

in a preferred embodiment of the present invention, the organic solvent in step (v) is selected from acetonitrile, methanol, ethanol, N-Dimethylformamide (DMF), Tetrahydrofuran (THF), dimethyl sulfoxide (DMSO), dioxane, preferably Tetrahydrofuran (THF);

in a preferred embodiment of the present invention, the reaction temperature in step (v) is from room temperature to 50 ℃.

Detailed Description

The following detailed description of specific embodiments of the present invention is provided for illustrative purposes only and is not intended to limit the invention.

EXAMPLE 1 preparation of Compound H0002a

To a 500 mL three-necked flask were added D-mannose (18g, 99.89 mmol), DMAP (305mg, 2.5 mmol), triethylamine (101g, 998.9 mmol) and dichloromethane (220 mL). After displacing nitrogen three times, acetic anhydride (61.2g, 599.37 mmol) was added at 0 ℃ and stirred for 10 minutes, the ice-water bath was removed and the reaction solution was stirred at room temperature overnight until TLC (developing reagent: EA/PE = 1/1) showed the end of the reaction. The reaction was quenched with saturated sodium bicarbonate solution, the combined organic phases were extracted with dichloromethane (200 mL × 3), washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (eluent: EA/PE = 1/2) to give H0002a (colorless viscous oil, 37.4 g, yield 95.9%).

EXAMPLE 2 preparation of Compound H0002b

Compound H0002a (37.4 g, 95.8 mmol) obtained in example 1 was taken in a 1000 mL single-neck flask, and p-toluenesulphonol (23.8 g, 191.6 mmol) and anhydrous dichloromethane (350 mL) were added. The mixture was evacuated and purged with nitrogen three times, and boron trifluoride diethyl etherate (20.4 g, 143.7 mmol) was added thereto at 0 ℃. The system was then refluxed overnight until the reaction was substantially complete as detected by TLC. The reaction was then quenched by the slow addition of triethylamine at 0 ℃ and concentrated under reduced pressure to remove the solvent to give the crude product as an oil. The crude product was wet loaded and purified by column chromatography (eluent: EA/PE = 1/3) to yield H0002b (colorless viscous oily liquid, 37.1 g, yield 85.2%).

EXAMPLE 3 preparation of Compound H0002c

Compound H0002b (37.1 g, 81.6 mmol) obtained in example 2 was charged into a 1000 mL two-necked flask, and methanol (450 mL) was added to the flask. Nitrogen was replaced three times, sodium methoxide/methanol solution (1.6 mL, 5M, 8.16 mmol) was added to the reaction, the resulting reaction was stirred at room temperature for 2 hours, and TLC (developing solvent: MeOH/DCM =1/10) indicated the reaction was complete. The pH = 6.5-7 was adjusted by slow addition of 1M hydrochloric acid solution. The solution was concentrated under reduced pressure, toluene was added and water was removed by azeotropic effect to give H0002c as a crude white solid which was used in the next reaction without further purification.

EXAMPLE 4 preparation of Compound H0002d

Example 4-1

All the crude H0002c (81.6 mmol) obtained in example 3 were placed in a 1000 mL single-neck flask at room temperature and successively added p-toluenesulfonic acid monohydrate (3.1 g, 16.3 mmol) and anhydrous acetonitrile (300 mL) and dissolved sufficiently. The nitrogen was replaced by evacuation three times, and benzaldehyde dimethyl acetal (24.8 g, 163.2 mmol) was added under nitrogen protection. The reaction was stirred at room temperature overnight until completion of the reaction as detected by TLC (developing solvent: MeOH// DCM/= 1/10). The reaction was quenched by addition of saturated sodium carbonate solution, the combined organic phases were washed successively with water, saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the resulting crude product was purified by slurrying with acetonitrile to give H0002d (white solid, 24.4 g, combined two-step yield 79.9%).¹H NMR (400 MHz, chloroform-d) delta 7.51-7.39 (m, 7H), 7.20-7.16 (m,2H), 5.62 (s, 1H), 5.55 (s, 1H), 4.39 (td,J = 9.7, 4.9 Hz, 1H), 4.33 (d, J = 3.4 Hz, 1H), 4.27 (dd, J = 10.3, 5.0 Hz, 1H), 4.16 (dd, J = 9.7, 3.4 Hz, 1H), 4.09 – 3.99 (m, 1H), 3.90 – 3.85 (m, 1H), 2.39 (s, 3H)。 C₂₀H₂₂O₅SNa⁺ [M+Na]⁺ms (esi): calculated values: 397.1, actual value: 397.3.

comparative example 1

Referring to the method of example 4-1, the ratio of the reactant H0002c to the benzaldehyde dimethyl acetal was changed to 1:5, and other reaction conditions were not changed, and the final product H0002d was not isolated.

