CN108774274B

CN108774274B - Sialylmethyl ester methyl glycoside derivative and synthesis method and application thereof

Info

Publication number: CN108774274B
Application number: CN201810474379.8A
Authority: CN
Inventors: 叶新山; 刘颖果; 熊德彩; 蒲小平; 李夏溪
Original assignee: Peking University
Current assignee: Peking University
Priority date: 2015-03-06
Filing date: 2015-03-06
Publication date: 2020-11-03
Anticipated expiration: 2035-03-06
Also published as: CN105985389B; CN108752212B; CN108752212A; CN105985389A; CN108774274A

Abstract

The invention discloses a sialic acid methyl ester methyl glycoside derivative, a synthetic method and application thereof. The invention utilizes the design principle of drugs such as biological electron isosteres and the like to modify acteoside to obtain a series of phenylethanoid glycoside derivatives, sialic acid methyl ester derivatives and 9-decarboxylation rosmarinic acid compounds with neuroprotective activity. The invention further provides a synthesis method of the sialic acid methyl ester methyl glycoside derivative. The neuropharmacology experimental research finds that the sialic acid methyl ester methyl glycoside derivative disclosed by the invention increases the cell survival rate, reduces the apoptosis rate, inhibits the generation of active oxygen in cells, obviously relieves the cell damage and plays a role in neuroprotection through anti-apoptosis, anti-oxidation damage and other ways. The sialic acid methyl ester methyl glycoside derivative provided by the invention has important application potential in preventing or treating neurodegenerative diseases including neurodegenerative diseases, cerebral ischemia and the like.

Description

Sialylmethyl ester methyl glycoside derivative and synthesis method and application thereof

The application is a divisional application of an invention patent application with the title of 'phenylethanoid glycoside analogue and a synthesis method and application thereof', the application number of which is '201510101527.8', and the application date of which is '2015, 3 and 6'.

Technical Field

The invention relates to a sialic acid methyl ester methyl glycoside derivative, and further relates to a synthesis method of the derivative and application of the derivative in preventing or treating nervous system diseases, belonging to the field of sialic acid methyl ester methyl glycoside derivatives, and synthesis methods and application of the derivatives.

Background

Neuronal cell damage or death caused by oxidative stress and free radical damage is a common pathological process in many neurological diseases (e.g., neurodegenerative diseases, ischemic stroke, etc.) (Neurologia,17(2002) 399-. Neurodegenerative diseases are characterized by gradual loss of neuronal structure and function, including neuronal death, etc. (adv. neuron., 69(1996) 153-. Many degenerative diseases including Parkinson (PD), Alzheimer's Disease (AD), Huntington (HD), etc. develop in this way. Cerebral ischemia is another way to cause nerve damage. Ischemia causes cerebral hypoxia, which in turn leads to death of brain tissue through complex, progressive, lethal oxidative stress and free radical damage (biophys. acta,1802(2010) 80-91.). Since the pathological processes of these neurological diseases are not well understood, these diseases are not currently radical cure. Most of the existing medicines are old medicines and new medicines, and the existing medicines can only relieve the symptoms of nerve injury. The most common therapeutic strategy for treating this disease is the use of antioxidants and free radical scavengers.

In the past years, the inventor's topic group and other topic groups found that Phenylethanoid Glycosides (PGs) as novel chemical entities have neuroprotective activity, such as acteoside (Life Sci.,79(2006) 709-716), echinancoside (Eur. J. Pharmacol.,564(2007) 66-74; J. Ethnopharmacol.,97 (2005)) 59-63), and calcoelarioside A (biochem. Pharmacol.,51(1996) 687-691; Planta Med.,66(2000) 746-748), and the like. Phenylethanoid glycosides are present in dicotyledonous plants and are a class of natural glycosides containing (hydroxy, methoxy) substituted phenethyl and (hydroxy, methoxy) substituted cinnamyl, usually with a central glucose as the nucleus, and containing ester and oxygen-glycoside bonds (curr. med. chem.,15(2008) 2592-2613.). The phenylethanoid glycosides compounds have wide pharmacological activities related to neurodegenerative diseases and ischemic stroke, and also have good antioxidant and free radical scavenging activities (Planta Med.,68(2002) 966-one 970.). These properties make phenylethanoid glycosides promising as new candidates against nerve damage.

Nevertheless, the following problems still remain to be solved in the phenylethanoid glycosides: firstly, although the phenylethanoid glycosides compounds are widely distributed in the plant world, the content is low (0.02% -0.40%), and the sample obtained by difficult separation and extraction has very small amount, so that the requirement of biological research cannot be met; secondly, the phenylethanoid glycosides have complex structures, so that the synthesis and research of the phenylethanoid glycosides are very difficult. In addition, the good hydrophilicity of the phenylethanoid glycoside leads to poor membrane permeability, which influences the druggability of the phenylethanoid glycoside. Therefore, the research on the synthesis of the phenylethanoid glycosides compounds and the structural modification of the phenylethanoid glycosides compounds can enhance the neuroprotective activities and the drug forming properties of the phenylethanoid glycosides compounds, such as oxidation resistance, free radical scavenging, apoptosis resistance and the like, and has important significance for the application of the phenylethanoid glycosides compounds in the clinical treatment of nervous system diseases.

Disclosure of Invention

One of the purposes of the invention is to provide a kind of phenylethanoid glycoside derivative, sialic acid methyl ester derivative and 9-decarboxylation rosmarinic acid compound with neuroprotective activity;

the second purpose of the invention is to provide a method for synthesizing the phenylethanoid glycoside derivative, the sialic acid methyl ester derivative and the 9-decarboxylation rosmarinic acid compound;

the third purpose of the invention is to apply the phenylethanoid glycoside derivative, the sialic acid methyl ester derivative and the 9-decarboxylation rosmarinic acid compound to prevent or treat nervous system diseases.

The above object of the present invention is achieved by the following technical solutions:

a phenylethanoid glycoside derivative is a compound shown as a formula (I) or a formula (II):

in the formula: r₁Any one selected from aromatic carboxylic acids; preferably, R₁Is 3, 4-dihydroxycinnamoyl, 3-hydroxy-4-methoxycinnamoyl, 4-chlorocinnamoyl, 3, 4-difluorocinnamoyl, 3, 4-dimethoxycinnamoyl, cinnamoyl, 4-hydroxycinnamoyl or 3, 4-dihydroxyphenylacetyl.

The invention also provides a sialic acid methyl ester methyl glycoside derivative which is a compound shown as a formula (III):

wherein R is₁Any one selected from aromatic carboxylic acids; preferably, R₁Is 3, 4-dihydroxy cinnamoyl.

The invention further provides a 9-decarboxylated rosmarinic acid analogue which is a compound shown as a formula (IV):

The invention provides a method for synthesizing a compound shown in a formula (I), which comprises the following steps:

(1) synthesis of compound 16: (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-6-O-allyloxycarbonyl-D-glucopyranoside;

(2) synthesizing a compound shown as a formula (V) or a formula (VI):

wherein R is¹＝R²＝OAllyl；R¹＝R²＝F；R¹＝R²OMe or R¹＝Cl，R²＝H；

(3) Carrying out coupling reaction on a compound (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside and a compound shown as a formula (V) or a formula (VI) to obtain a compound shown as a formula (VIII) or a formula (IX):

R¹＝R²＝OAllyl，R³＝Ac；R¹＝R²＝F，R³＝Ac；R¹＝R²＝OMe，R³＝Ac；R¹＝Cl，R²＝H，R³＝Ac；

wherein R ═ Ac;

wherein the coupling reaction comprises: dissolving a compound (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside and a compound shown as a formula (V) or a formula (VI) in dichloromethane, and reacting under the condition of DCC and DMAP;

(4) dissolving the reaction product shown in the formula (VIII) or the formula (IX) in the step (3) in a mixed solvent (dichloromethane: methanol ═ 1:1), and reacting in the presence of acetyl chloride to obtain a reaction product shown in the formula (X) or the formula (XI);

wherein R is¹＝R²＝OAllyl，R³＝H；R¹＝R²＝F，R³＝H；R¹＝R²＝OMe，R³＝H；R¹＝Cl，R²＝H，R³＝H；

Wherein R ═ H;

(5) dissolving the reaction product of formula (X) or formula (XI) in step (4) in a mixed solvent (dichloromethane: methanol: 20:1) under Pd/C and toluene sulfonic acid or HClO₄Reacting in the presence of a catalyst to obtain the compound shown in the formula (I).

