CN103254120B

CN103254120B - Spiro-azasugar quaternary ammonium salt compound, and preparation method and application thereof

Info

Publication number: CN103254120B
Application number: CN201310182051.6A
Authority: CN
Inventors: 俞初一; 胡祥国; 贾月梅; 江新涛; 余官能; 章晓炜; 杨锦飞; 邱曾烨; 赵文博
Original assignee: Institute of Chemistry CAS
Current assignee: Institute of Chemistry CAS
Priority date: 2013-05-16
Filing date: 2013-05-16
Publication date: 2015-05-27
Anticipated expiration: 2033-05-16
Also published as: CN103254120A

Abstract

The invention discloses a kind of spiro-azasugar quaternary ammonium salt compounds and the preparation method and application thereof. The structure of the spiro-azasugar quaternary ammonium salt compound is shown in formula I. The preparation method of spiro-azasugar quaternary ammonium salt compound provided by the present invention is simple, and spiro-azasugar quaternary ammonium salt compound provided by the present invention can be used as glycosidase inhibitor, prevention and/or drug, prevention and/or drug, prevention and/or the drug and antiviral drugs for the treatment of tumour for the treatment of gaucher's disease for the treatment of diabetes. Formulas I.

Description

A kind of spiro-azasugar quaternary ammonium salt compound and preparation method thereof and application

Technical field

The present invention relates to a kind of spiro-azasugar quaternary ammonium salt compound and preparation method thereof and application.

Background technology

Azasugar is also known as iminosugar, polyhydroxylated alkaloid, imines sugar, imino-cyclitol, and it structurally can regard the analogue of monose as, the product that the Sauerstoffatom namely on sugared ring be instead of by nitrogen-atoms.So far, people have isolated from nature and have planted this kind of azasugar alkaloid more than more than 200.Azasugar causes the interest of people at first as glycosidase inhibitor, along with deepening continuously of research, except glucoside inhibiting activity, people have found again the inhibit activities of this compounds to other and sugared involved enzyme, as: glycosyltransferase, Glycofurol Starch phosphorylase, riboside dystopy enzyme.Based on above activity, azasugar can be used for treating diabetes, gaucher's disease, virus disease, tumour etc., is thus paid close attention to widely in new drug initiative field.Up to now, multiple Azasugar compounds has been had to be applied in some important diseases for the treatment of, such as, marketed drug miglitol (miglitol, type ii diabetes), NB-DNJ (Miglustat, gaucher's disease), nojirimycin hydrochloride (Pompeii sick) and be still in the Forodesine(leukemia of clinical experimental stage, BioCryst company), BCX-4208/R3421(psoriasis, Roche Holding Ag) and celgosivir(hepatitis C, Migenix company) etc.

Current separation obtains azasugar can be divided into five classes, that is: poly-hydroxy piperidines (Piperidine) alkaloid (as NJ), polyhydroxy pyrrole (Pyrrolidine) alkaloid (as DMDP), poly-hydroxy double pyrrolizidine alkane (Pyrrolizidine) alkaloid (as Alexine), poly-hydroxy Indolizidine (Indolizidine) alkaloid (as castanosperimine) and poly-hydroxy nortropane (Nortropane) alkaloid are (as Calystegine A ₃).In addition, people have also synthesized the derivative of many azasugars.Although people have synthesized some Azasugar compounds, the new Azasugar compound obtaining more good activity has been appointed so of crucial importance.

Summary of the invention

The object of the present invention is to provide a kind of new spiro-azasugar quaternary ammonium salt compound and preparation method thereof and application.

The invention provides a kind of spiro-azasugar quaternary ammonium salt compound, wherein, the compound that described volution azasugar quaternary ammonium compound is structure shown in formula I;

formula I

In formula I, R ¹and R ²independently of one another for hydrogen, phenyl, benzyl, hydrogen atom on phenyl ring by hydroxyl, carbonatoms be the alkoxyl group of 1-4, any one in the ether of at least one in nitro and the halogen benzyl, the carbonatoms that replace to be the acyl group of 2-10 and carbonatoms be 2-10, propylidene, benzal and cyclohexylidene; R ³and R ⁴independently of one another for hydrogen, benzyloxymethyl, methylol and carbonatoms are any one in the alkyl of 1-4; X is at least one in fluorion, chlorion, bromide anion, iodide ion, sulfonate ion, sulfate ion, hydrogen sulfate ion, nitrate ion, citrate ion, lactate ion, tartrate ion, phosphate anion, carbanion and bicarbonate ion; N is the integer of 1-3; Two m are identical or different, and be the integer of 0 or 1-3 separately, Y is CH ₂, NH, O or S.

The present invention also provides the preparation method of above-mentioned spiro-azasugar quaternary ammonium salt compound, and wherein, the method comprises the steps:

1) in the basic conditions, the dihalo thing of structure shown in the compound of structure shown in formula II and formula V is reacted, obtains the compound of structure shown in formula I;

formula II formula V

2) compound of structure shown in formula I and Lewis acid or hydroborating reagent are carried out deprotection reaction, obtain the compound of structure shown in formula Ia;

formula Ia

Wherein, R ¹and R ²independently of one another for phenyl, benzyl, hydrogen atom on phenyl ring by hydroxyl, carbonatoms be the alkoxyl group of 1-4, any one in the ether of at least one in nitro and the halogen benzyl, the carbonatoms that replace to be the acyl group of 2-10 and carbonatoms be 2-10, propylidene, benzal and cyclohexylidene; R ³and R ⁴independently of one another for hydrogen, benzyloxymethyl, methylol and carbonatoms are any one in the alkyl of 1-4; X is fluorine atom, chlorine atom, bromine atoms or atomic iodine; N is the integer of 1-3; Two m are identical or different, and be the integer of 0 or 1-3 separately, Y is CH ₂, NH, O or S.

The present invention also provides above-mentioned spiro-azasugar quaternary ammonium salt compound or its pharmaceutically acceptable hydrate or medicinal composition preparing glycosidase inhibitor, prevents and/or treats the medicine of diabetes, prevents and/or treats the medicine of gaucher's disease, the application prevented and/or treated in the medicine of tumour and antiviral.

The present invention also provides a kind of medicine, and wherein, the activeconstituents of this medicine contains above-mentioned spiro-azasugar quaternary ammonium salt compound or its pharmaceutically acceptable hydrate or medicinal composition.

According to the present invention, its preparation method is simple, and the spiro-azasugar quaternary ammonium salt compound obtained shows good inhibit activities to aftermentioned various Glycosylase.

Accompanying drawing explanation

Fig. 1 is the chemical reaction flow process figure of preparation formula II-1 compound.

Fig. 2 is the chemical reaction flow process figure of preparation formula II-5 compound.

Fig. 3 is the chemical reaction flow process figure of preparation formula II-6 compound.

Fig. 4 is the chemical reaction flow process figure of preparation formula II-7 compound.

Fig. 5 is the chemical reaction flow process figure of preparation formula I-1 compound.

Embodiment

Below the specific embodiment of the present invention is described in detail.Should be understood that, embodiment described herein, only for instruction and explanation of the present invention, is not limited to the present invention.

formula I

Carbonatoms is that the Alkoxy of 1-4 is as being methoxyl group, oxyethyl group, positive propoxy, isopropoxy, n-butoxy, isobutoxy or tert.-butoxy (following identical).

Halogen can be such as fluorine, chlorine, bromine or iodine (following identical).

Carbonatoms is the acyl group of 2-10 can be such as ethanoyl, propionyl, butyryl radicals, pentanoyl, caproyl, oenanthyl, capryloyl, nonanoyl or decanoyl (following identical).

Carbonatoms is the ether of 2-10 can be such as methoxyl methyl, methoxyethyl, ethoxymethyl, ethoxyethyl, isopropoxyethyl, benzyloxymethyl, tertiary fourth oxygen methyl or tri-chloroethoxy methyl (following identical).

Carbonatoms is the alkyl of 1-4 can be such as methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-or the tertiary butyl (following identical).

Preferably, in described formula I, R ¹and R ²be any one in hydrogen, benzyl, phenyl, benzoyl, ethanoyl, methoxyl methyl, ethoxymethyl and propylidene independently of one another; More preferably, R ¹and R ²be any one in hydrogen, benzyl and ethanoyl independently of one another; Further preferably, R ¹and R ²for hydrogen.

Preferably, in described formula I, R ³and R ⁴independently of one another for hydrogen, methylol and carbonatoms are any one in the saturated alkyl of 1-4; More preferably, R ³and R ⁴for hydrogen or methylol.