Comparative example 2

Referring to the method of example 4-1, the ratio of the reactant H0002c to benzaldehyde dimethyl acetal was changed to 1:1.5, and other reaction conditions were not changed to obtain the final product H0002d with a yield of 36.1%.

Comparative example 3

Referring to the method of example 4-1, the ratio of the reactant H0002c to benzaldehyde dimethyl acetal was changed to 1:2, and other reaction conditions were unchanged to obtain the final product H0002d with a yield of 75.0%.

Comparative example 4

Referring to the method of example 4-1, the ratio of the reactants H0002c to benzaldehyde dimethyl acetal was 1:2, the reaction solvent was changed to dichloromethane, and other reaction conditions were not changed to obtain the final product H0002d with a yield of 50.3%.

Comparative example 5

Referring to the procedure of example 4-1, the reaction mixture H0002c and benzaldehyde dimethyl acetal were used in a ratio of 1:2, and the type of catalyst was changed to 20mol% of 10-camphorsulfonic acid, and other reaction conditions were unchanged to obtain the final product H0002d in a yield of 31.1%.

Comparative example 6

Referring to the procedure of example 4-1, the reaction mixture H0002c and benzaldehyde dimethyl acetal were used in a ratio of 1:2, and the catalyst type was changed to 20mol% pyridinium p-toluenesulfonate at a reaction temperature of 50 ℃ under otherwise unchanged reaction conditions, to obtain the final product H0002d in a yield of 8.2%.

EXAMPLE 5 preparation of Compound H0002

The compound H0002d (15.4 g, 41.1 mmol) obtained in example 4 and anhydrous tetrahydrofuran (150 mL) were placed in a 500 mL two-necked flask, the nitrogen was replaced by evacuation three times, the system was cooled to 0 deg.C, sodium hydride (4.1 g, 60 wt%, 102.8 mmol) was added in portions, the ice bath was removed and the temperature was raised to room temperature, and the mixture was stirred under nitrogen for 2 hours, followed by addition of tetrabutylammonium iodide (1.5 g, 4.1 mmol) and benzyl bromide (21.1 g, 123.4 mmol). The reaction was stirred at room temperature for 2 hours and the end of the reaction was detected by TLC (EA/PE = 1/8). Slowly adding methanol dropwise to quench the reaction, adding a small amount of water when bubbles are not emitted any more to ensure complete quenching, concentrating to remove tetrahydrofuran, adding ethyl acetate and water to extract and separate liquid, extracting an aqueous phase with ethyl acetate (150 mL × 3), combining the obtained organic phases, washing with common salt water, drying with anhydrous sodium sulfate, filtering and concentrating, and purifying an obtained crude product by column chromatography (EA/PE = 1/10-1/4) to obtain H0002 (light white solid, 22.0 g, yield 96.5%).¹H NMR (400 MHz, chloroform-d) δ 7.63-7.57 (m,2H), 7.50-7.30 (m, 13H), 7.18 (d, J = 7.9 Hz, 3H), 5.72 (s, 1H), 5.52 (d,J = 1.4 Hz, 1H), 4.90 (d, J = 12.2 Hz, 1H), 4.79 (s, 2H), 4.73 (d, J = 12.2 Hz, 1H), 4.42 – 4.34 (m, 2H), 4.30 (dd, J = 10.2, 3.8 Hz, 1H), 4.11 (dd, J = 3.3, 1.5 Hz, 1H), 4.06 (dd, J = 9.5, 3.3 Hz, 1H), 3.95 (t, J = 9.9 Hz, 1H), 2.41 (s, 4H)。C₃₄H₃₅O₅S+ [M+H]⁺ms (esi): calculated values are: 555.2, actual value: 555.4.