Wherein, (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside (compound 16) can be prepared by the following method:

(1) condensing the

compound

13 and 3, 4-diallyloxyphenethyl alcohol (compound 9a) to obtain (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-4, 6-O-benzylidene-beta-D-glucopyranoside (compound 14);

(2) debenzylating (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-4, 6-O-benzylidene- β -D-glucopyranoside to give (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl- β -D-glucopyranoside (Compound 15);

(3) the (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-beta-D-glucopyranoside (compound)15) In AocOBt, Et₃Reacting in the presence of N to obtain (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside.

The invention also provides a method for synthesizing the compound of the formula (II), which comprises the following steps:

(1) synthesis of the compound (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-. beta. -D-glucopyranoside (Compound 15):

(2) deacetylating the compound (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-beta-D-glucopyranoside to obtain the compound of formula (II).

Wherein, the compound (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-beta-D-glucopyranoside can be prepared by the following method:

(1) condensing compound 13 with 3, 4-diallyloxyphenethyl alcohol to give (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-4, 6-O-benzylidene- β -D-glucopyranoside (compound 14):

(2) the (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-4, 6-O-benzylidene-beta-D-glucopyranoside is debenzylated to obtain the compound (3, 4-diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-beta-D-glucopyranoside.

The invention also provides a method for preparing the compound shown in the formula III, which comprises the following steps:

dissolving a

compound

21 and 3, 4-dihydroxycinnamic acid in DMF, and carrying out condensation reaction in the presence of PyBOP and NMM to obtain the compound;

the invention further provides a method for synthesizing a compound shown as a formula (IV), which comprises the following steps:

(1) carrying out condensation reaction on 3, 4-diacetyl cinnamic acid and 3, 4-diacetyl phenethyl alcohol to obtain 9' -decarboxylation tetraacetyl rosmarinic acid;

(2) and (3) deacetylating and protecting 9' -decarboxylated tetraacetyl rosmarinic acid to obtain the product.

Preferably, the condensation reaction in step (1) is carried out under the following conditions: (a) adding the compound 3, 4-diacetyl cinnamic acid (compound 11b) and oxalyl chloride into DCM for reacting at room temperature (20-25 ℃); (b) the reaction product was concentrated under Ar protection and 3, 4-diacetylphenethyl alcohol (Compound 9b), DCM and Et were added₃And N, stirring and reacting at room temperature (20-25 ℃) to obtain the catalyst.

The invention utilizes the design principle of medicines such as biological electron isosteres to reform acteoside, and uses (2R,3R,4S,5S,6R) -2- (3, 4-dihydroxyphenylethoxy) -6- (hydroxymethyl) tetrahydro-2-hydro-pyran-3, 4, 5-triol, (2S,4S,5R,6R) -methyl 5-amino-4-dihydroxy-2-methoxy-6- ((1R,2R) -1,2, 3-trihydroxypropyl) tetrahydroxy-2-hydro-pyran-2-carboxylate, 3, 4-dihydroxy phenylethanol is used as a monomer and is connected with organic acid or organic acyl chloride to obtain phenylethanoid glycoside derivatives, sialic acid methyl ester derivatives and 9-decarboxylation rosmarinic acid compounds with neuroprotective activity; the research of neuro-pharmacology experiments shows that the compound provided by the invention can obviously increase the cell survival rate, reduce the apoptosis rate and inhibit the generation of active oxygen in cells, has better activities of resisting oxidation and apoptosis and eliminating free radicals, and has no obvious toxic or side effect. Wherein, the compounds (3, 4-dihydroxyphenylethyl) 4-O- (3, 4-hydroxyphenylethyl) -beta-D-glucopyranoside (compound 5), 9' -decarboxylated rosmarinic acid (compound 6) and 2-O-methyl-5-O- (p-hydroxycinnamoyl) -beta-D-sialoside (compound 8) can obviously lighten the damage of 6-OHDA to cells in a dose-dependent mode through anti-apoptosis, anti-oxidative damage and other ways, and play a role in neuroprotection, and are obviously superior to the Edaravone (Edaravone) which is a medicament on the market; the compound provided by the invention has the neuroprotective effects of resisting oxidation, resisting apoptosis of nerve cells, eliminating free radicals and the like, and can be used for preventing and treating neurodegenerative diseases, cerebral ischemia and other neurological diseases.

Therefore, the invention also provides a pharmaceutical composition for preventing and treating neurodegenerative diseases, cerebral ischemia and other neurological diseases, which comprises an effective amount of phenylethanoid glycoside derivatives, methyl sialyl-methyl ester derivatives or 9-decarboxylated rosmarinic acid compounds and pharmaceutically acceptable carriers or auxiliary materials; the pharmaceutical composition with the neuroprotective effects of oxidation resistance, nerve cell apoptosis resistance and the like can be obtained by combining an effective amount of the phenylethanoid glycoside derivative, the sialic acid methyl ester derivative or the 9-decarboxylated rosmarinic acid compound with a pharmaceutically acceptable carrier or auxiliary material.

After various auxiliary materials and pharmaceutically acceptable excipients or carriers required for preparing different dosage forms are added into the pharmaceutical composition, the pharmaceutical composition is prepared into any clinically acceptable appropriate preparation by a conventional pharmaceutical preparation method, for example, the pharmaceutical composition can be an injection preparation (powder injection, freeze-dried powder injection, water injection, infusion and the like), tablets, oral liquid, granules, capsules, soft capsules, dropping pills and the like; wherein, the auxiliary materials can be antioxidant complexing agent, filling agent, framework material and the like; the pharmaceutically acceptable carrier is one or more of xylitol, mannitol, lactose, fructose, dextran, glucose, polyvinylpyrrolidone, low molecular dextran, sodium chloride, calcium gluconate or calcium phosphate.

Abbreviation list

The invention disclosed herein uses the following chemical nomenclature:

DMF N, N-dimethylformamide

IPA isopropyl alcohol

NMM 4-methylmorpholine

PyBOP benzotriazol-1-yl-oxytripyrrolidinyl hexafluorophosphate

THF tetrahydrofuran

NaN₃Sodium azide

MeOH methanol

TsCl-p-methylbenzenesulfonyl chloride

Pyr pyridine

CH₂Cl₂Methylene dichloride

Et₃N-Triethylamine

DMAP 4-dimethylaminopyridine

DCC dicyclohexylcarbodiimide

TsCl tosyl chloride

PPTS-pyridinium p-toluenesulfonate

AllocBt allyl benzotriazole carbonate.

Drawings

FIG. 1 is a synthetic scheme of

compounds

10a, 12a, 14 and 16 of the present invention.

FIG. 2 structural formulas of the compounds 1 to 8 of the present invention.

FIG. 3 is a scheme of synthesis of compounds 1-5 of the present invention.

FIG. 4 is a synthetic scheme of Compounds 6-8 of the present invention.

FIG. 5 Compounds 1-8 of the present invention or vitamin C on PC12 cells in H₂O₂Protective Effect under Damage, # P compared to control ####<0.001; comparing with model<0.001。

FIG. 6 protection of PC12 cells by compounds 1-8 of the invention in the case of 6-OHDA damage, ## P <0.001 compared to controls; compared to the model, P < 0.001.

FIG. 7Annexin V-PI double staining method for detecting neuroprotective effect of

compounds

5,6 and 8 of the present invention.

FIG. 8 Effect of

Compounds

5,6 and 8 of the present invention on intracellular ROS levels Experimental results.

Detailed Description

The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. These examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.

Example 1 Synthesis of glycosyl ligands (9a,9b) and cinnamic acid derivatives (10a,11a,11b,12a)

Reagents and conditions (a)9a allyl bromide, K₂CO₃Acetone, reflux overnight, 89%; 9b isopropylamine, NaOH, Ac₂O,H₂O,81 percent; (b) i) allyl bromide, K₂CO₃Acetone, refluxing overnight; ii) MeOH, saturated NaHCO₃Refluxing the solution for 2h, 88% for 10a and 80% for 11 a; 11b DMAP, pyridine, Ac₂O, 86%; (c) malonic acid, piperidine, pyridine, refluxed overnight, 70%.