Preferably, in described formula I, X is fluorion, chlorion, bromide anion, iodide ion or sulfonate ion; More preferably, X is fluorion, chlorion, bromide anion or iodide ion.

Preferably, in described formula I, Y is CH ₂, NH or O.

In the present invention, spiro-azasugar quaternary ammonium salt compound shown in described formula I is particularly preferably any one in following formula I-1 ~ formula I-27:

Formula I-1 formula I-2 formula I-3 formula I-4 formula I-5 formula I-6 formula I-7

Formula I-8 formula I-9 formula I-10 formula I-11 formula I-12 formula I-13 formula I-14

Formula I-15 formula I-16 formula I-17 formula I-18 formula I-19 formula I-20 formula I-21

Formula I-22 formula I-23 formula I-24 formula I-25 formula I-26 formula I-27.

The present invention also provides the preparation method of above-mentioned spiro-azasugar quaternary ammonium salt compound, and the method comprises the steps,

formula II formula V

formula Ia

R ¹and R ²independently of one another for phenyl, benzyl, hydrogen atom on phenyl ring by hydroxyl, carbonatoms be the alkoxyl group of 1-4, any one in the ether of at least one in nitro and the halogen benzyl, the carbonatoms that replace to be the acyl group of 2-10 and carbonatoms be 2-10, propylidene, benzal and cyclohexylidene; R ³and R ⁴independently of one another for hydrogen, benzyloxymethyl, methylol and carbonatoms are any one in the alkyl of 1-4; X is fluorine atom, chlorine atom, bromine atoms or atomic iodine; N is the integer of 1-3; Two m are identical or different, and be the integer of 0 or 1-3 separately, Y is CH ₂, NH, O or S.

The steric configuration of all compounds of the present invention does not do clear stipulaties, and different steric isomers can be prepared as starting raw material by selecting the monose of respective configuration.

In above-mentioned steps 1) in, by the basic conditions, the dihalo thing of structure shown in the compound of structure shown in formula II and formula V is reacted, obtains the compound of structure shown in formula I.

Preferred steps 1) reaction carry out under solvent, all kinds of SOLVENTS that described solvent commonly uses by this area can be such as organic solvent, the mixed solvent of organic solvent or the mixed solvent etc. of organic solvent and water.Under preferable case, described organic solvent is one or more in acetonitrile, toluene, tetrahydrofuran (THF), Isosorbide-5-Nitrae-dioxane and dimethyl formamide; Be more preferably acetonitrile.

Above-mentioned steps 1) in alkaline condition, by adding alkali to realize in reaction system.Described alkali can the various organic bases commonly used by this area or mineral alkali.But in preferred situation, described alkali is one or more in triethylamine, diisopropyl ethyl amine, Tributylamine, sodium carbonate, sodium bicarbonate and salt of wormwood.Be more preferably salt of wormwood.

Above-mentioned steps 1) in, shown in the compound of structure shown in formula II, formula V, the dihalo thing of structure and the mol ratio of above-mentioned alkali are 1:1-100:1-100; Be preferably 1:2-5:5-10.

Under preferable case, above-mentioned steps 1) reaction conditions comprise: temperature of reaction is 0-100 DEG C, and the reaction times is 0.5-72 hour; Being more preferably temperature of reaction is 60-80 DEG C, and the reaction times is 0.5-24 hour.

In addition, for above-mentioned steps 1) reacted purifying, various method known in the field can be adopted carry out, thus obtain the compound of structure shown in formula I.

According to the present invention, in the compound of structure shown in the formula I obtained by aforesaid method, R ¹and R ²independently of one another for phenyl, benzyl, hydrogen atom on phenyl ring by hydroxyl, carbonatoms be the alkoxyl group of 1-4, any one in the ether of at least one in nitro and the halogen benzyl, the carbonatoms that replace to be the acyl group of 2-10 and carbonatoms be 2-10, propylidene, benzal and cyclohexylidene; R ³and R ⁴independently of one another for hydrogen, benzyloxymethyl, methylol and carbonatoms are any one in the alkyl of 1-4; N is the integer of 1-3; X is fluorine atom, chlorine atom, bromine atoms or atomic iodine; Two m are identical or different, and be the integer of 0 or 1-3 separately, Y is CH2, NH, O or S.

Preferably, in the compound of structure shown in the formula I obtained by aforesaid method, R ¹and R ²be any one in benzyl, phenyl, benzoyl, ethanoyl, methoxyl methyl, ethoxymethyl and propylidene independently of one another; More preferably, R ¹and R ²be benzoyl, benzyl or ethanoyl independently of one another.

Preferably, in the compound of structure shown in the formula I obtained by aforesaid method, R ³and R ⁴independently of one another for hydrogen, benzyloxymethyl, methylol and carbonatoms are any one in the saturated alkyl of 1-4; More preferably, R ³and R ⁴for hydrogen, methylol or benzyloxymethyl.The saturated alkyl being 1-4 as carbonatoms is methyl, ethyl, n-propyl and normal-butyl

According to the present invention, in above-mentioned steps 2) in, the compound of structure shown in formula I obtained above and Lewis acid or hydroborating reagent are carried out deprotection reaction, obtains the compound of structure shown in formula Ia.

Above-mentioned Lewis acid can the various Lewis acids commonly used by this area.Under preferable case, described Lewis acid is one or more in boron tribromide, boron trichloride and boron trifluoride; Be more preferably boron trichloride and/or boron tribromide.

Above-mentioned hydroborating reagent also can the various hydroborating reagents commonly used by this area.Under preferable case, described hydroborating reagent is Pd/C/H ₂, Pd (OH) ₂/ H ₂, palladium black/H ₂with platinum/H ₂in one or more; Be more preferably Pd/C/H ₂.

Above-mentioned steps 2) in, the compound of structure shown in formula I and lewis acidic mol ratio are 1:1-50; Be preferably 1:1-10; Most preferably be 1:5.

Above-mentioned steps 2) in, the compound of structure shown in formula I and the mass ratio of hydroborating reagent are 1:0.1-10, are preferably 1:0.05-0.1.

Above-mentioned steps 2) in, the condition of described deprotection reaction comprises: temperature of reaction is 0-100 DEG C, and the reaction times is 0.5-72 hour; Being preferably temperature of reaction is 0-40 DEG C, and the reaction times is 0.5-24 hour.

In addition, for the purifying after above-mentioned deprotection reaction, various method known in the field can be adopted carry out, thus obtain the compound of structure shown in formula Ia.

According to the present invention, in the compound of structure shown in the formula Ia obtained by aforesaid method, R ³and R ⁴independently of one another for hydrogen, benzyloxymethyl, methylol and carbonatoms are any one in the alkyl of 1-4; N is the integer of 1-3; X is fluorine atom, chlorine atom, bromine atoms or atomic iodine; Two m are identical or different, and be the integer of 0 or 1-3 separately, Y is CH2, NH, O or S.

Preferably, in the compound of structure shown in the formula Ia obtained by aforesaid method, R ³and R ⁴independently of one another for hydrogen, benzyloxymethyl, methylol and carbonatoms are any one in the saturated alkyl of 1-4; More preferably, R ³and R ⁴for hydrogen or methylol.The saturated alkyl being 1-4 as carbonatoms is methyl, ethyl, n-propyl and normal-butyl.

According to the present invention, the spiro-azasugar quaternary ammonium salt compound obtained by aforesaid method be particularly preferably in compound shown in above-mentioned formula I-1 ~ formula I-27 any one.

In addition, be the Azasugar quaternary ammonium salt compound of sulfate ion, hydrogen sulfate ion, nitrate ion, citrate ion, lactate ion, tartrate ion, phosphate anion, carbanion or bicarbonate ion to obtain X in formula 1, shown in the formula I that corresponding acid and aforesaid method can be adopted to obtain or formula Ia, compound carries out permutoid reaction, and permutoid reaction condition is well known in the art.

In addition, the compound (i.e. the secondary amine of polyhydroxy annular) of structure shown in the starting materials II that the present invention is used, can be prepared by corresponding sugar, conventional sugar has wood sugar, pectinose, ribose, glucose, seminose, semi-lactosi etc., its preparation method can with reference to following patent and document [(a) Overkleeft, H.S. Tetrahedron1994 is waited, 50,4215-4224; A kind of (b) method preparing polyhydroxy annular nitrone, ZL200610066638.0; The Synlett such as (c) Wang W.B., 2010,3,488-492; (d) Matos, the Synthesis such as C.R., 1999,571-573; (e) Rountree, the Tetrahedron Letters such as J.S., 2007,48,24,4287-4291]

Specifically, the compound of structure shown in above-mentioned formula II includes but not limited to following arbitrary structure

According to the present invention, above-mentioned spiro-azasugar quaternary ammonium salt compound or its pharmaceutically acceptable hydrate or medicinal composition also belong to protection scope of the present invention.