EXAMPLE 6 Synthesis of intermediate HX21005a

Glucosamine hydrochloride (10.78 g, 50 mmol) and methanol (200 mL) were added to a 500 mL three-necked flask in this order at room temperature, and sufficiently dissolved. Vacuumizing and replacing nitrogen for three times, and sequentially adding copper sulfate pentahydrate (1.25 g, 5 mmol) and 1H-imidazole-1-sulfonyl azide (12.6 g, 60 mmol) into the system under the protection of nitrogen. The system was then placed in an ice bath, cooled to 0 ℃ and triethylamine (35 mL, 250 mmol) was added slowly. After the addition was complete, the ice-water bath was removed and the reaction solution was stirred at room temperature under nitrogen overnight until the end of the reaction as detected by TLC (developer: MeOH/DCM = 1/2). The reaction solution is filtered by diatomite, washed by dichloromethane and concentrated, and the obtained crude product is subjected to azeotropic concentration by toluene to remove water for three times. The crude brown product (50 mmol) was obtained and used in the next reaction without purification.

The crude product from the previous step (50 mmol) was placed in a 500 mL single-neck flask and 300 mL of anhydrous dichloromethane, 4-dimethylaminopyridine (280 mg, 2.5 mmol) and acetic anhydride (26 mL, 275 mmol) were added sequentially at room temperature. Vacuumizing for replacing nitrogen for three times, placing the system in an ice bath to reduce the temperature to 0 ℃ under the protection of nitrogen, and slowly adding triethylamine (150 mL). After addition, the ice bath was removed and the temperature was raised to room temperature. The reaction was stirred at rt under nitrogen overnight until the end of the reaction as detected by TLC (developing solvent: EtOAc/PE = 1/3). The reaction was quenched by adding a saturated sodium bicarbonate solution to the system, the dichloromethane (500 mL × 3) was used to extract the separated liquid, the combined organic phases were dried over anhydrous sodium sulfate, filtered, the resulting solution was concentrated, wet loaded, and purified by column chromatography (EtOAc/PE = 0-1/3) to give HX21005a (light yellow oily liquid, 11.32 g, α/β = 0.4: 0.6, two steps total yield 60.7%).¹H NMR (400 MHz, chloroform-d) delta 6.32 (d,J = 3.6 Hz, 0.4H, H-1α), 5.59 (d, J = 8.6 Hz, 0.6H, H-1β), 5.47 (dd, J = 10.5, 9.4 Hz, 0.4H), 5.15–5.04 (m, 2H), 4.34–4.29 (m, 1H), 4.16–4.05 (m, 2H), 3.84 (ddd, J = 9.7, 4.5, 2.2 Hz, 0.6H), 3.71–3.67 (m, 1H), 2.21 (s, 3H), 2.12 (s, 1.2H), 2.11 (s, 1.8H), 2.09 (s, 3H), 2.06 (m, 2H), 2.04 (s, 1.8H)。C₁₄H₂₃N₄O₉ ⁺ [M+NH₄]⁺ms (esi): calculated value 391.1, actual value: 391.3.