Synthesis of 3, 4-Diallyloxyphenylethanol (9a)

To a 100mL round bottom flask was added 3, 4-dihydroxybenzene ethanol (9,0.83g,5.4mmol,1.0equiv), allyl bromide (2.59g,21.6mmol,4.0equiv), K₂CO₃(3.73g,27mmol,5.0equiv), acetone (50mL), heated at reflux for 12h, TLC to monitor completion of the reaction, filtered, concentrated, and column chromatographed (EtOAc/petroleum ether: 3/1) to give 1.12g of product as a colorless oil in 89% yield.

¹H NMR(400MHz,CDCl₃)6.84(d,J＝8.0Hz,1H),6.78-6.73(m,2H),6.12-6.02(m,2H),5.44-5.36(m,2H),5.28-5.24(m,2H),4.62-4.56(m,4H),3.81(t,J＝6.5Hz,2H),2.78(t,J＝6.5Hz,2H)；¹³C NMR(100MHz,CDCl₃)148.6,147.2,133.6,133.5131.5,121.4,117.5,117.4,115.3,114.6, 70.1,70.0,63.6, 38.6; MS (m/z) calculated value C₁₄H₁₈O₃:234；found:234[M]⁺(ii) a The elements comprise: calculated value C₁₄H₁₈O₃C in percentage, 71.77; h, 7.74; found C, 71.72; h,7.84.

Synthesis of 3, 4-diacetylphenethyl alcohol (9b)

0.5g of NaOH was put into a 100mL round-bottom flask, 1.5mL of water was added and dissolved, 30mL of isopropylamine, 3, 4-dihydroxybenzene ethanol (9, 1.0g,6.49mmol,1equiv), and 1.6mL of acetic anhydride were added all at once and stirred at room temperature for 40 min. Diluted with 150mL of ethyl acetate, extracted with ice water/ethyl acetate and neutralized with acetic acid. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (ethyl acetate/petroleum ether: 1/3) to obtain an oily substance 1.13g with a yield of 81%.¹H NMR(400MHz,CDCl₃)7.11(s,2H),7.05(s,1H),3.82(t,J＝6.3Hz,2H),2.83(t,J＝6.3Hz,2H),2.28(s,6H),1.76(br,1H)；¹³C NMR(100MHz,CDCl₃)168.42,168.36,141.9,140.5,137.7,127.2,123.8,123.3,63.2,38.5,20.6；ESI-HRMS[M+Na]⁺Calculated value C₁₂H₁₄NaO₅261.0733, found 261.0730.

Synthesis of 3, 4-diallyloxyphenylacetic acid (10a) and 3, 4-diallyloxycinnamic acid (11a)

In a 250ml round bottom flask, add acid compound 10 or compound 11(1.0equiv), allyl bromide (4.0equiv), anhydrous K₂CO₃(5.0equiv) and anhydrous acetone, and heating and refluxing for 12h with stirring. Insoluble matter was filtered off and concentrated. Saturated NaHCO was added directly to the residue₃The aqueous solution and methanol (1:1) were refluxed with stirring for 1 hour. The reaction solution was cooled to room temperature and the pH was adjusted to 6 with 1mol/L hydrochloric acid. Filtering under reduced pressure to obtain yellow solid. Further recrystallization from a mixed solvent of petroleum ether and ethyl acetate gave a white solid.

Compound 10a was prepared from compound 10 in 88% yield.¹H NMR(400MHz,CDCl₃)6.89-6.86(m,2H),6.83(dd,J＝1.9Hz,8.2Hz,1H),6.15-6.05(m,2H),5.47-5.41(m,2H),5.31-5.29(m,2H),4.63-4.62(m,4H),3.60(s,2H)；¹³C NMR(100MHz,CDCl₃)177.2,148.6,148.0,133.5,133.4,126.2,122.0,117.6,117.5,115.5,114.4,70.1,70.0, 40.5; MS (m/z) calculated value C₁₄H₁₆O₄248, found 248M]⁺Elemental composition: calculated value C₁₄H₁₆O₄(%) C, 67.73; h, 6.50; found C, 67.51; h,6.75.

Compound 11a was prepared from compound 11 in 80% yield.¹H NMR(400MHz,CDCl₃)7.70(d,J＝15.9Hz,1H),7.12-7.10(m,2H),6.88(d,J＝8.1Hz,1H),6.28(d,J＝15.9Hz,1H),6.08(m,2H),5.47-5.41(m,2H),5.33-5.29(m,2H),4.66-4.64(m,4H)；¹³C NMR(100MHz,CDCl₃)172.5,151.1,148.6,146.9,133.0,132.8,127.2,123.1,118.0,117.9,114.9,113.4,112.9,70.0,69.7；ESI-HRMS[M+H]⁺Calculated value C₁₅H₁₇O₄261.1121, found 261.1121.

Synthesis of 3, 4-diacetylcinnamic acid (11b)

Compound 11(1.0g,5.56mmol,1.0equiv) was dissolved in 50mL of pyridine solution, and Ac was added₂O (2mL), and stirred at room temperature for 3 hours. The solvent was evaporated to dryness and the residue was chromatographed on silica gel (ethyl acetate/petroleum ether: 1/1) to give 1.26 g of a white solid in 86% yield.¹H NMR(400MHz,CDCl₃)7.65(d,J＝15.9Hz,1H),7.36(d,J＝8.4Hz,1H),7.32(s,1H),7.18(d,J＝9.2Hz,1H),6.32(d,J＝15.9Hz,1H),2.24(s,3H),2.23(s,3H)；¹³CNMR(100MHz,CDCl₃)171.3,168.0,145.0,143.8,142.5,132.9,126.7,124.0,123.0,118.4,20.64,20.59；ESI-HRMS[M+K]⁺Calculated value C₁₃H₁₂KO₆303.0842, found 303.0839.

Compound 12a was prepared according to the method disclosed in the eur.j.pharm.sci.,23(2004)363-369.

EXAMPLE 2 Synthesis of (3, 4-Diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 16)

Reagents and conditions (a) TMSOTf,9a, CH₂Cl₂,

MS,-72℃,93％；(b)PPTS,MeCN,H₂O, reflux, 99%; (c) AllocBt, Et₃N,CH₂Cl₂,92％.

Synthesis of (3, 4-Diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-4, 6-O-benzylidene-beta-D-glucopyranoside (Compound 14)

Adding into a 250mL two-mouth bottle

MS (750mg), after baking and cooling, compound 13(0.50g,10.1mmol,1.0equiv) { reference J.org.chem.,65(2000) 2410-2431; J.am.chem.Soc.,121(1999) 734-753; tetrahedron Lett.,29(1988) 2299-prepar } and Compound 9a (0.25g,10.6mmol,1.05equiv), oil pump vacuum, Ar protection, 60mL of dried DCM, TMSOTf (20uL) at-40 deg.C, TLC after 2h to monitor completion of the reaction, low temperatureTriethylamine was extracted, filtered through celite, concentrated under reduced pressure, and column chromatographed (petroleum ether/ethyl acetate 3:1) to give 0.53g of an oily substance in 93% yield.

[α]_D ²⁵-17.1(c 0.0203,CDCl₃)；¹H NMR(400MHz,CDCl₃)7.44-7.41(m,2H),7.35-7.34(m,3H),6.81(d,J＝8.1Hz,1H),6.74(d,J＝1.6Hz,1H),6.71(dd,J＝1.6Hz,8.1Hz.1H),6.13-6.01(m,2H),5.48(s,1H),5.44-5.36(m,2H),5.31-5.23(m,3H),4.99(t,J＝9.1Hz,7.9Hz,1H),4.60-4.56(m,5H),4.35(dd,J＝4.9Hz,10.5Hz,1H),4.07(dt,J＝6.5Hz,9.4Hz,1H),3.77(t,J＝10.3Hz,1H),3.71-3.62(m,2H),3.51(dt,J＝4.8Hz,9.8Hz,1H),2.80(t,J＝6.4Hz,2H),2.03(s,3H),1.82(s,3H)；¹³C NMR(100MHz,CDCl₃)170.1,169.5,148.4,147.1,145.6,136.8,133.6,133.5,131.0,129.3,128.2,126.1,124.7,121.3,117.43,117.37,115.3,114.4,101.5,101.3,78.3,72.1,71.7,70.9,70.1,69.9,68.5,66.3,35.5,20.7,20.5；ESI-HRMS[M+H]⁺Calculated value C₃₁H₃₇O₁₀569.2381, found 569.2381.