Above-mentioned spiro-azasugar quaternary ammonium salt compound provided by the present invention or its pharmaceutically acceptable hydrate or medicinal composition are particularly preferred for preparing glycosidase inhibitor.

Described Glycosylase can be at least one in alpha-glucosidase, beta-glucosidase, alpha-galactosidase, beta-galactosidase enzymes, alpha-Mannosidase, beta-Mannosidase, alpha-L-fucosidase, beta-glucuronidase, α, α-trehalase and alpha-L-Rhamnosidase.

Above-mentioned spiro-azasugar quaternary ammonium salt compound provided by the present invention or its pharmaceutically acceptable hydrate or medicinal composition can also as activeconstituents for the preparation of any one medicines following: the medicine 1) preventing and/or treating diabetes; 2) medicine of gaucher's disease is prevented and/or treated; 3) medicine of tumour is prevented and/or treated; 4) antiviral.

According to the present invention, in said medicine, be 1-99% weight as the provided by the present invention above-mentioned spiro-azasugar quaternary ammonium salt compound of activeconstituents or the content of its pharmaceutically acceptable hydrate or medicinal composition; Be preferably 10-90 % by weight.

In the present invention, as required, one or more pharmaceutically acceptable carriers can also can also be added in said medicine.Described carrier comprises the thinner of pharmaceutical field routine, vehicle, weighting agent, tackiness agent, wetting agent, disintegrating agent, absorption enhancer, tensio-active agent, absorption carrier, lubricant and other optional additives.The medicine prepared with above-mentioned spiro-azasugar quaternary ammonium salt compound or its pharmaceutically acceptable hydrate or medicinal composition can be the various ways such as injection liquid, tablet, pulvis, granule, capsule, oral liquid, paste, creme.The medicine of above-mentioned various formulation all can be prepared according to the ordinary method of pharmaceutical field.

Described medicine can utilize various route of administration administration, includes but not limited to oral, suction, rectum, transdermal, administration in mucous membrane intestines, and subcutaneous, muscle or intravenous administration.Above-mentioned spiro-azasugar quaternary ammonium salt compound of the present invention or its pharmaceutically acceptable hydrate or medicinal composition, can be individually dosed, or with other known treating diabetes, antiviral, antibacterial administration together with antitumor drug.

Spiro-azasugar quaternary ammonium salt compound provided by the invention, through the test of Glycosylase inhibition, proves that the various Glycosylases to using in aftermentioned embodiment 28 show good inhibit activities.

Below in conjunction with specific embodiment, the present invention is further elaborated, but the present invention is not limited to following examples.Described method is ordinary method if no special instructions.Described starting material all can obtain from open commercial sources if no special instructions.The steric configuration of all compounds of the present invention does not do clear stipulaties, is configured as R or S when relating to chiral centre, and different steric isomers can be prepared as starting raw material by selecting the monose of respective configuration.

Compound shown in raw materials used in following embodiment (II-1)-(II-8) is [(a) Overkleeft that the method provided according to following patent documentation and non-patent literature carries out synthesizing, H.S. Tetrahedron1994 is waited, 50,4215-4224; A kind of (b) method preparing polyhydroxy annular nitrone, ZL200610066638.0; The Synlett such as (c) Wang W.B., 2010,3,488-492; (d) Matos, the Synthesis such as C.R., 1999,571-573; (e) Rountree, the TetrahedronLetters such as J.S., 2007,48,24,4287-4291].

Wherein, the Bn in (II-1)-(II-8) is benzyl, and MOM is methoxyl methyl (-CH2OMe).Illustrate: document a is the preparation for II-1, II-2, II-3 and II-4; II-2 also can be prepared according to document d; Document e is used for the preparation of II-5; Document b, c are used for the preparation of II-6.

Concrete steps prepared by each raw material are as follows:

The preparation (reference (a)) of II-2:

By 2 under normal temperature, 3, 4, 6-tetra--O-benzyl-D-MANNOSE (being purchased from Beijing Kai Senlai Pharmaceutical Technology Co., Ltd) II-2-1(75.5g, 0.140mol) be dissolved in diacetyl oxide (211mL), and add dimethyl sulfoxide (DMSO) (238mL) wherein, stirring at normal temperature reaction 12h under argon shield, TLC adds water (800mL) after monitoring raw material disappearance will react cancellation, leave standstill after stirring 15min, branch vibration layer, oil reservoir is with mixed solvent (ethyl acetate/petroleum ether=1:1(volume ratio)) dissolve, wash three times, the above-mentioned mixed extractant solvent of aqueous phase, merge organic phase, anhydrous magnesium sulfate drying, concentrated, recrystallization from ethyl acetate/petroleum ether, obtain 49.8g white solid II-2-2, mother liquor is separated to obtain product 2g through chromatography column, productive rate 72%.

By tetrabenzyl seminose lactone II-2-2(47g, 0.087mol under normal temperature) be dissolved in saturated methanolic ammonia solution (700mL), normal-temperature reaction 3h under argon shield.TLC monitors after raw material disappears and filters out white solid product, filter cake petroleum ether, adds sherwood oil and ethyl acetate mixed solvent (petrol ether/ethyl acetate=3/1(volume ratio) after filtrate is concentrated), stir, filter, collect filter cake, obtain 43.7g white solid II-2-3, productive rate 91%.

By acid amides II-2-3(48.7g under normal temperature, 0.088mol) be dissolved in diacetyl oxide (156mL), add methyl-sulphoxide (263mL), under argon shield, stirring at normal temperature reaction 20h, TLC adds water (300mL) after monitoring raw material disappearance will react cancellation, leave standstill after stirring 15min, filter, filter cake sherwood oil and ethyl acetate mixed solvent (petrol ether/ethyl acetate=3/1) washing, collect filter cake, filtrate is again with mixed solvent ethyl acetate/petroleum ether=1:1 extraction, organic phase washed with water is washed till without methyl-sulphoxide, again through sherwood oil and ethyl acetate mixed solvent recrystallization after concentrated, obtain 47g white solid II-2-4, productive rate 96%.

By 5-oxanamide II-2-4(47g, 0.085mol) be dissolved in saturated methanolic ammonia solution (700mL), the lower 30 DEG C of reaction 4h of argon shield.After TLC monitors raw material disappearance, after reaction solution is concentrated, obtain II-2-5, be dissolved in acetonitrile (500mL) with the solvent mixture of formic acid (282mL), then add sodium cyanoborohydride (16.9g, 0.268mol), the lower 70 DEG C of back flow reaction 3h of argon shield in batches.After TLC monitors raw material disappearance, add the hydrochloric acid soln (30mL) of 0.1mol/L, stir and add ethyl acetate again after 15min and saturated sodium bicarbonate solution is neutralized to alkalescence, separatory, aqueous phase is extracted with ethyl acetate, and merges organic phase, anhydrous magnesium sulfate drying, 41.6g pale yellowish oil product II-2-6 is obtained, productive rate 91% after concentrated.

By lactan II-2-6(43.3g, 0.081mol) be dissolved in tetrahydrofuran (THF) (700mL), slowly add lithium aluminum hydride (8.76g, 0.230mol), under argon shield, 70 DEG C of return stirring reaction 8h.TLC adds ethyl acetate after monitoring raw material disappearance will react cancellation, the sodium hydroxide solution adding 10% more no longer increases to White Flocculus in system, filters, filtrate organic phases washed with water three times, aqueous phase is extracted with ethyl acetate, merge organic phase, organic phase saturated common salt is washed once, anhydrous magnesium sulfate drying, concentrated, column chromatography purification (silica gel, petrol ether/ethyl acetate=5/1(volume ratio)+0.5% triethylamine) obtain product II-2(16.8g, productive rate 39%).