EXAMPLE 7 Synthesis of intermediate HX21005b

To a 250mL two-necked flask, the compound HX21005a (5.32 g, 14.25 mmol) prepared in example 6 was added and then anhydrous dichloromethane (80 mL) was added to dissolve sufficiently at room temperature. The nitrogen gas was replaced three times, and p-toluenesulfonol (4.4 g, 28.5 mmol) and boron trifluoride diethyl etherate (3.5 mL, 35.63 mmol) were sequentially added to the system under the protection of nitrogen gas. The reaction solution was raised to 50-60 deg.C and stirred under reflux overnight until the TLC detection reaction was substantially complete. At room temperature, triethylamine was slowly added to change the solution from dark brown to light brown. The solution was concentrated, wet loaded, and purified by column chromatography (EtOAc/PE = 0-1/4) to give HX21005b (light brown oil, 4.61 g, α/β = 0.72: 0.28, 74.0% yield).¹H NMR (500 MHz, CDCl₃): δ 7.48 (d, J = 8.0 Hz, Ph-H), 7.39 (d, J = 8.0 Hz, Ph-H), 7.16 (d, J = 8.0 Hz, Ph-H), 7.13 (d, J = 8.0 Hz, Ph-H), 5.57 (d, J _{1α, 2α}= 6.0 Hz, H-1α), 5.54 (dd, J _3α,2α = J _{3α, 4α}= 9.2 Hz, H-3α), 5.07 (dd, J _{3β, 2β}= J _{3β, 4β}= 9.5 Hz, H-3β), 5.04 (dd, J _4α,3α = J _{4α, 5α} = 9.0 Hz, H-4α), 4.90 (dd, J _{4β, 3β}= J _{4β, 5β}= 10.0 Hz, H-4β), 4.62 (ddd, J _5α,4α = 10.0 Hz, J _{5α, 6αa} = 5.0 Hz, J _{5α, 6αa}= 2.0 Hz, H-5α), 4.43 (d, J _{1β, 2β}= 10.0 Hz, H-1β), 4.29 (dd, J _6αa,6αb = 12.0 Hz, J _{6αa, 5α} = 5.0 Hz, H-6αa), 4.23 (dd, J _6βa,6βb = 12.5 Hz, J _{6βa, 5β}= 5.0 Hz, H-6βa), 4.17 (dd, J _6βa,6βb = 12.0 Hz, J _{6βb, 5β}= 2.5 Hz, H-6βb), 4.06 (dd, J _2α,3α = 10.5 Hz, J _{2α, 1α} = 6.0 Hz, H-2α), 4.03 (dd, J _{6αb, 6αa} = 12.0 Hz, J _{6αb, 5α}= 2.0 Hz, H-6αb), 4.62 (ddd, J _5β,4β= 10.0 Hz, J _{5β, 6β} = 5.0 Hz, J _{5β, 6βb}= 2.5 Hz, H-5β), 3.37 (dd, J _{2β, 1β}= J _{2β, 3β}= 10.0 Hz, H-2β), 2.38 (s, Me-β), 2.34 (s, Me-α), 2.10 (s, Ac-α), 2.09 (s, Ac-β), 2.06 (s, Ac-α, Ac-β), 2.04 (s, Ac-α), 2.01 (s, Ac-β)。C₁₉H₂₇N₄O₇S⁺ [M+NH₄]⁺Ms (esi): calculated values: 455.2, actual value: 455.3.

EXAMPLE 8 Synthesis of intermediate HX21005

HX21005b (5.57 g, 12.73 mmol) prepared in example 7 and methanol (50 mL) were added to a 250mL single-neck flask at room temperature and dissolved sufficiently. The system was purged with nitrogen three times under vacuum, sodium methoxide (0.26 mL, 5M in MeOH, 1.273 mmol) was added under nitrogen, stirring at room temperature, and the reaction was followed by TLC (developer: EtOAc/PE = 1/5) and about 1 hour was complete. The solution was neutralized by addition of dilute hydrochloric acid (1M) to pH = 7. After the solution was concentrated, it was azeotropically concentrated with toluene to remove water three times. The crude product was used in the next reaction without purification.

The crude reaction product from the previous step (12.73 mmol) was placed in a 250mL single-neck flask at room temperature, and p-toluenesulfonic acid monohydrate (484 mg, 2.55 mmol) and anhydrous acetonitrile (60 mL) were added in this order and dissolved sufficiently. The nitrogen was replaced by vacuum three times, and benzaldehyde dimethyl acetal (9.5 mL, 63.65 mmol) was added under nitrogen. The reaction was stirred at room temperature overnight until the end of the reaction as detected by TLC (developing solvent: EtOAc/PE = 1/5). The reaction was quenched by adding saturated sodium carbonate solution to the system, the dichloromethane (100 mL × 3) was extracted for liquid separation, the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, wet-loaded, and purified by column chromatography (eluent: EtOAc/PE = 1/10-1/3) to give HX21005 (near white solid, 4.31 g, 84.8% over two steps). The data for the beta-isomer is shown,¹h NMR (400 MHz, chloroform-d) delta 7.56-7.47 (m, 4H), 7.47-7.37 (m, 3H), 7.26-7.18 (m,2H), 5.56 (s, 1H), 4.49(d, J = 10.1 Hz, 1H), 4.41 (dd, J = 10.6, 4.4 Hz, 1H), 3.86–3.70 (m, 2H), 3.53–3.39 (m, 2H), 3.35 (t, J = 9.6 Hz, 1H), 2.94 (s, 1H), 2.42 (s, 3H)。 C₂₀H₂₂N₃O₄S⁺ [M+H]⁺Ms (esi): calculated value 400.1, actual value: 399.9. the data for the alpha isomer is shown,¹h NMR (400 MHz, chloroform-d) δ 7.63–7.54 (m, 2H), 7.52–7.39 (m, 5H), 7.25–7.14 (m, 2H), 5.60 (s, 1H), 5.53 (d, J = 5.4 Hz, 1H), 4.52–4.39 (m, 1H), 4.28 (dd, J = 10.4, 4.9 Hz, 1H), 4.07 (t, J = 9.5 Hz, 1H), 3.92 (dd, J = 10.0, 5.6 Hz, 1H), 3.79 (t, J = 10.3 Hz, 1H), 3.60 (t, J = 9.3 Hz, 1H), 3.08 (s, 1H), 2.40 (s, 3H)。C₂₀H₂₂N₃O₄S⁺ [M+H]⁺Ms (esi): calculated values are: 400.1, actual value: 399.9.