(3, 4-Diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-. beta. -D-glucopyranoside (Compound 15)

A250 mL round bottom flask was charged with Compound 14(1.00g,1.76mmol,1.0equiv), acetonitrile 90mL and water 10mL, as well as PPTS (840mg,3.35mmol,2.0equiv), heated with stirring at reflux for 12 hours, and after completion of the reaction monitored by TLC, NaHCO₃Washing, H₂Washing with O and Na₂SO₄Drying, filtration, concentration and column chromatography of the residue (petroleum ether: ethyl acetate 1:1) gave 770mg of an oily substance in 99% yield. [ alpha ] to]_D ²⁵-12.8(c 0.027,CDCl₃)；¹H NMR(400MHz,CDCl₃)6.80(d,J＝8.1Hz,1H),6.73(d,J＝1.8Hz,1H),6.70(d,J＝1.8Hz,8.1Hz,1H),6.13-6.01(m,2H),5.44-5.36(m,2H),5.28-5.23(m,2H),4.99(dd,J＝9.5Hz,9.2Hz,1H),4.91(dd,J＝7.8Hz,9.7Hz,1H),4.61-4.56(m,4H),4.49(d,J＝7.8Hz,1H),4.06(dt,J＝6.5Hz,9.5Hz,1H),3.92(dd,J＝3.4Hz,11.9Hz,1H),3.82(dd,J＝4.9Hz,12.0Hz,1H),3.75(t,J＝9.2Hz,1H),3.64(dt,J＝7.3Hz,9.4Hz,1H),3.43-3.39(m,1H),2.79(t,J＝6.8Hz,2H),2.08(s,3H),1.91(s,3H)；¹³C NMR(100MHz,CDCl₃)171.8,169.5,148.5,147.2,133.7,133.6,131.5,121.4,117.5,117.4,115.5,114.5,100.8,76.2,75.5,71.1,70.8,70.2,70.0,69.6,62.2,35.6,20.8,20.6；ESI-HRMS[M+Na]⁺Calculated value C₂₄H₃₂NaO₁₀503.1888, found 503.1890.

(3, 4-Diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 16)

Compound 15(100mg,0.208mmol,1.0equiv), AocoBT (54mg,0.250mmol,1.2equiv), 10mL of dried DCM was charged into a 25mL flask, and Et was added with stirring₃N (140uL, 1mmol) TLC monitored for 8h reaction completion, neutralized with acetic acid and spin dried, and column chromatography (petroleum ether: ethyl acetate: 2:1) afforded compound 114mg yield 92%. [ alpha ] to]_D ²⁵-27.1(c0.008,CDCl₃)；¹H NMR(400MHz,CDCl₃)6.80(d,J＝8.1Hz,1H),6.73(d,J＝1.8Hz,1H),6.70(d,J＝1.8Hz,8.1Hz,1H),6.13-6.01(m,2H),5.97-5.87(m,1H),5.45-5.34(m,3H),5.29-5.23(m,3H),4.99(d,J＝8.7Hz,1H),4.92(d,J＝7.8Hz,9.6Hz,1H),4.64-4.62(m,2H),4.60-4.55(m,4H),4.47-4.44(m,3H),4.06(dt,J＝6.5Hz,9.5Hz,1H),3.68-3.58(m,2H),3.56-3.52(m,1H),2.90(d,J＝3.1Hz,1H),2.79(d,J＝6.9Hz,2H),2.08(s,3H),1.91(s,3H)；¹³C NMR(100MHz,CDCl₃)171.6,169.4,155.3,148.4,147.1,133.7,133.6,131.6,131.3,121.4,119.2,117.5,117.4,115.4,114.5,100.7,75.8,74.0,71.0,70.7,70.2,70.0,69.1,68.9,66.3,35.5,20.8,20.6；ESI-HRMS[M+Na]⁺Calculated value C₂₈H₃₆NaO₁₂587.2103, found 587.2104.

EXAMPLE 3 Synthesis of Compounds 1-5

Reagents and conditions (a) DCC, DMAP, CH₂Cl₂,0℃ to r.t.；(b)AcCl,MeOH,CH₂Cl₂；(c)10％Pd/C,MeOH,H₂O,HClO₄Or TsOH.

General synthetic method for compounds 18a-e

Compound 16(1.0equiv) and the corresponding acid (1.5equiv) were added dissolved in dichloromethane, DCC (1.5equiv), DMAP (1.5equiv) were added at 0 ℃ and then stirred at room temperature for 12 hours. The solvent was evaporated to dryness and the residue was chromatographed (petroleum ether: ethyl acetate 1:4 to 1:3) to give the corresponding product.

Synthesis of (3, 4-Diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-4-O- (3, 4-diallyloxyphenyl) levulinyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 18a)

Following the general procedure, compounds 16 and 9a were coupled to give compound 18a as a colorless oil in 85% yield. [ alpha ] to]_D ²⁵-9.8(c 0.008,CDCl₃)；¹H NMR(400MHz,CDCl₃)7.59(d,J＝15.9Hz,1H),7.08-7.05(m,2H),6.87(d,J＝8.1Hz,1H),6.80(d,J＝8.1Hz,1H),6.75(d,J＝1.8Hz,1H),6.71(dd,J＝1.8Hz,8.1Hz,1H),6.18(d,J＝15.9Hz,1H),6.13-6.01(m,4H),5.94-5.84(m,1H),5.46-5.21(m,11H),5.15(t,J＝9.6Hz,1H),5.01(dd,J＝8.0Hz,9.6Hz,1H),4.65-4.56(m,10H),4.52(d,J＝7.9Hz,1H),4.22-4.26(m,2H),4.08(dt,J＝6.4Hz,9,4Hz,1H),3.81-3.77(m,1H),3.65(dt,J＝6.4Hz,9,4Hz,1H),2.80(t,J＝6.8Hz,2H),1.96(s,3H),1.91(s,3H)；¹³CNMR(100MHz,CDCl₃)170.2,169.3,165.6,154.6,151.1,148.7,148.5,147.2,146.6,133.7,133.6,133.0,132.8,131.6,131.4,127.1,123.2,121.4,119.0,118.0,117.9,117.4,117.3,115.5,114.6,113.9,113.5,112.9,100.7,72.6,72.1,71.3,70.7,70.2,70.03,70.00,69.7,68.7,66.2,35.6,20.6,20.5；ESI-HRMS[M+Na]⁺Calculated value C₄₃H₅₀NaO₁₅829.3042, found 829.3052.

Synthesis of (3, 4-Diallyloxyphenylethyl) 2, 3-O-acetyl-4-O- (3, 4-difluorocinnamoyl) -6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 18b)

Following the general procedure, compounds 16 and 17a (available from carbofuran) were coupled to give compound 18b as a colorless oil in 86% yield. [ alpha ] to]_D ²⁵-16.5(c 0.018,CDCl₃)；¹H NMR(400MHz,CDCl₃)7.58(d,J＝15.9Hz,1H),7.36-7.31(m,1H),7.25-7.15(m,2H),6.81(d,J＝8.2Hz,1H),6.75(d,J＝1.9Hz,1H),6.71(dd,J＝1.9Hz,8.3Hz,1H),6.27(d,J＝15.9Hz,1H),6.13-6.01(m,2H),5.94-5.84(m,1H),5.45-5.22(m,7H),5.15(t,J＝9.6Hz,1H),5.02(dd,J＝8.0Hz,9.6Hz,1H),4.64-4.55(m,6H),4.53(d,J＝7.9Hz,1H),4.31-4.24(m,2H),4.09(dt,J＝6.4Hz,9.5Hz,1H),3.82-3.77(m,1H),3.65(dt,J＝6.4Hz,9.5Hz,1H),2.81(t,J＝6.8Hz,2H),1.97(s,3H),1.92(s,3H)；¹³C NMR(100MHz,CDCl₃)170.2,169.2,164.8,154.6,148.5,147.1,144.2,133.7,133.6,131.6,131.3,125.1,121.4,119.0,118.0,117.8,117.5,117.4,116.7,116.5,115.4,114.6,100.7,72.5,71.9,71.2,70.8,70.2,70.0,69.1,68.7,66.0,35.5,20.55,20.48；ESI-HRMS[M+Na]⁺Calculated value C₃₇H₄₀NaO₁₃F₂753.2329, found 753.2337.