¹HNMR（300MHz，CDCl ₃）：δ7.39-7.19（m，20H），4.93（d，J=10.8Hz，1H），4.75-4.43（m，7H），3.93（t，J=9.6Hz，1H），3.77-3.71（m，2H），3.62-3.59（m，1H），3.49（dd，J=2.7，9.3Hz，1H），3.13（d，J=14.1Hz，1H），2.61-2.58（m，1H），2.48（d，J=14.4Hz，1H），2.13（bs，1H）； ¹³CNMR（75MHz，CDCl ₃）：δ138.95，138.89，138.75，138.49，128.71，128.60，128.47，128.43，128.18，128.02，127.97，127.77，127.64，127.47，84.51，76.96，75.44，73.88，73.34，71.82，71.30，69.69，60.39，46.91。

The preparation (reference (a)) of II-1:

Carry out according to the method for the above-mentioned II-2 of preparation, unlike by raw material 2,3,4,6-tetra--O-benzyl-D-MANNOSE changes 2 into, 3,4,6-tetra--O-benzyl-D-Glucose (being purchased from Beijing Ribio Biotech. Inc.), prepares II-1(nuclear magnetic data as follows); In addition, II-1 also can be prepared by reference (d).

¹H-NMR（300MHz，CDCl ₃）：δ2.05（1H，br，s），2.50（1H，dd，J=10.1，12.2Hz），2.71（1H，ddd，J=2.6，5.9，9.7Hz），3.25（1H，dd，J=4.7，12.2Hz），3.35（1H，t，J=11.8Hz），4.47（1H，d，J=11.8Hz），4.49（1H，d，J=10.9Hz），4.66（1H，d，J=11.7Hz），4.70（1H，d，J=11.7Hz），4.83（1H，d，J=10.9Hz），4.86（1H，d，J=10.9Hz），4.98（1H，d，J=10.9Hz），7.30（20H，m）； ¹³C-NMR（75MHz，CDCl ₃）：δ49.29，59.90，70.42，72.97，73.57，75.38，75.6，80.25，80.82，87.50，127.86，127.97，128.05，128.14，128.20，128.57，128.59，138.15，138.57，138.67，139.07。

The preparation of II-3: according to the preparation method of above-mentioned II-2, changes 2,3,4,6-tetra--O-benzyl-D-semi-lactosi (being purchased from Beijing Ribio Biotech. Inc.) into unlike by raw material 2,3,4,6-tetra--O-benzyl-D-MANNOSE, prepares II-3.

¹HNMR（300MHz，CDCl ₃）：δ7.40-7.23（m，20H），4.94（d，J=9.0Hz，1H），4.77-4.39（m，7H），3.96（bs，1H），3.88（td，J=5.4Hz，9.9Hz，1H），3.50-3.44（m，2H），3.36-3.23（m，2H），2.78（t，J=6.6Hz，1H），2.47（dd，J=10.5Hz，12.6Hz，1H），1.87（bs，1H）； ¹³CNMR（75MHz，CDCl ₃）：δ139.01，138.98，138.10，128.54，128.46，128.43，128.33，128.21，128.05，127.86，127.68，127.61，127.58，85.01，76.85，74.95，74.54，73.54，73.21，72.72，70.30，58.61，49.19。

The preparation of II-4: according to the preparation method of above-mentioned II-5, changes D-R (being purchased from Beijing Ribio Biotech. Inc.) into unlike by raw material D-wood sugar, prepares II-4.

¹HNMR（300MHz，DCCl ₃）：δ7.33-7.24（m，15H），4.58-4.36（m，6H），4.07（s，1H），4.98（s，1H），3.78-3.71（m，3H），3.49-3.46（m，1H），3.36（dd，J=12.6，4.5Hz）； ¹³CNMR（75MHz，DCCl ₃）：47.4，61.9，66.7，70.5，70.9，72.3，75.7，76.2，76.6，79.7，81.4，126.8，126.9，127.0，127.4，127.5，136.1，136.2，136.6。

The preparation (reference (e)) of II-5:

Get 2L flask, add 1L methyl alcohol, dropwise drip Acetyl Chloride 98Min. (20mL) under ice bath agitation condition, then add D-wood sugar (100g, 666mmol), keep zero degree reaction, until raw material disappears, be neutralized to neutrality with sodium bicarbonate, remove inorganic salt with diatomite filtration, filtrate concentrates under vacuum, obtains crude pale yellow oil shape thing and directly casts single step reaction.

Previous step crude product (calculating according to 666mmol) is dissolved into dry DMF(200mL) in solution, and be dropwise added drop-wise to NaH(123g, 2.80mol, content 60%), in tetrahydrofuran (THF) (800mL) and mixed solution DMF(600mL), finish, add TBAI(tetrabutylammonium iodide, 4.07g), after half an hour, drip BnBr(2.53mol, 331.6mL), reaction is until raw material disappears, slowly add ice cube and extract reaction of going out, be separated organic phase, aqueous phase is extracted with ethyl acetate (3 × 300mL), merge organic phase, washing organic phase removes DMF, organic phase anhydrous magnesium sulfate drying, cross and filter siccative, filtrate concentrates to obtain crude product under vacuum, directly cast single step reaction

Upper step crude product (calculating by 666mmol) is dissolved into glacial acetic acid aqueous solution (800mL, volume content 80%) in, add hydrochloric acid (1mol/L again, 133mL), control temperature 95 DEG C reaction is until raw material disappears, add saturated sodium bicarbonate solution and extract reaction of going out, be separated organic phase, aqueous phase is extracted with ethyl acetate (3 × 300mL), merge organic phase, after use dried over mgso organic phase, cross and filter siccative, filtrate concentrates under vacuum, by column chromatography for separation (ethyl acetate/petroleum ether=1:10(volume ratio)) obtain 196g yellow oil II-5-1, productive rate is added up to be 70%.

By sodium borohydride (50.7g, 666mmol) join in the dichloromethane solution (200mL) of above-mentioned product II-5-1, then methyl alcohol (800mL) is dropwise dripped, stir until raw material disappears, the extraction that adds water is gone out reaction, be separated organic phase, aqueous phase is extracted with ethyl acetate (3 × 300mL), merge organic phase, anhydrous magnesium sulfate drying, cross and filter siccative, filtrate concentrates under vacuum, by column chromatography for separation (ethyl acetate/petroleum ether=1:3(volume ratio)) obtain 114.9g light yellow oil II-5-2, yield is 76%.

Get 500mL there-necked flask, by triethylamine (7.17g, 70mmol, 9.9mL) add in the dichloromethane solution (250mL) of II-5-2, under 0 DEG C of condition, drip methylsulfonyl chloride (6.54g, 57mmol, 4.4mL), TLC monitoring reaction is until raw material disappears, then the extraction that adds water is gone out reaction, aqueous phase ethyl acetate (3 × 50mL) extracts, merge organic phase, anhydrous magnesium sulfate drying, cross and filter siccative, filtrate concentrates under vacuum, by column chromatography (ethyl acetate/petroleum ether=1:3(volume ratio)) separation and purification, obtain 38.8g light yellow oil II-5-3, yield 96%.

By II-5-3(31.5g) add in the reaction flask that benzylamine (40mL) is housed, stir under oil bath computer heating control temperature 60 C, TLC monitoring reaction, until raw material disappears, drips hydrochloric acid (3mol/L, 40mL), aqueous phase methylene dichloride (3 × 30mL) extraction, merges organic phase, anhydrous magnesium sulfate drying, cross and filter siccative, filtrate concentrates under vacuum, by column chromatography (ethyl acetate/petroleum ether=1:20(volume ratio)) be separated to obtain 19.6g yellow oil II-5-4, yield 73%.

By II-5-4(19.6g, 43mmol) be dissolved in and be equipped with 1, in the reaction flask of 2-ethylene dichloride (75mL), then ethyl chloroformate (10.3g is dripped, 72mmol, 7.5mL), oil bath refluxes under heating 100 DEG C of conditions, and TLC monitoring is reacted until raw material disappears, solvent evaporated, add methyl alcohol (75mL), continue to reflux under oil bath is heated to 70 ° of C conditions, TLC monitoring is reacted until raw material disappears, solvent evaporated, through column chromatography (methylene chloride/methanol=60:1(volume ratio)) separation and purification obtains 13.7g atropurpureus oily liquids II-5, yield 86%.Its nuclear magnetic data is: ¹hNMR(300MHz, CDCl ₃): δ 7.33-7.24(m, 15H), 4.58-4.36(m, 6H) and, 4.07(s, 1H), 4.98(s, 1H) and, 3.78-3.71(m, 3H), 3.49-3.46(m, 1H) and, 3.36(dd, J=12.6,4.5Hz); ¹³cNMR(75MHz, CDCl ₃): δ 136.6,136.2,136.1,127.5,127.4,127.0,126.9,126.8,81.4,79.7,76.6,76.2,75.7,72.3,70.9,70.5,66.7,61.9,47.4.