EXAMPLE 9 Synthesis of intermediate HX21006a

Into a 250mL single-neck flask were sequentially added HX21005 (4.31 g, 10.79 mmol) prepared in example 8 and anhydrous tetrahydrofuran (60 mL) and fully dissolved, the system was evacuated to replace nitrogen three times, then placed in an ice bath and cooled to 0 ℃ and added sodium hydride (518 mg, 60 wt% active ingredient, 12.95 mmol), after the addition was completed, the ice bath was removed and the mixture was warmed to room temperature and stirred, and after 1 hour, tetrabutylammonium iodide (399 mg, 1.08 mmol) and benzyl bromide (2.77 g, 16.18 mmol) were added. The reaction was stirred at room temperature for 2h, followed by TLC (developer: EtOAc/PE = 1/8) to check the reaction, quenched with water after the reaction was complete, extracted with ethyl acetate (100 mL. times.3) and the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, dry-mixed and purified by column chromatography (eluent: EtOAc/PE = 1/20-1/4) to afford HX21006a (pale white solid, 5.14 g, 97.3% yield). The data for the alpha isomer is shown,¹h NMR (400 MHz, chloroform-d) δ 7.63–7.55 (m, 2H), 7.53–7.33 (m, 10H), 7.25–7.17 (m, 2H), 5.67 (s, 1H), 5.56 (d, J = 4.6 Hz, 1H), 5.05 (d, J = 10.9 Hz, 1H), 4.90 (d, J = 10.9 Hz, 1H), 4.59–4.47 (m, 1H), 4.31 (dd, J = 10.4, 4.9 Hz, 1H), 4.10–3.97 (m, 2H), 3.90–3.77 (m, 2H), 2.41 (s, 3H)。

EXAMPLE 10 Synthesis of intermediate H0003f

A100 mL two-necked flask was baked for 5 minutes with a hot air gun under vacuum from an oil pump. After cooling, the activated molecular sieve was added and baked for an additional 5 minutes. After replacing nitrogen by evacuation three times, HX21006a (3.0 g, 6.13 mmol) obtained in example 9 and dry dichloromethane (40 mL) were added to the flask in this order under nitrogen protection, and the mixture was sufficiently dissolved by stirring for 5 minutes, and triethylsilane (9.8 mL, 61.3 mmol) was added thereto and stirred at room temperature for 20 minutes. The reaction was placed in a dry ice acetonitrile bath down to-20 ℃, trifluoroacetic acid (4.5 mL, 60.07 mmol) was slowly added, the reaction stirred at-20 ℃ for 3 hours, TLC (developing agent: EtOAc/PE = 1/8) followed the reaction, after the reaction was complete (ca 3H), saturated sodium bicarbonate solution was added to quench the reaction, dichloromethane (100 mL × 3) was extracted for liquid separation, the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, wet loaded, purified by column chromatography (eluent: EtOAc/PE = 1/10-1/4) to give H0003e (light yellow solid, 2.56 g, yield 85.0%). C₂₇H₃₃N₄O₄S⁺ [M+NH₄]⁺Ms (esi): calculated values are: 509.2, actual value: 509.7.