Synthesis of (3, 4-Diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-4-O- (3, 4-dimethoxycinnamoyl) -6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 18c)

Following the general procedure, compounds 16 and 17b (available from carbofuran) were coupled to give compound 18c as a colorless oil in 98% yield. [ alpha ] to]_D ²⁵-9.3(c 0.013,CDCl₃)；¹H NMR(400MHz,CDCl₃)7.63(d,J＝15.9Hz,1H),7.11-7.03(m,2H),6.86(d,J＝8.3Hz,1H),6.81(d,J＝8.2Hz,1H),6.74-6.70(m,2H),6.22(d,J＝15.9Hz,1H),6.13-6.02(m,2H),5.94-5.85(m,1H),5.45-5.21(m,7H),5.15(t,J＝9.6Hz,1H),5.02(dd,J＝8.1Hz,9.5Hz,1H),4.60-4.52(m,7H),4.28-4.27(m,2H),4.09(dt,J＝6.3Hz,9.4Hz,1H),3.91(s,6H),3.82-3.77(m,1H),3.65(dt,J＝7.4Hz,9.4Hz,1H),2.81(t,J＝6.8Hz,2H),1.97(s,3H),1.89(s,3H)；¹³C NMR(100MHz,CDCl₃)170.2,169.2,165.6,154.6,151.6,149.3,148.5,147.1,146.7,133.7,133.6,131.6,131.3,126.9,123.2,121.4,119.0,117.4,117.3,115.4,114.6,113.8,111.0,109.7,100.7,72.7,72.6,72.1,71.3,70.7,70.2,69.9,68.7,66.2,56.0,55.9,35.5,20.6 20.5；ESI-HRMS[M+Na]⁺Calculated value C₃₉H₄₆NaO₁₅777.2729, found 777.2764.

Synthesis of (3, 4-Diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-4-O-p-chlorocinnamoyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 18D)

Following the general procedure, compounds 16 and 12a were coupled to give compound 18d as a colorless oil in 87% yield. [ alpha ] to]_D ²⁵-22.6(c 0.019,CDCl₃)；¹H NMR(400MHz,CDCl₃)7.62(d,J＝15.9Hz,1H),7.44(d,J＝8.5Hz,2H),7.36(d,J＝8.5Hz,2H),6.81(d,J＝8.4Hz,1H),6.74(d,J＝1.9Hz,1H),6.71(d,J＝1.9Hz,8.2Hz,1H),6.33(d,J＝15.9Hz,1H),6.12-6.02(m,2H),5.94-5.84(m,1H),5.45-5.36(m,2H),5.34-5.21(m,5H),5.15(t,J＝9.6Hz,1H),5.01(dd,J＝8.0Hz,9.6Hz,1H),4.62-4.55(m,6H),4.53(d,J＝7.9Hz,1H),4.27-4.25(m,2H),4.08(dt,J＝6.4Hz,9.5Hz,1H),3.80(m,1H),3.65(ddd,J＝7.3Hz,7.5Hz,9.5Hz,1H),2.80(d,J＝6.8Hz,2H),1.96(s,3H),1.91(s,3H)；¹³C NMR(100MHz,CDCl₃)170.2,169.3,165.1,154.6,147.1,145.2,136.8,133.7,133.6,132.4,131.5,131.3,129.5,129.2,121.4,119.0,117.5,117.45,117.4,116.8,115.3,114.5,100.7,72.5,71.9,71.2,70.8,70.2,69.9,69.0,68.8,66.1,35.5,20.6,20.5；ESI-HRMS[M+NH₄]⁺Calculated value C₃₇H₄₅NO₁₃Cl746.2574, found 746.2564.

Synthesis of (3, 4-Diallyloxyphenyl) ethyl 2, 3-di-O-acetyl-4-O- (3, 4-allyloxyphenyl) acetyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 18e)

Following the general procedure, coupling of

compounds

16 and 10a gave compound 18e as a colorless oil in 95% yield. [ alpha ] to]_D ²⁵-10.3(c 0.016,CDCl₃)；¹H NMR(400MHz,CDCl₃)6.85-6.68(m,6H),6.12-6.00(m,4H),5.95-5.86(m,1H),5.46-5.32(m,5H),5.29-5.22(m,5H),5.18(t,J＝9.5Hz,1H),5.00(dd,J＝9.6Hz,9.8Hz,1H),4.93(dd,J＝9.0Hz,9.7Hz,1H),4.62-4.55(m,10H),4.47(d,J＝8.0Hz,1H),4.21-4.10(m,2H),4.05(dt,J＝6.5Hz,9.5Hz,1H),3.71-3.66(m,1H),3.62(dt,J＝9.5Hz,6.5Hz,1H),3.48(dd,J＝14.6Hz,2.6Hz,2H),2.78(t,J＝6.8Hz,2H),1.88(s,3H),1.75(s,3H)；¹³C NMR(100MHz,CDCl₃)170.3,170.0,169.1,154.5,148.6,148.4,147.9,147.1.133.7,133.6,133.4,133.3,131.5,131.3,126.0,121.8,119.0,117.5,117.46,117.37,117.3,115.4,115.2,114.6,114.4,100.6,72.3,71.7,71.2,70.7,70.1,70.0,69.93,69.87,68.9,68.7,65.8,40.5,35.5,20.4,20.3；ESI-HRMS[M+NH₄]⁺Calculated value C₄₂H₅₄NO₁₅812.3488, found 812.3465.

General synthetic method for compounds 19a-e

Compound 18a-e was dissolved in a mixed solvent (dichloromethane: methanol ═ 1:1), acetyl chloride was added, and the mixture was stirred at room temperature for 12 hours. TLC to monitor reaction completion, add a small amount of saturated NaHCO₃Neutralizing the solution, concentrating, and separating the residue by column chromatography to obtain the corresponding product.

Synthesis of 3, 4-Diallyloxyphenyl) ethyl 4-O- (3, 4-diallyloxycinnamoyl) -6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 19a)

Compound 18a was deacetylated according to the general procedure to give (eluent petroleum ether: ethyl acetate ═ 2:1) 19d as a white solid in 91% yield. [ alpha ] to]_D ²⁵-3.2(c 0.018,CDCl₃)；¹H NMR(400MHz,CDCl₃)7.67-7.63(d,J＝15.9Hz,1H),7.07-7.06(m,2H),6.87-6.73(m,4H),6.26(d,J＝15.9Hz,1H),6.12-6.01(m,4H),5.91-5.83(m,1H),5.44-5.20(m,10H),4.96(t,J＝9.6Hz,1H),4.64-4.56(m,10H),4.33-4.27(m,3H),4.10(dt,J＝6.9Hz,9.3Hz,1H),3.76-3.69(m,3H),3.49(dd,J＝8.3Hz,8.7Hz,1H),2.94(br,1H),2.87(t,J＝7.5Hz,2H),2.60(br,1H)；¹³C NMR(125MHz,CDCl₃):166.8,154.7,151.0,148.51,148.48,147.1,146.6,133.6,133.5,133.0,132.8,131.3,131.2,127.1,123.1,121.3,119.1,118.0,117.9,117.5,117.4,115.1,114.4,114.2,113.3,112.7,102.4,74.6,74.0,72.1,70.9,70.1,70.0,69.9,69.7,68.7,66.5,35.6；ESI-HRMS[M+H]⁺Calculated value C₃₉H₄₇O₁₃723.3011, found 723.2986.

Synthesis of (3, 4-Diallyloxyphenylethyl) 4-O- (3, 4-difluorocinnamoyl) -6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 19b)

Deacetylation at 18b according to general procedure gave (eluent petroleum ether: ethyl acetate: 3:1) 19b as a white solid in 98% yield. [ alpha ] to]_D ²⁵-4.8(c 0.002,CDCl₃)；¹H NMR(400MHz,CDCl₃)7.63(d,J＝15.9Hz,1H),7.37-7.32(m,1H),7.28-7.24(m,1H),7.24-7.15(m,1H),6.82(d,J＝8.1Hz,1H),6.78-6.73(m,2H),6.36(d,J＝15.9Hz,1H),6.13-6.02(m,2H),5.93-5.83(m,1H),5.44-5.21(m,6H),4.99(t,J＝9.7Hz,1H),4.60-4.57(m,6H),4.32(d,J＝7.8Hz,1H),4.28-4.26(m,2H),4.12(dt,J＝7.1Hz,9.6Hz,1H),3.77-3.68(m,3H),3.48(dt,J＝2.2Hz,8.5Hz,1H),2.87(t,J＝7.6Hz,2H),2.76(d,J＝3.5Hz,1H),2.49(d,J＝2.2Hz,1H)；¹³C NMR(100MHz,CDCl₃)165.9,154.7,148.6,147.3,144.2,133.6,133.58,131.4,131.3,125.0,121.4,119.0,118.0,117.95,117.94,117.8,117.5,117.4,116.6,116.4,115.3,114.6,102.5,74.5,74.1,72.1,71.2,71.0,70.2,70.0,68.7,66.4,35.6；ESI-HRMS[M+NH₄]⁺Calculated value C₃₃H₄₀NO₁₁F₂664.2564, found 664.2565.