The preparation (reference (b)) of II-6:

Get II-6-2(according to document organic synthesis coll.Vol.1998,9,704. preparations, 26g, 71mmol) be placed in two mouthfuls of bottles, with air in argon replaces bottle three times, (newly heavily steam through sodium backflow, benzophenone colour developing becomes blue to get dry tetrahydrofuran solution with syringe, 80mL) inject the reaction flask having finished changing argon gas-sealed, reaction flask is placed in-80 DEG C of low temperature alcohol bath mortises, n-Butyl Lithium (the 27mL of 2.5M is dripped after half an hour, 69mmol), control temperature in bottle and be less than-60 DEG C, dripped more than 20 minutes, by nitrone II-6-1(9.9g after 15 minutes, 23.6mmol) be dissolved in dry tetrahydrofuran solution (20mL) and be slowly injected in reaction flask, continue stirring after 30 minutes, reaction flask is placed in the cancellation of room temperature saturated aqueous ammonium chloride, add ethyl acetate (100mL) and water (100mL) and stratification, by ethyl acetate (300mL) point three extracted organic phase, merge organic phase and use anhydrous magnesium sulfate drying, concentrating under reduced pressure, 5.1g light yellow liquid II-6-3 is obtained through purification by column chromatography, productive rate 44%.

Zinc powder (the 4.5g activated is added in Glacial acetic acid (30mL), 67.8mmol) and neutralized verdigris (280mg, 1.4mmol), under stirring at room temperature state and suitable electric heat gun is heated to system is brown, azanol II-6-3(3.34g is dissolved with methylene dichloride (20mL) after 15 minutes, 6.78mmol) and join reaction system, stirring at room temperature is continued.After half an hour, TLC display raw material reaction is complete.Directly by reaction flask vacuum concentration, and use dchloromethane suction filtration, water (50mL) is added in mother liquor, regulate mother liquor pH value to being greater than 10 with 6N sodium hydroxide, stratification, by methylene dichloride (150mL) point three extracted organic phase, merge organic phase and use Magnesium Chloride Anhydrous dry, concentrating under reduced pressure solvent, obtains 3.23g orange II-6, productive rate 100%.Its specific rotation, infrared data, nuclear magnetic data are as follows: -12.5(c2.40, CH ₂cl ₂); IR(cm ^-1): 3030(w), 2868(m), 1496(w), 1454(m) and, 1363(w), 1208(w) and, 1103(s), 1043(m) and, 737(m), 698(m); ¹hNMR(300MHz, CDCl ₃): δ (ppm) 4.60(s, 2H), 4.54-4.50(m, 6H) and, 3.97-3.89(m, 2H), 3.64-3.52(m, 4H) and, 3.44-3.33(m, 2H), 3.31(s, 3H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 138.2,138.2,128.4,127.9,127.9,127.8,127.7,127.7,96.7,86.1,77.6,73.3,72.0,71.9,70.5,68.1,61.6,61.5,55.4.

The preparation of II-7: (the II-7-1), according to document [Ackermann, L. from PEARLITOL 25C; ElTom, D.; Furstner, A.Tetrahedron2000,56,2195.] method synthesis glycol II-7-4.PEARLITOL 25C (100.0g, 0.549mol) is dissolved in pyridine (600mL), add triphenylmethyl chloride (314g, 1.13mol) and 4-dimethylamino pyridine (1.34g, 11mmol), reaction mixture stirs 36 hours in room temperature.Be spin-dried for pyridine, add 1L ethyl acetate, washing (2 × 500mL), organic phase anhydrous magnesium sulfate drying.Filter, concentrated filtrate obtains the crude product of Compound II per-7-2, is directly used in next step reaction.

Be scattered in by sodium hydride (60%, 121g, 3.02mol) in tetrahydrofuran (THF) (800mL), dropwise drip the tetrahydrofuran solution (200mL) of the Compound II per-7-2 that above-mentioned steps obtains, stirring at room temperature about 2 is little of releasing without gas.Add tetrabutylammonium iodide (3.69g, 10mmol), and dropwise drip bromobenzyl (329mL, 2.75mol), in process, maintain the temperature at 40-45 ° of C.Drip off rear stirring 2 hours, add frozen water cancellation reaction.Be extracted with ethyl acetate (3 × 500mL), organic phase anhydrous magnesium sulfate drying.Filter, concentrated filtrate obtains the PEARLITOL 25C II-7-3 of hydroxyl full guard, and crude product is directly used in the next step.

Compound II per-7-3 is dissolved in methylene dichloride (500mL) and methyl alcohol (500mL), under ice-water bath, adds the vitriol oil (80mL), add rear room temperature reaction and spend the night.Add strong aqua and be adjusted to pH=9, concentration of reaction solution.Add water 1L, is extracted with ethyl acetate (3 × 800mL).Organic phase anhydrous magnesium sulfate drying.Filter, concentrated filtrate, carries out column chromatography (petrol ether/ethyl acetate=4:1(volume ratio) by resistates), obtain glycol II-7-4(162.0g, three step overall yields 54%), yellow oil.

Under ice-water bath, methane sulfonyl chloride (460mg, 4mmol) is dropwise added dropwise to glycol II-7-4(543mg, 1.0mmol) and triethylamine (606mg, 6.0mmol) methylene dichloride (5mL) solution in, add rear stirring at room temperature 1 hour.Reaction solution is diluted to 50mL, washing (2 × 10mL).Organic phase anhydrous sodium sulfate drying, filters and concentrated filtrate.Resistates is dissolved in allyl amine (5mL), refluxes 24 hours.Concentrated, post is separated (petrol ether/ethyl acetate=5:1(volume ratio)) obtain tertiary amine II-7-5(398mg, 2 step productive rates 71%), colorless oil.

To tertiary amine II-7-5(605mg, 1.07mmol) 1,2-ethylene dichloride (5mL) in add chloroformic acid 1-chloroethene ester (307mg, 2.14mmol), backflow is spent the night.Concentration of reaction solution, is dissolved in methyl alcohol (5mL) by resistates, backflow is spent the night again.Concentration of reaction solution, is separated (methylene chloride/methanol=100:1(volume ratio) by residue post), obtain azepan II-7(480mg, 86%), white solid.Its each test data is as follows: m.p.76-78 ° of C; [α] _d ²⁰=-34.0(c1.0, CHCl ₃); IR(cm ^-1): 3444,3062,3031,2919,1589,1496,1454,1369,1207,1095,1069,1027,909,736,698; ¹hNMR(300MHz, CDCl ₃): δ 7.36-7.16(m, 20H), 4.74-4.49(m, 8H) and, 4.09(dd, J=1.2,8.4Hz, 2H), 3.72(s, 2H) and, 3.50-3.43(m, 2H), 3.36-3.32(m, 2H); ¹³cNMR(75MHz, CDCl ₃): δ 138.64,138.52,128.42,128.39,127.83,127.71,127.66,78.75,77.76,73.62,71.89,46.22; HRMS-ESI(m/z) calcd forC ₃₄h ₃₈nO ₄[M+H ⁺] 524.2795, found524.2790.

The preparation of II-8: carry out according to the preparation method of above-mentioned II-7, be changed to D-Glucose alcohol unlike by raw material PEARLITOL 25C, prepare II-8.