to a 100mL single-neck flask, H003e (2.56 g, 5.21 mmol), anhydrous dichloromethane (30 mL), 4-dimethylaminopyridine (29 mg, 0.261 mmol), and acetic anhydride (1 mL, 10.42 mmol) were added in that order under a nitrogen atmosphere. 0^oTriethylamine (4.4 mL, 31.26 mmol) was added dropwise at C, followed by stirring overnight at room temperature until the reaction was complete, followed by TLC (developer: EtOAc/PE = 1/8). Adding saturated sodium bicarbonate solution into the system to quench the reaction, IIThe chloromethane (50 mL × 3) extract was separated, the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, dry loaded, and purified by column chromatography (eluent: EtOAc/PE = 1/10-1/4) to give H0003f (near white solid, 2.59 g, yield 93.1%). The data for the alpha isomer is shown,¹h NMR (400 MHz, chloroform-d) δ 7.50 – 7.45 (m, 2H), 7.43 – 7.32 (m, 10H), 7.14 – 7.06 (m, 2H), 5.57 (d, J = 5.4 Hz, 1H), 5.16 (dd, J = 10.2, 9.1 Hz, 1H), 4.91 (d, J = 11.2 Hz, 1H), 4.70 (d, J = 11.2 Hz, 1H), 4.58 – 4.49 (m, 3H), 4.02 (dd, J = 10.2, 5.4 Hz, 1H), 3.85 (dd, J = 10.2, 9.0 Hz, 1H), 3.61– 3.51 (m, 2H), 2.35 (s, 3H), 1.93 (s, 3H)。C₂₉H₃₅N₄O₅S⁺ [M+NH₄]⁺Ms (esi): calculated value 551.2, actual value: 551.4.

EXAMPLE 11 Synthesis of intermediate H0003H

To a 100mL single vial were added in sequence the compound H0003f prepared in example 10 (2.59 g, 4.85 mmol), acetone (30 mL), dissolved well, the system was cooled to 0 ℃, NBS (1.2 g, 6.79 mmol) was added and the temperature was maintained for reaction for 1 hour, then saturated sodium thiosulfate solution was added to the system to quench the reaction, the acetone was concentrated to remove, water (5 mL) was then added, the separated layer was extracted with dichloromethane (50 mL × 3), the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, dry loaded, and purified by column chromatography (eluent: EtOAc/PE = 1/10-1/4) to give H0003g (near white solid, 2.03g, yield 97.9%).

Under a nitrogen atmosphere, H0003g (9.24 g, 21.62 mmol) dry dichloromethane (120 mL), DBU (329 mg, 2.16 mmol) and trichloroacetonitrile (8.7 mL, 86.5 mmol) were added sequentially to a 250mL single-neck flask and the temperature was maintained for 3H at 0 ℃. Concentrating to remove solvent, directly wet loading, and purifying by column chromatography (eluent: EtOAc/PE = 1/15-1/10)) H0003H (light yellow oily liquid, 10.33 g, yield 83.5%, α/β = 1.68: 1) was obtained. The data for the beta-isomer are shown,¹h NMR (400 MHz, chloroform-d) δ 8.82 (s, 1H), 7.40–7.33 (m, 10H), 5.71 (d, J = 8.5 Hz, 1H), 5.15 (t, J = 9.6 Hz, 1H), 4.88 (d, J = 11.4 Hz, 1H), 4.72 (d, J = 11.5 Hz, 1H), 4.56 (s, 2H), 3.79 (dd, J = 9.8, 8.5 Hz, 1H), 3.76-3.72 (m, 1H), 3.62–3.49 (m, 3H), 1.90 (s, 3H)。