Synthesis of (3, 4-Diallyloxyphenylethyl) 4-O- (3, 4-dimethoxycinnamoyl) -6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 19c)

Deacetylation via 18c according to the general procedure gave (eluent petroleum ether: ethyl acetate ═ 2:1) 19c as a white solid in 90% yield. [ alpha ] to]_D ²⁵-48.0(c 0.0007,CDCl₃)；¹H NMR(400MHz,CDCl₃)7.68(d,J＝15.9Hz,1H),7.11-7.05(m,2H),6.87-6.73(m,4H),6.31(d,J＝15.9Hz,1H),6.13-6.02(m,2H),5.93-5.83(m,1H),5.44-5.21(m,6H),4.97(t,J＝9.6Hz,1H),4.60-4.57(m,5H),4.33(d,J＝7.8Hz,1H),4.29(m,2H),4.11(dt,J＝7.0Hz,9.4Hz,1H),3.92-3.88(m,6H),3.80-3.69(m,4H),3.50(t,J＝9.2Hz,1H),2.97(d,J＝3.0Hz,1H),2.87(t,J＝7.6Hz,2H),2.60(s,1H)；¹³C NMR(100MHz,CDCl₃)166.8,154.7,151.5,149.3,148.6,147.2,146.7,133.63,133.57,131.4,131.3,127.0,123.1,121.4,119.0,117.5,117.4,115.3,114.5,114.2,111.1,109.8,102.5,74.7,74.1,72.1,71.02,70.96,70.1,70.0,66.5,56.0,55.9,35.6；ESI-HRMS[M+H]⁺Calculated value C₃₅H₄₃O₁₃671.2698, found 671.2698.

Synthesis of (3, 4-Diallyloxyphenyl) ethyl 4-O-p-chlorocinnamoyl-6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 19D)

Deacetylation over 18d according to general procedure gave (eluent petroleum ether: ethyl acetate ═ 2:1) 19d as a white solid in 98% yield. [ alpha ] to]_D ²⁵-12.5(c 0.004,CDCl₃)；¹H NMR(400MHz,CDCl₃)7.69(d,J＝16.0Hz,1H),7.45(m,2H),7.37(m,2H),6.82(d,J＝8.1Hz,1H),6.78(d,J＝1.6Hz,1H),6.74(dd,J＝1.6Hz,8.0Hz,1H),6.41(d,J＝16.0Hz,1H),6.13-6.02(m,2H),5.93-5.83(m,1H),5.45-5.37(m,2H),5.35-5.20(m,4H),4.99(t,J＝9.6Hz,1H),4.61-4.56(m,6H),4.32(d,J＝7.7Hz,1H),4.28-4.27(m,2H),4.12(dt,J＝7.2Hz,9.6Hz,1H),3.77-3.68(m,3H),3.51-3.46(m,1H),2.87(t,J＝7.8Hz,2H),2.74(d,J＝3.3Hz,1H),2.43(d,J＝2.0Hz,1H)；¹³C NMR(100MHz,CDCl₃)166.2,154.7,148.6,147.3,145.2,136.7,133.7,133.6,132.5,131.4,131.3,129.3,121.4,119.0,117.5,117.4,117.3,115.3,114.5,102.5,74.5,74.1,72.1,71.1,71.0,70.2,70.0,68.7,66.4,35.6；ESI-HRMS[M+H]⁺Calculated value C₃₃H₃₈O₁₁Cl 645.2097, found 645.2071.

Synthesis of (3, 4-Diallyloxyphenyl) ethyl 4-O- (3, 4-diallyloxyphenylacetyl) -6-O-allyloxycarbonyl-beta-D-glucopyranoside (Compound 19e)

Deacetylation of 18e according to the general procedure gave (eluent petroleum ether: ethyl acetate ═ 2:1) 19e as a white solid in 99% yield. [ alpha ] to]_D ²⁵-7.6(c 0.003,CDCl₃)；¹H NMR(400MHz,CDCl₃)6.87-6.71(m,6H),6.12-6.01(m,4H),5.94-5.84(m,1H),5.45-5.35(m,5H),5.30-5.23(m,5H),4.84(t,J＝9.6Hz,1H),4.57(br,10H),4.25(d,J＝7.7Hz,1H),4.17(dd,J＝5.9Hz,11.8Hz,1H),4.10-4.04(m,2H),3.71-3.57(m,5H),3.41(dd,J＝8.1Hz,8.8Hz,1H),2.84(t,J＝7.4Hz,2H)；¹³CNMR(125MHz,CDCl₃)171.4,154.6,148.6,148.5,147.8,147.2,133.6,133.5,133.44,133.36,131.2,126.2,121.3,119.1,117.6,117.54,117.49,115.2,115.1,114.4,114.3,102.4,74.4,74.0,71.9,71.2,71.0,70.1,71.0,70.1,70.0,69.9,68.7,66.1,40.7,35.6；ESI-HRMS[M+NH₄]⁺Calculated value C₃₈H₅₀NO₁₃728.3277, found 728.3290.

General synthetic methods for compounds 1-5:

compounds 19a-e were dissolved in a mixed solvent (dichloromethane: methanol 20:1) and 10% was addedPd/C and catalytic amount of p-toluenesulfonic acid (or HClO)₄) And reacting at 60 ℃ for 12 hours. Filtration, concentration and preparative thin-layer separation (dichloromethane/methanol, 10:1 to 5:1) gave the corresponding product.

Synthesis of (3, 4-dihydroxyphenyl) ethyl 4-O- (3, 4-dihydroxycinnamoyl) -beta-D-glucopyranoside (Compound 1)

Compound 19a was reacted according to the general procedure and isolated (dichloromethane: methanol: 8:1) to give compound 1 as a yellow oil in 62% yield. [ alpha ] to]_D ²⁵-25.7(c 0.026,MeOH)；¹H NMR(400MHz,CD₃OD)7.58(d,J＝15.8Hz,1H),7.04(d,J＝1.3Hz,1H),6.94(d,J＝8.1Hz,1H),6.79(d,J＝8.2Hz,1H),6.70(m,2H),6.58(m,1H),6.28(d,J＝15.8Hz,1H),4.35(d,J＝7.8Hz,1H),4.08(m,2H),3.74(m,1H),3.65(m,2H),3.55(m,2H),3.35-3.25(m,1H),2.81(t,J＝6.5Hz,2H)；¹³C NMR(100MHz,CD₃OD)168.6,149.7,147.6,146.8,146.1,144.6,131.5,127.7,123.1,121.3,117.1,116.5,116.3,115.2,114.7,104.4,76.1,75.8,75.2,72.5,72.2,62.5,36.6；ESI-HRMS[M+Na]⁺Calculated value C₂₃H₂₆NaO₁₁501.1367, found 501.1369.

Synthesis of (3, 4-diallyloxyphenethyl) 4-O- (3, 4-difluorocinnamoyl) -beta-D-glucopyranoside (Compound 2)

Compound 19b was reacted according to the general procedure and isolated (dichloromethane: methanol: 10:1) to give compound 2 as a colorless oil in 35% yield. [ alpha ] to]_D ²⁵-10.6(c 0.012,CD₃OD)；¹H NMR(400MHz,CD₃OD)7.68(d,J＝16.0Hz,1H),7.66-7.58(m,1H),7.46-7.43(m,1H),7.34-7.27(m,1H),6.70-6.66(m,2H),6.58-6.54(m,2H),4.88(t,J＝9.4Hz,1H),4.36(d,J＝7.8Hz,1H),4.08-4.02(m,1H),3.75-3.70(m,1H),3.64(m,2H),3.58-3.49(m,2H),3.35-3.28(m,1H),2.80(dt,J＝2.3Hz,7.5Hz,2H)；¹³C NMR(100MHz,CD₃OD)167.5,146.1,144.7,144.4,133.4,131.5,126.6,126.5,121.3,120.1,119.0,118.8,117.7,117.5,117.1,116.3,104.4,76.0,75.8,75.2,72.9,72.2,62.4,36.6；ESI-HRMS[M+Na]⁺Calculated value C₂₃H₂₄NaO₉F₂505.1281, found 505.1286.