¹HNMR（300MHz，CDCl ₃）：δ7.29-7.18（m，20H），4.64-4.47（m，8H），3.94-3.87（m，2H），3.83（t，J=4.5Hz，1H），3.68-3.65（m，1H），3.36-3.24（m，2H），3.15（dd，J=4.5，13.2Hz，1H），2.99（dd，J=1.5，14.1Hz，1H）; ¹³CNMR（75MHz，CDCl ₃）:δ138.38，138.36，138.18，138.14，128.45，128.41，128.36，127.89，127.84，127.78，127.74，127.63，82.61，80.46，80.10，77.40，72.96，72.89，71.84，71.73，47.39，46.33。

Embodiment 1:(2R, 3R, 4R, 5S) preparation of-N, N-(tetramethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-1)

1) by (2R, 3R, 4R, 5S)-2-benzyloxymethyl-3, 4, 5-tri--benzyloxy piperidines II-1(502mg, 0.96mmol) be dissolved in dry acetonitrile (30mL), add salt of wormwood (927mg successively, 6.72mmol), 1, 4-dibromobutane (0.5mL, 4.2mmol), reflux 24 hours, when reacting completely (TLC monitoring), suction filtration, concentrated filtrate, column chromatography (methylene dichloride: methyl alcohol=60:1(volume ratio)) obtain white solid (2R, 3R, 4R, 5S)-N, N-(1, 4-butylidene)-2-benzyloxymethyl-3, 4, 5-tri--benzyloxy piperidinium bromide (IV-1) (386mg, 0.59mmol), productive rate is 61%.Its nuclear magnetic data is: ¹hNMR(300MHz, CDCl ₃): δ 7.37-7.15(m, 20H), 4.89-4.71(m, 4H) and, 4.62-4.41(m, 4H), 4.21-4.03(m, 5H) and, 3.99-3.84(m, 6H), 3.75-3.67(m, 1H) and, 2.30-2.16(m, 2H), 2.04-1.90(m, 2H); ¹³cNMR(75MHz, CDCl ₃): δ 137.5,137.3,137.2,136.5,128.7,128.5,128.5,128.4,128.3,128.1,128.1,128.0,127.9,127.5,79.0,77.3,75.0,74.6,73.6,73.4,73.1,70.6,67.2,65.0,61.9,59.9,22.9,22.7.

By (2R, 3R, 4R, 5S)-N, N-(1, 4-butylidene)-2-benzyloxymethyl-3, 4, 5-tri--benzyloxy piperidinium bromide (IV-1) (326mg, 0.49mmol) be dissolved in methyl alcohol (25mL), add 10%Pd/C(30mg), stir lower nitrogen replacement 3 times, react under a hydrogen atmosphere, after reacting completely (TLC monitoring), suction filtration, concentrated filtrate, strip with ether after water-soluble for residue, concentrated aqueous phase obtains colorless oil (2R, 3R, 4R, 5S)-N, N-(1, 4-butylidene)-2-methylol-3, 4, 5-trihydroxy-piperidinium bromide (I-1) (123mg, 0.55mmol), productive rate is 84%.Its nuclear magnetic data is: ¹hNMR(300MHz, D ₂o): δ 4.22(d, 1H, J=7.23Hz), 4.09-4.00(m, 2H), 3.91-3.84(m, 2H), 3.79(dd, 1H, J=9.67Hz, 10.81Hz), 3.62(dd, 1H, J=4.66Hz, 12.76Hz), 3.56-3.43(m, 4H), 3.21(t, 1H, J=12.30Hz), 2.18(m, 4H); ¹³cNMR(75MHz, D ₂o): δ 76.4,72.9,67.6,67.4,65.5,64.6,58.8,56.1,24.3,22.8.

Embodiment 2:(2R, 3R, 4R, 5S) preparation of-N, N-(pentamethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-2)

According to the method for embodiment 1 with II-1 and pentamethylene bromide for raw material prepares compound IV-2 and I-2.

IV-2： ¹HNMR（300MHz，CDCl ₃）：δ7.35-7.18（m，20H），4.81-4.45（m，8H），4.35（bs，1H），4.23-4.12（m，2H），4.12-4.93（m，5H），3.71-3.63（m，2H），3.59-3.55（m，1H），3.48-3.44（m，1H），1.80-1.24（m，6H）； ¹³CNMR（75MHz，CDCl ₃）：δ137.6，137.4，137.3，136.7，128.6，128.5，128.5，128.4，128.2，128.1，127.9，127.8，127.5，80.8，77.7，74.6，73.9，73.3，73.2，72.4，71.0，63.2，62.2，56.9，54.3，20.6，20.0，19.5。

I-2： ¹HNMR（300MHz，D ₂O）：δ4.29（d，1H，J=7.35Hz），4.21-4.12（m，2H），4.00-3.85（m，3H），3.56-3.43（m，3H），3.27（dd，1H，J=2.94Hz，10.47Hz），3.19（d，1H，J=6.04Hz），2.87（t，1H，J=12.60Hz），2.16-1.98（m，1H），1.97-1.76（m，4H），1.56-1.42（m，1H）； ¹³CNMR（75MHz，D ₂O）：δ76.8，75.8，65.8，63.8，62.7，56.1，54.8，54.0，20.5，19.5，18.8。

Embodiment 3:(2R, 3R, 4R, 5S) preparation of-N, N-(hexamethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-3)

Be that raw material prepares compound IV-3 and I-3 according to the method for embodiment 1 with II-1 and 1,6-dibromo-hexane.

IV-3： ¹HNMR（300MHz，CDCl ₃）：δ7.36-7.18（m，20H），4.93-4.59（m，6H），4.47-4.37（m，2H），4.32（s，1H），4.21-4.06（m，3H），4.00-3.95（m，3H），3.86（t，1H，J=10.90Hz），3.78-3.74（m，2H），3.56（dd，1H，J=3.78Hz，13.00Hz），3.23（dd，1H，J=6.94Hz，13.81Hz），1.70-1.26（m，8H）； ¹³CNMR（75MHz，CDCl ₃）：δ137.9，137.5，137.4，136.4，128.6，128.6，128.4，128.4，128.2，128.2，127.8，127.7，127.4，83.7，75.4，75.4，74.5，73.8，73.5，73.4，72.8，67.3，63.4，60.6，59.5，27.5，27.4，23.0，22.9。

I-3: productive rate 81%; ¹hNMR(300MHz, D ₂o): δ 4.25(d, 1H, J=7.28Hz), 4.10(dd, 1H, J=4.05Hz, 14.71Hz), 4.04-3.95(m, 1H), 3.92-3.82(m, 2H), 3.79-3.72(m, 2H), 3.48(t, 1H, J=9.46Hz), 3.40-3.24(m, 3H), 3.01(t, 1H, J=12.16Hz), 1.95-1.87(m, 4H), 1.73-1.54(m, 4H); ¹³cNMR(75MHz, D ₂o): δ 76.7,68.0,66.5,64.3,61.8,58.8,55.3,26.9,26.6,22.9,22.8.

Embodiment 4:(2R, 3R, 4R, 5S) preparation of-N, N-(3-oxa--pentamethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-4)

According to the method for embodiment 1 with II-1 and 3-oxa--pentamethylene bromide for raw material prepares compound IV-4 and I-4.

IV-4: productive rate is 67%; ¹hNMR(300MHz, CDCl ₃): δ 7.33-7.14(m, 20H), 4.73-4.64(m, 4H) and, 4.60-4.43(m, 7H), 4.27(dd, 1H, J=5.64Hz, 12.90Hz) and, 4.14(d, 1H, J=6.45Hz), 4.02-3.79(m, 11H); ¹³cNMR(75MHz, CDCl ₃): δ 137.0,136.7,136.5,128.7,128.6,128.4,128.3,128.3,128.2,128.2,128.1,127.6,77.5,77.2,75.0,73.8,73.8,73.5,73.1,72.4,70.5,65.0,61.5,60.5,58.6,54.6.

I-4: productive rate is 87%; ¹hNMR(300MHz, D ₂o): δ 4.44(q, 1H, J=4.59Hz, 13.35Hz), 4.32(d, 1H, J=7.34Hz), 4.21-4.18(m, 2H), 4.14-4.04(m, 2H), 4.00-3.86(m, 5H), 3.55(t, 1H, J=9.4Hz), 3.48-3.38(m, 2H), 3.21(d, 1H, J=5.06Hz), 3.04(t, 1H, J=11.98Hz); ¹³cNMR(75MHz, D ₂o): δ 76.5,76.2,65.3,63.9,60.2,60.1,59.6,56.3,54.7,52.9.

Embodiment 5:(2R, 3R, 4R, 5R) preparation of-N, N-(tetramethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide I-5

According to the method for embodiment 1 with II-2 and Isosorbide-5-Nitrae-dibromobutane for raw material prepares compound IV-5 and I-5

IV-5: productive rate 48%; ¹hNMR(300MHz, CDCl ₃) δ 7.33-7.18(m, 20H), 4.81-4.77(m, 2H), 4.67-4.61(m, 4H), 4.50-4.33(m, 4H), 4.20-4.03(m, 8H), 3.88-3.84(m, 1H), 3.63-3.59(m, 1H) and, 2.17-1.93(m, 4H) ppm; ¹³cNMR(75MHz, CDCl ₃) δ 137.59,137.37,137.19,136.54,128.61128.49,128.30,128.12,127.89,127.65,76.78,74.62,73.49,73.23,71.13,70.07,66.38,64.90,22.10ppm.