EXAMPLE 12 Synthesis of intermediate H0003i

A100 mL Schlenk flask was vacuum-charged with an oil pump, baked for 5 minutes with a hot air gun, cooled, charged with an activated molecular sieve, baked for 5 minutes again, vacuum-charged with nitrogen gas, and then, the compound H0003H (10.2 g, 17.84 mmol) obtained in example 11, anhydrous toluene (50 mL), benzyl alcohol (7.4 mL, 71.4 mmol) were added in this order to dissolve them sufficiently, followed by stirring for 20 minutes. The reaction mixture was cooled to-40 ℃ and trimethylsilyl trifluoromethanesulfonate (0.99 mL, 5.35 mmol) was added slowly and stirred at this temperature until TLC (developing solvent: EtOAc/PE = 1/8) indicated the end of the reaction (about 4 hours). The reaction was quenched by adding a saturated sodium bicarbonate solution to the system, and the separated liquid was extracted with ethyl acetate (100 mL × 3), the combined organic phases were washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, dry-loaded, and purified by column chromatography (eluent: EtOAc/PE = 1/15-1/10) to give H0003i (colorless transparent oily liquid, 8.02g, yield 86.9%).¹H NMR (400 MHz, chloroform-d) delta 7.46-7.31 (m, 15H), 5.04 (t,J = 9.6 Hz, 1H), 5.00 (d, J= 11.8 Hz, 1H), 4.86 (d, J = 11.4 Hz, 1H), 4.76 (d, J = 11.8 Hz, 1H), 4.66 (d, J = 11.4 Hz, 1H), 4.59 (s, 2H), 4.42 (d, J = 8.0 Hz, 1H), 3.64–3.53 (m, 4H), 3.44 (t, J = 9.8 Hz, 1H), 1.89 (s, 3H)。C₂₉H₃₅N₄O₆ ⁺ [M+NH₄]⁺MS (2)(ESI): calculated values: 535.3, actual value: 535.6.

EXAMPLE 13 Synthesis of Compound H0003

Compound H0003i (8.02 g, 15.5 mmol) prepared in example 12, anhydrous tetrahydrofuran (35 mL) and anhydrous methanol (35 mL) were sequentially added to a 250mL single-neck flask at room temperature to dissolve, nitrogen was replaced three times, sodium methoxide (1.55 mL, 5M in MeOH, 7.75 mmol) was added, the reaction mixture was stirred at room temperature overnight, the reaction was detected by TLC (developing solvent: EtOAc/PE = 1/8), and after completion of the reaction, 1M hydrochloric acid was added to neutralize the reaction solution and adjust the reaction solution to neutral. Concentration removed the solvent, concentration by toluene azeotropy to remove water, wet loading, and column chromatography purification (eluent: EtOAc/PE = 1/12-1/8) to afford H0003 (colorless oily liquid, 6.98g, 94.7% yield).¹H NMR (400 MHz, chloroform-d) delta 7.45-7.34 (m, 15H), 4.99 (d,J = 11.9 Hz, 1H ), 4.96 (d, J = 11.3 Hz, 1H), 4.81 (d, J = 11.3 Hz, 1H), 4.73 (d, J = 11.9 Hz, 1H), 4.66 (d, J = 12.0 Hz, 1H), 4.61 (d, J = 12.0 Hz, 1H), 4.40 (d, J = 8.1 Hz, 1H), 3.79 (m, 2H), 3.69 (dd, J = 9.0, 9.2 Hz, 1H), 3.53–3.41 (m, 2H), 3.29 (dd, J = 8.9, 9.5 Hz, 1H), 2.70 (s, 1H)。C₂₇H₃₃N₄O₅ ⁺ [M+NH₄]⁺ms (esi): calculated values: 493.2, actual value: 493.5.

example 14 procedure for preparation of compound HX21010 with Compounds H0002 and H0003

A100 mL Schlenk reaction flask was baked with a heat gun for 5 minutes under vacuum conditions of an oil pump, and after cooling, the activated molecular sieve was added to the system and baked for another 5 minutes. The nitrogen was replaced by evacuation three times, and H0002 (4.41 g, 7.96 mmol) was added to the system under nitrogen protection, followed by stirring for 3 minutes, followed by addition of anhydrous dichloromethane (30 mL) and stirring for 0.5 hour. In the same manner as above, H0003 (1.89 g, 3.98 mmol in 20mL of anhydrous dichloromethane) was added to another 50mL Schlenk reaction flask to which the activated molecular sieve had been added, and the mixture was stirred for 1 hour to predry the mixture to remove residual water in the system.