Synthesis of (3, 4-hydroxyphenylethyl) 4-O- (3, 4-dimethoxycinnamoyl) -beta-D-glucopyranoside (Compound 3)

Compound 19c was reacted according to the general procedure and isolated (dichloromethane: methanol: 10:1) to give compound 3 as a yellow oil in 34% yield. [ alpha ] to]_D ²⁵-5.4(c 0.010,MeOH)；¹H NMR(400MHz,CD₃OD)7.67(d,J＝15.9Hz,1H),7.22(d,J＝1.9Hz,1H),7.17(dd,J＝1.8Hz,8.4Hz,1H),6.97(d,J＝8.4Hz,1H),6.68-6.65(m,2H),6.55(dd,J＝2.1Hz,8.1Hz,1H),6.44(d,J＝15.9Hz,1H),4.86(t,J＝9.4Hz,1H),4.36(d,J＝7.8Hz,1H),4.07-4.01(m,1H),3.85(m,6H),3.74-3.68(m,1H),3.64-3.58(m,2H),3.56-3.48(m,2H),3.32-3.28(m,1H),2.79(m,2H)；¹³C NMR(100MHz,CD₃OD)168.4,153.0,150.8,147.0,146.1,144.7,131.5,128.8,124.1,121.3,117.1,116.3,116.2,112.7,111.7,104.4,76.1,75.9,75.3,72.6,72.2,62.5,56.5,36.6；ESI-HRMS[M+H]⁺Calculated value C₂₅H₃₁O₁₁507.1861, found 507.1867 (3, 4-dihydroxyphenyl) ethyl 4-O-p-chlorocinnamoyl-beta-D-glucopyranoside Synthesis of (Compound 4)

Compound 19d was reacted according to the general procedure and isolated (dichloromethane: methanol: 12:1) to give compound 4 as a yellow oil in 32% yield. [ alpha ] to]_D ²⁵-13.7(c 0.010,MeOH)；¹H NMR(400MHz,CD₃OD)7.69(d,J＝16.0Hz,1H),7.59(d,J＝8.5Hz,2H),7.40(d,J＝8.5Hz,2H),6.69-6.65(m,2H),6.58-654(m,2H),4.87(t,J＝9.4Hz,1H),4.36(d,J＝7.8Hz,1H),4.08-4.01(m,1H),3.74-3.68(m,1H),3.66-3.61(m,2H),3.56-3.49(m,2H),3.33-3.28(m,1H),2.79(ddd,J＝2.3Hz,2.5Hz,7.7Hz,2H)；¹³C NMR(100MHz,CD₃OD)167.7,146.1,145.3,144.7,137.4,134.5,131.5,130.8,130.2,121.3,119.4,117.1 116.3,104.4,76.0,75.8,75.2,72.8,72.2,62.4,36.6；ESI-HRMS[M+NH₄]⁺Calculated value C₂₃H₂₉ClNO₉498.1525, found 498.1525.

Synthesis of (3, 4-dihydroxyphenylethyl) 4-O- (3, 4-dihydroxyphenylacetyl) -beta-D-glucopyranoside (Compound 5)

According to the general formulaAfter the reaction, compound 19e was isolated (dichloromethane: methanol: 8:1) to give compound 5 as a yellow oil in 37% yield. [ alpha ] to]_D ²⁵-6.4(c 0.004,MeOH)；¹H NMR(400MHz,CD₃OD)6.76-6.81(m,5H),6.56-6.62(m,1H),4.30(d,J＝7.6Hz,1H),4.14-3.98(m,2H),3.89-3.79(m,2H),3.65-3.76(m,3H),3.27-3.38(m,1H),3.15-3.22(m,2H),2.84(t,J＝6.8Hz,2H)；¹³C NMR(100MHz,CD₃OD)173.9,146.3,146.1,145.5,144.7,131.6,126.9,131.7,121.3,117.4,117.1,116.4,116.3,104.4,77.9,75.4,75.0,72.1,71.7,65.0,41.4,36.6；ESI-HRMS[M+Na]⁺Calculated value C₂₂H₂₆NaO₁₁489.1367, found 489.1376.

EXAMPLE 4 Synthesis of Compounds 6-8

Reagents and conditions (a) oxalyl chloride, CH₂Cl₂,Et₃N,69％；(b)AcCl,CH₂Cl₂,MeOH,82％；(c)i)10％Pd/C,MeOH,H₂O,HClO₄；ii)AcCl,CH₂Cl₂MeOH, overall yield of two steps 92%; (d) PyBOP, NMM, DMF, 39%.

Synthesis of 9' -decarboxylated tetraacetyl rosmarinic acid (Compound 20)

Compound 11b (100mg,0.379mmol,1equiv), 0.11mL oxalyl chloride was added to 2mL DCM, stirred at RT for 6h, and concentrated. Under Ar protection, compound 9b (90.2mg,0.379mmol,1.0equiv), DCM (5mL), Et was added₃N (0.5mL) was stirred at room temperature for 12 hours, and then the solvent was evaporated. Column chromatography of the residue (3: 1 petroleum ether/ethyl acetate) gave 126mg of oil in 69% yield.¹H NMR(400MHz,CDCl₃)7.60(d,J＝16.0Hz,1H),7.42-7.36(m,2H),7.22(d,J＝8.0Hz,1H),7.14-7.01(m,3H),6.36(d,J＝16.0Hz,1H),4.42(t,J＝6.7Hz,2H),3.00(t,J＝6.7Hz,2H),2.30(m,12H)；¹³C NMR(100MHz,CDCl₃)168.3,168.2,168.0,167.9,166.4,143.5,143.1,142.4,142.0,140.7,136.7,133.3,127.0,126.4,123.9,123.4,122.8,119.1,64.5,34.5,20.6,20.5；ESI-HRMS[M+Na]⁺Calculated value C₂₅H₂₄NaO₁₀507.1272, found 507.1262.

Synthesis of 9' -decarboxylated rosmarinic acid (Compound 6)

The compound 20 was used as a starting material to remove the acetyl protection (the same procedure as in the deacetylation step 1-5 of example 3). Column chromatography eluent dichloromethane: methanol 10:1 gave the compound as a yellow oil in 82% yield.¹H NMR(400MHz,CD₃OD)7.52(d,J＝15.8Hz,1H),7.04(s,1H),6.94(d,J＝8.1Hz,1H),6.78(d,J＝8.0Hz,1H),6.71(m,2H),6.58(d,J＝8.0Hz,1H),6.24(d,J＝15.8Hz,1H),4.30(t,J＝6.9Hz,2H),2.84(t,J＝6.9Hz,2H)；¹³C NMR(100MHz,CD₃OD)169.3,149.6,146.9,146.3,144.9,146.8,130.8,127.7,122.9,121.2,117.1,116.5,116.4,115.2,115.1,66.5,35.6；ESI-HRMS[M+Na]⁺Calculated value C₁₇H₁₆O₆Na 339.0842, found 339.0839.

Synthesis of (3, 4-dihydroxyphenylethyl) beta-D-glucopyranoside (Compound 7)

Compound 15(150mg,0.315mmol,1equiv) was dissolved in MeOH/H₂O (20:1,8mL), a catalytic amount of 10% Pd/C and a catalytic amount of HClO₄. The solvent was evaporated to dryness and the residue was deacetylated (same procedure as the deacetylation procedure of 1-5 in example 3) and then separated (eluent dichloromethane: methanol ═ 1:1) to give 12mg of a white solid in 92% yield. [ alpha ] to]_D ²⁵-0.59(c 0.025,CD₃OD)；¹H NMR(400MHz,CD₃OD)6.68-6.65(m,2H),6.55(d,J＝8.1Hz,1H),4.28(d,J＝7.3Hz,1H),4.01(dd,J＝8.6Hz,8.3Hz,1H),3.83(m,2H),3.75-3.70(m,1H),3.70-3.60(m,3H),3.17(t,J＝8.2Hz,1H),2.77(t,J＝7.3Hz,2H)；¹³C NMR(100MHz,CD₃OD)146.0,144.6,131.6,121.3,117.1,116.3,104,3,78.1,77.9,75.1,72.0,71.6,62.7,36.6；ESI-HRMS[M+H]⁺Calculated value C₁₄H₂₁O₈317.1231, found 317.1229.