I-5: productive rate is 87%; ¹hNMR(400MHz, D2O) δ 4.24(m, 1H), 4.15-4.05(m, 4H) and, 3.92-3.85(m, 1H), 3.80-3.73(m, 2H) and, 3.67-3.55(m, 2H), 3.41-3.34(m, 2H) and, 2.12-2.10(m, 4H) ppm; ¹³cNMR(100MHz, D ₂o) δ 73.21,71.21,67.18,66.38,65.60,61.57,60.60,56.83,22.59,21.79ppm.

Embodiment 6:(2R, 3R, 4R, 5R) preparation of-N, N-(pentamethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-6)

According to the method for embodiment 1 with II-2 and pentamethylene bromide for raw material prepares compound IV-6 and I-6.

IV-6: productive rate 96%; ¹hNMR(300MHz, CDCl ₃) δ 7.30-7.20(m, 20H), 4.92-4.82(m, 2H), 4.68-4.60(m, 5H), 4.50-4.20(m, 7H), 4.11-3.94(m, 4H), 3.59(bs, 1H), 3.44(t, J=11.4Hz, 1H), 2.02-1.77(m, 5H), 1.48-1.45(m, 1H) and ppm; ¹³cNMR(75MHz, CDCl ₃) δ 137.74,137.59,137.29,136.63,128.63,128.57,128.48,128.41,128.26,128.13,127.85,127.79,127.47,74.90,73.99,73.40,73.32,72.12,70.97,63.09,62.47,58.24,55.31,21.08,20.31,20.17ppm.

I-6: productive rate is 90%; ¹hNMR(300MHz, D ₂o) δ 4.29-4.09(m, 5H), 3.92-3.74(m, 2H), 3.70(dd, J=3.6,9.0Hz, 1H), 3.38(t, J=11.7Hz, 1H), 3.24-3.03(m, 3H), 2.06-1.89(m, 2H), 1.77-1.73(m, 3H), 1.56-1.39(m, 1H) and ppm; ¹³cNMR(75MHz, D2O) δ 75.17,71.81,65.54,64.39,63.66,56.90,56.68,55.37,20.68,19.69,19.47ppm.

Embodiment 7:(2R, 3R, 4R, 5R) preparation of-N, N-(hexamethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-7)

Be that raw material prepares compound IV-7 and I-7 according to the method for embodiment 1 with II-2 and 1,6-dibromo-hexane.

IV-7: productive rate 35%; ¹hNMR(300MHz, CDCl ₃) δ 7.34-7.20(m, 20H), 4.99-4.95(m, 1H), 4.85-4.81(m, 1H), 4.74-4.58(m, 5H), 4.51-4.47(m, 1H), 4.42-4.40(m, 1H), 4.33-4.23(m, 3H), 4.17-3.96(m, 5H), 3.85-3.78(m, 2H), 3.72-3.67(m, 1H), 1.87-1.41(m, 8H) ppm; ¹³cNMR(75MHz, CDCl ₃) δ 137.84,137.80,137.22,136.43,128.64128.56,128.44,128.36,128.32,128.29,128.17,127.82,127.77,127.46,80.61,75.47,74.43,73.96,73.57,73.17,72.42,71.95,67.7563.62,60.72,28.05,27.84,23.10,23.00ppm.

I-7: productive rate is 85%; ¹hNMR(300MHz, D ₂o) δ 4.23-4.08(m, 4H), 3.95-3.78(m, 3H), 3.72-3.65(m, 2H) and, 3.31-3.19(m, 3H), 1.87-1.85(m, 4H) and, 1.67-1.55(m, 4H) ppm; ¹³cNMR(75MHz, D ₂o) δ 76.95,71.94,68.40,65.96,64.67,61.39,60.82,55.66,26.79,26.61,23.19,22.65ppm; IR(neat, cm ^-1) 3328,2933,1078.

Embodiment 8:(2R, 3R, 4R, 5R) preparation of-N, N-(3-oxa--pentamethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-8)

According to the method for embodiment 1 with II-2 and 3-oxa--pentamethylene bromide for raw material prepares compound IV-8 and I-8.

IV-8: productive rate 88%; ¹hNMR(400MHz, CDCl ₃) δ 7.24-7.20(m, 20H), 4.98-4.96(m, 1H), 4.90-4.82(m, 2H), 4.75-4.60(m, 5H), 4.51-4.42(m, 3H), 4.36-4.29(m, 4H), 4.19-4.10(m, 4H) and, 4.08-3.76(m, 5H) ppm; ¹³cNMR(75MHz, CDCl ₃) δ 137.66,137.31,136.57,128.54,128.37,128.11,128.00,127.69,74.78,73.35,73.02,72.69,70.01,63.66,60.50,55.56,53.85ppm.

I-8: productive rate is 95%; ¹hNMR(400MHz, D ₂o) δ 4.42(m, 1H), 4.33-4.08(m, 7H), 4.00-3.97(m, 3H) and, 3.82-3.77(m, 2H), 3.47(m, 1H) and, 3.30-3.27(m, 2H) ppm; ¹³cNMR(100MHz, D ₂o) δ 75.07,71.24,64.84,61.15,60.45,60.27,56.82,56.35,55.72ppm; IR(neat, cm ^-1) 3316,1079,1035.

Embodiment 9:(2R, 3S, 4R, 5S) preparation of-N, N-(tetramethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-9)

According to the method for embodiment 1 with II-3 and Isosorbide-5-Nitrae-dibromobutane for raw material prepares compound IV-9 and I-9

IV-9: productive rate 62%; ¹hNMR(300MHz, 350K, DMSO) δ 7.35-7.33(m, 20H), 4.82-4.50(m, 8H) and, 4.35(bs, 1H), 4.13-3.94(m, 7H) and, 3.80-3.77(m, 2H), 3.67-3.56(m, 2H) and, 2.10(bs, 4H) ppm; ¹³cNMR(150MHz, 280K, CDCl ₃): δ 137.01,136.85,136.61,129,14,128.59,128.50,128.22,128.15,128.09,128.04,127.79,73.43,72.98,72.46,72.20,70.08,68.14,65.30,64.76,63.80,52.58,21.61,19.51; ¹³cNMR(150MHz, 243K, CDCl ₃): δ 137.00,136.86,136.77,136.53,128.75,128.71,128.65,128.43,128.31,128.17,127.92,127.51,81.00,75.96,75.01,73.79,73.61,73.28,72.94,72.18,71.98,71.85,71,38,70.03,68.32,67.49,67.16,65.40,65.19,64.60,63.56,62.28,58.97,52.23,24.08,21.77,21.44,19.27ppm.

I-9: productive rate is 98%. ¹HNMR（300MHz，D ₂O）：δ4.26（bs，1H），4.09-3.95（m，4H），3.69-3.52（m，6H），3.07（t，J=11.7Hz，1H），2.13-2.02（m，4H）； ¹³CNMR（75MHz，D ₂O）：δ72.34，71.28，68.09，67.60，63.85，63.37，59.61，56.96，23.52，21.62。

Embodiment 10:(2R, 3S, 4R, 5S) preparation of-N, N-(pentamethylene)-2-benzyloxymethyl-3,4,5-tri-benzyloxy piperidinium bromide (I-10)

According to the method for embodiment 1 with II-3 and pentamethylene bromide for raw material prepares compound IV-10 and I-10

IV-10: productive rate 70%, ¹hNMR(300MHz, 350K, DMSO) δ 7.68-7.08(m, 20H), 4.81-4.41(m, 9H) and, 4.24-3.69(m, 10H), 3.43(bs, 1H) and, 1.94-1.52(m, 6H) ppm, ¹³cNMR(75MHz, 380k, C ₂d ₂cl ₄): δ 138.22,137.78,137.70,129.36,129.26,129.23,129.03,128.98,128.91,128.82,128.76,128.56,76.65,74.59,74.12,73.90,73.22,72.54,70.21,65.31,62.73,61.07,55.24,21.43,21.06,20.86ppm, ¹³cNMR(150MHz, 243K, CDCl ₃): δ 138.20, 137.73, 137.11, 136.96, 136.61, 136.51, 136.45, 130.05, 129.22, 128.92, 128.79, 128.73, 128.69, 128.55, 128.47, 128.39, 128.35, 128.08, 127.95, 127.73, 80.92, 76.44, 75.60, 74.11, 74.03, 73.32, 73.25, 73.13, 72.71, 72.32, 72.24, 71.85, 70.79, 70.21, 68.81, 64.80, 63.76, 62.45, 60.18, 54.56, 54.18, 20.66, 20.44, 20.32, 19.88, 19.05ppm.