To the above dried H0002 solution was added 1- (phenylsulfinyl) piperidine (BSP, 1.37g, 6.56 mmol) and 2,4, 6-tri-tert-butylpyrimidine (TTBP, 2.94 g, 11.94 mmol) under nitrogen at room temperature, and the mixture was stirred for 20 minutes. The reaction vial was placed in a dry ice/ethyl acetate bath, cooled to-65 ℃ and trifluoromethanesulfonic anhydride (1.2 mL, 7.16 mmol) was added to the system. After 2 minutes, a solution of H0003 predried previously in dichloromethane was added to the system and the resulting reaction mixture was stirred at-65 ℃ until the end of the reaction (ca. 3H) as detected by TLC (developer: EtOAc/PE = 1/8). The reaction was quenched by addition of saturated sodium bicarbonate solution, and the combined organic phases were washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, wet-loaded, and separated by column chromatography (eluent: EtOAc/PE = 1/12-1/10) to give HX21010 (colorless viscous oily liquid, 2.72 g, yield 75.4%).¹H NMR (400 MHz, chloroform-d) delta 7.55 (dd,J = 7.6, 2.1 Hz, 2H), 7.51 – 7.28 (m, 28H), 5.60 (s, 1H), 5.14 (d, J = 10.4 Hz, 1H), 5.00 (d, J = 12.0 Hz, 1H), 4.94 (d, J = 11.8 Hz, 1H), 4.90 – 4.80 (m, 2H), 4.76 (d, J = 12.0 Hz, 1H), 4.74 – 4.63 (m, 3H), 4.57 (s, 1H, H^1’), 4.48 (d, J = 12.1 Hz, 1H), 4.37 (d, J = 8.1 Hz, 1H，H¹), 4.22 – 4.10 (m, 2H), 4.05 (t, J = 9.3 Hz, 1H), 3.80 (d, J = 3.1 Hz, 1H), 3.72 (dd, J = 11.2, 2.2 Hz, 1H), 3.67 – 3.52 (m, 3H), 3.49 (dd, J = 9.8, 3.1 Hz, 1H), 3.41 (t, J = 9.3 Hz, 1H), 3.35 (dt, J = 9.8, 2.9 Hz, 1H), 3.16 (td, J = 9.7, 4.8 Hz, 1H)。C₅₄H₅₆N₃O₁₀ ⁺ [M+H]⁺ms (esi): calculated values are: 9064, actual value: 906.7.

although the present disclosure has been described above, the scope of the present disclosure is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present disclosure, and such changes and modifications will fall within the scope of the present invention.

Claims

1. A preparation method of a compound HX21010 with a disaccharide structure is characterized by comprising the following steps: the method comprises the following reaction steps:

wherein, the organic solvent 1 is selected from any one or any combination of dichloromethane, carbon tetrachloride, chloroform, ethanol, methanol, dimethyl sulfoxide, N-dimethylformamide and tetrahydrofuran;

the catalyst 1 is a combination of 1- (phenylsulfinyl) piperidine, 2,4, 6-tri-tert-butyl pyrimidine and trifluoromethanesulfonic anhydride, and the molar ratio of 1- (phenylsulfinyl) piperidine, 2,4, 6-tri-tert-butyl pyrimidine to trifluoromethanesulfonic anhydride is 1-1.5:2-3: 1-2;

the compound H0003 is prepared by the following method:

wherein the alkali is selected from any one or any combination of sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, tert-butyl sodium, sodium methoxide, sodium ethoxide, triethylamine, pyridine, morpholine, N-methylmorpholine and N, N-diisopropylethylamine;

wherein, the organic solvent 2 is selected from any one or any combination of methanol, ethanol, tetrahydrofuran, N-dimethylformamide, dimethyl sulfoxide, acetonitrile, acetone and dioxane;

the compound H0003i is prepared by the following method:

wherein X represents OH;

wherein, the organic solvent 3 is selected from any one or any combination of toluene, acetonitrile, chloroform, dichloromethane, methanol, ethanol, acetone, tetrahydrofuran, N-dimethylformamide, dimethyl sulfoxide, dioxane and carbon tetrachloride; the catalyst 2 is selected from one or any combination of trimethylsilyl trifluoromethanesulfonate, trifluoromethanesulfonic acid, tin tetrachloride, p-toluenesulfonic acid and trifluoroacetic acid.

2. The process according to claim 1, wherein the base is sodium methoxide or sodium ethoxide.

3. The preparation method according to claim 1, wherein the organic solvent 2 is selected from any one of methanol, ethanol, tetrahydrofuran, or any combination thereof.

4. The process according to claim 3, wherein the organic solvent 2 is a mixed solvent of methanol and tetrahydrofuran.