Synthesis of 2-O-methyl-5-O- (p-hydroxycinnamoyl) -beta-D-sialoside (Compound 8)

Compound 21(100.0mg,0.34mmol,1.0equiv, synthesized as disclosed in Tetrahedron Asymmetry22(2011) 338-344) and compound 11(91.6mg,0.51mmol,1.5equiv) were dissolved in DMF (50mL), PyBOP (265.0mg,0.51mmol,1.5equiv) and NMM (103.0mg,1.02mmol,3.0equiv) were added and stirred at room temperature for 24 hoursThen, the mixture was concentrated, and the residue was separated by column chromatography (ethyl acetate: ethanol 10:1) to give 60mg of a yellow oil, yield 39%. [ alpha ] to]_D ²⁵-11.3(c 0.006,MeOH)；¹H NMR(400MHz,CD₃OD)7.44(d,J＝15.6Hz,1H),7.03(d,J＝1.7Hz,1H),6.91(dd,J＝2.1Hz,8.2Hz,1H),6.77(d,J＝8.2Hz,1H),6.43(d,J＝15.6Hz,1H),4.09(td,J＝4.8Hz,10.7Hz,1H),3.99(t,J＝10.2Hz,1H),3.81-3.91(m,3H),3.75-3.80(m,3H),3.68(dd,J＝5.0Hz,11.1Hz,1H),3.55(d,J＝8.0Hz,1H),3.29(s,3H),2.38(dd,J＝5.0Hz,12.9Hz,1H),1.69(dd,J＝1.6Hz,12.8Hz,1H)；¹³C NMR(100MHz,CD₃OD)171.1,170.6,148.9,146.8,143.0,128.2,122.3,118.0,116.5,115.0,100.5,72.6,71.4,70.1,67.7,65.2,53.8,53.3,51.7,41.6；ESI-HRMS[M-H]^-Calculated value C₂₀H₂₆NO₁₁456.1506, found 456.1511.

Experimental example 1 evaluation of cell protective Activity of the Compound of the present invention

1. Test compounds: compounds 1-8 prepared in example 3 and example 4.

2. Experimental Material

Rat adrenal pheochromocytoma cell line (PC12 cell) purchased from Shanghai cell bank of Chinese academy of sciences; 6-OHDA et al (sigma corporation); ROS kit (bi yun day biotechnology research institute); annexin V-FITC apoptosis detection kit (Kaikyi organism).

3. Experimental methods

Determination of cell viability: taking PC12 cells in logarithmic growth phase at 1 × 10⁵The density of (A) was laid in a 96-well plate with 100. mu.l per well, 3 parallel wells per group of samples, 5% CO ₂37 ℃ CO₂After 24 hours incubation in an incubator, test compounds were added for 6 hours pretreatment and 300. mu. M H added₂O₂After incubation at 37 ℃ for 1 hour or for 24 hours after 6-OHDA damage, 20. mu.l of MTT (5mg/ml) was added and incubation was continued for 4 hours, then DMSO solution was added and after the crystals were completely dissolved, the absorbance value was measured at a wavelength of 570nm using a microplate reader.

4. Results of the experiment

The results of the experiment are shown in FIGS. 5 and 6. Is about H₂O₂(FIG. 5) and 6-OHDA (FIG. 6) injury of PC12 cell model from the perspective of relative cell survival, the relative cell survival in the model group was significantly decreased compared to the normal control group, and there was a statistical difference (P)<0.05), indicating that the molding is successful. Compared with a model group, the compound 5, the compound 6 and the compound 8 have stronger cytoprotective effect and show obvious dose dependence, which indicates that the compound 5, the compound 6 and the compound 8 have better neuroprotective effect.

Experimental example 2 Annexin V-PI double staining method for testing neuroprotective effect of

compounds

5,6 and 8

1. Test compounds: (3, 4-dihydroxyphenylethyl) 4-O- (3, 4-hydroxyphenylethyl) - β -D-glucopyranoside (compound 5), 9' -decarboxylated rosmarinic acid (compound 6) and 2-O-methyl-5-O- (p-hydroxycinnamoyl) - β -D-sialoside (compound 8);

2. experimental methods

(1) Taking PC12 cells in logarithmic growth phase at 1 × 10⁵Was plated into 24-well plates containing coverslips at a density of 500. mu.l/well, 5% CO₂And incubated at 37 ℃ for 24 hours. After 6 hours of pretreatment with compound 5, compound 8, compound 6, cells were treated with 6-OHDA at a final concentration of 100. mu.M and cultured for 24 hours;

(2) cells were washed 2 times with PBS;

(3) adding 1 mul Annexin V-FITC and 1 mul PI into 100 mul Binding Buffer, and mixing uniformly;

(4) removing the culture medium, and dripping the solution on the surface of the cover glass to uniformly cover the surface of the cover glass;

(5) reacting for 5min at room temperature in the dark;

(6) the coverslip was inverted onto the slide and observed under confocal microscope: the Annexin V-FITC fluorescent signal is green, and the PI fluorescent signal is red.

3. Results of the experiment

From the results of detecting the effect of compound 5, compound 6 and compound 8 on apoptosis by Annexin V-PI double staining method (FIG. 7), normal control group Annexin V and PI are low stained, which indicates that the cells are in normal state; the model group Annexin V and PI are both high-stained, which indicates that PC12 cells enter the late apoptosis stage and necrotic cells appear after 6-OHDA injury is given; after the

compounds

5,6 and 8 are administered for pre-protection, the cells are controlled in the early apoptosis stage, and the cell damage condition is obviously reduced or even eliminated along with the increase of the administration dosage, which shows that the compound 5, the compound 6 and the compound 8 of the invention play a role in neuroprotection through inhibiting the apoptosis.

Experimental example 3 Effect of

Compounds

5,6 and 8 of the present invention on intracellular ROS levels

2. experimental methods

Taking PC12 cells in logarithmic growth phase at 1 × 10⁵The density of (A) was spread in a 96-well plate, 100. mu.l per well, 5% CO₂After 24 hours of incubation at 37 ℃, compounds 5,8 and 6 were added for pretreatment for 6 hours, followed by 6-OHDA addition for 24 hours of injury, the medium was removed, 40 μ l of DCFH-DA solution (stock solution diluted with serum-free medium at 1: 1000) was added to each well, the cell culture chamber was incubated for 30min in the dark at 37 ℃, washed three times with PBS, and observed under a fluorescent microscope.

3. Results of the experiment

As can be seen by measuring intracellular ROS levels (fig. 8), ROS are expressed in normal cells, but at lower levels; after 6-OHDA injury, intracellular ROS expression level is remarkably increased, which indicates that 6-OHDA causes intracellular oxidative stress; in contrast, in the cells to which compounds 5,6 and 8 of the present invention were administered, the expression level of ROS was significantly reduced compared to the model group, and the expression-inhibiting effect was positively correlated with the dose of the compound, indicating that

compounds

5,6 and 8 of the present invention exert neuroprotective effects by way of anti-oxidative stress.

In conclusion, the compound 5, the compound 6 and the compound 8 can obviously reduce the damage of 6-OHDA to cells in a dose-dependent manner through anti-apoptosis, anti-oxidative damage and other ways, play a role in neuroprotection and have no obvious toxic or side effect. Thus, the compound 5, the compound 6 and the compound 8 have great development value as potential new drugs for preventing and treating the neurological diseases.

Claims

1. A sialic acid methyl ester methyl glycoside derivative, which is a compound represented by the formula (III):

wherein R is₁Is 3, 4-dihydroxy cinnamoyl.

2. A process for the synthesis of a compound of formula iii according to claim 1, comprising the steps of:

dissolving the compound 21 and 3, 4-dihydroxycinnamic acid in DMF, and carrying out condensation reaction in the presence of PyBOP and NMM to obtain the compound

3. Use of the sialylmethylester methyl glycoside derivative according to claim 1 for the preparation of a medicament for the prophylaxis or treatment of a neurological disease.

4. Use according to claim 3, characterized in that: the nervous system disease is neurodegenerative disease or cerebral ischemia.