I-10: productive rate is 98%; ¹hNMR(300MHz, D ₂o): δ 4.32-4.26(m, 2H), 4.19-4.10(m, 2H), 4.07(dd, J=2.4Hz, 6.0Hz, 1H), 3.95-3.85(m, 2H), 3.67(dd, J=3.6Hz, 9.3Hz, 1H), 3.48-3.44(m, 1H), 3.29-3.22(m, 2H), 2.85(t, J=11.77Hz, 1H), 2.12-1.86(m, 2H), 1.78-1.75(m, 3H), 1.51-1.36(m, 1H); ¹³cNMR(75MHz, D ₂o): δ 74.09,72.46,68.20,63.24,62.52,56.52,56.10,55.36,20.26,19.48,19.09.

Embodiment 11:(2R, 3S, 4R, 5S) preparation of-N, N-(hexamethylene)-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-11)

Be that raw material prepares compound IV-11 and I-11 according to the method for embodiment 1 with II-3 and 1,6-dibromo-hexane.

IV-11: productive rate 57%, ¹hNMR(300MHz, 350K, DMSO) δ 7.59-7.07(m, 20H), 4.84-4.54(m, 8H), 4.40(bs, 1H), 4.25-4.17(m, 2H), 4.07-3.87(m, 6H), 3.79-3.74(m, 1H), 3.62-3.55(m, 2H), 2.01-1.26(m, 8H) and ppm, ¹³cNMR(150MHz, 253K, CDCl ₃): δ 138.25, 137.83, 137.08, 136.99, 136.80, 136.37, 136.33, 128.75, 128.65, 128.52, 128.49, 128.41, 128.34, 128.26, 128.19, 127.99, 127.83, 127.67, 80.81, 76.41, 76.30, 74.09, 73.95, 73.30, 73.20, 72.95, 72.30, 72.22, 72.05, 71.34, 71.20, 70.45, 69.04, 67.43, 66.33, 64.54, 64.46, 63.32, 61.84, 59.86, 53.98, 28.33, 28.07, 27.26, 27.19, 22.99, 22.83, 21.53, 21.17ppm.

I-11: productive rate is 73%; ¹hNMR(300MHz, D ₂o): δ 4.28-4.22(m, 2H), 4.11-4.00(m, 3H), 3.96-3.88(m, 1H), 3.80-3.70(m, 2H), 3.62(dd, J=4.2Hz, 9.9Hz, 1H), 3.48(td, J=2.4Hz, 6.0Hz, 1H), 3.38-3.34(m, 1H), 2.96(t, J=11.7Hz, 1H), 1.89-1.48(m, 8H); ¹³cNMR(75MHz, D ₂o): δ 74.79,72.38,68.29,62.75,62.28,59.66,56.74,27.04,26.46,22.83,22.68.

Embodiment 12:(2R, 3S, 4R, 5S)-N, N-(3-oxa--pentamethylene) preparation of-2-methylol-3,4,5-trihydroxy-piperidinium bromide (I-12)

According to the method for embodiment 1 with II-3 and 3-oxa--pentamethylene bromide for raw material prepares compound IV-12 and I-12.

IV-12: productive rate 72%; ¹hNMR(300MHz, 330K, DMSO) δ 7.34-7.31(m, 20H), 4.77-4.66(m, 6H) and, 4.56-4.40(m, 4H), 4.26-3.68(m, 14H) and ppm; ¹³cNMR(150MHz, 243k, CDCl ₃): δ 138.11,137.68,136.88,136.76,136.62,136.36,129.02,128.91,128.81,128.75,128.73,128.69,128.50,128.37,128.29,128.05,127.73,80.81,76.24,75.42,74.25,73.26,73.19,73.09,72.38,72.17,72.13,71.84,70.95,70.24,68.52,65.19,63.53,61.28,61.07,60.49,60.39,60.22,60.02,59.37,54.69,53.52ppm.

I-12: productive rate is 87%; ¹hNMR(300MHz, D ₂o): δ 4.34-4.27(m, 3H), 4.20-4.05(m, 5H), 3.94-3.81(m, 4H) and, 3.72-3.62(m, 3H), 3.35(m, 1H) and, 3.13-3.10(m, 1H); ¹³cNMR(75MHz, D ₂o): δ 74.48,72.04,67.87,63.16,60.90,60.23,59.89,56.47,54.96.

Embodiment 13:(2S, 3S, 4S) preparation of-N, N-(tetramethylene)-2-methylol-3,4-dihydroxyl Pyrrolidine bromide (1-13)

According to the method for embodiment 1 with II-4 and Isosorbide-5-Nitrae-dibromobutane for raw material prepares compound IV-13 and I-13.

IV-13: productive rate 95%, brown-red oil; ¹hNMR(300MHz, CDCl ₃) δ 7.35-7.24(m, 15H), 4.59-4.46(m, 6H), 4.39-4.24(m, 4H), 4.09-4.04(m, 3H) and, 3.90-3.82(m, 3H), 3.71-3.69(m, 1H) and, 2.19-2.09(m, 4H); ¹³cNMR(75MHz, CDCl ₃) δ 136.8,136.6,136.57,128.78,128.75,128.45,128.42,128.38,128.29,128.1,83.2,80.0,75.0,73.8,72.54,72.51,66.4,65.5,59.6,22.1,21.6.

I-13:35mg, productive rate 95%; ¹hNMR(300MHz, D ₂o) δ 4.41-4.37(m, 1H), 4.21(dd, J=6.9,3.6Hz, 1H), 4.04-4.02(m, 2H) and, 3.81-3.49(m, 7H), 2.19-2.06(m, 4H); ¹³cNMR(75MHz, D ₂o) δ 77.0,76.8,73.6,67.3,66.6,59.2,57.1,21.2,20.4.

Embodiment 14:(2S, 3S, 4S) preparation of-N, N-(pentamethylene)-2-methylol-3,4-dihydroxyl Pyrrolidine bromide (I-14)

According to the method for embodiment 1 with II-4 and Isosorbide-5-Nitrae-dibromobutane for raw material prepares compound IV-14 and I-14.

IV-14: yield 92%, red solid; ¹hNMR(300MHz, CDCl ₃) δ 7.32-7.29(m, 15H), 4.62-4.46(m, 6H), 4.34-4.25(m, 4H), 4.18-4.03(m, 2H) and, 3.88-3.82(m, 3H), 3.62-3.57(m, 2H) and, 2.03-1.71(m, 6H); ¹³cNMR(75MHz, CDCl ₃) δ 136.7,136.5,128.6,128.5,128.3,128.2,128.1,128.0,82.7,79.6,77.4,73.5,72.4,64.7,62.9,62.7,56.7,21.5,21.0,20.8.

I-14:39mg, productive rate 96%; ¹hNMR(300MHz, D ₂o) δ 4.31-4.30(m, 1H), 4.16-4.14(m, 3H), 3.99(dd, J=12.9,2.4Hz, 1H) and, 3.89-3.56(m, 5H), 3.48-3.44(m, 1H) and, 2.08-1.84(m, 5H), 1.59-1.55(m, 1H); ¹³cNMR(75MHz, D ₂o) δ 80.5,75.9,73.3,64.4,63.4,56.8,56.4,21.2,20.6,20.0.

Embodiment 15:(2R, 3R, 4R)-N, N-(hexamethylene)-2-methylol-3,4-dihydroxyl Pyrrolidine bromide I-15:

Be that raw material prepares compound IV-15 and I-15 according to the method for embodiment 1 with II-4 and 1,6-dibromo-hexane.

IV-15: yield 94%; (2S, 3S, 4S)-N, N-(hexamethylene)-2-benzyloxymethyl-3,4-benzyloxy Pyrrolidine bromide IV-15: faint yellow solid; ¹hNMR(300MHz, CDCl ₃) δ 7.34-7.26(m, 15H), 4.65-4.44(m, 6H), 4.35-4.27(m, 3H), 4.20-3.98(m, 5H) and, 3.85-3.76(m, 2H), 3.55-3.50(m, 1H) and, 2.03-1.73(m, 8H); ¹³cNMR(75MHz, CDCl ₃) δ 136.7,136.5,136.3,128.6,128.57,128.3,128.2,128.1,82.9,79.4,78.7,73.6,72.4,67.6,66.6,64.9,61.0,27.3,27.0,23.1,23.0.

I-15: yield 98%; ¹hNMR(300MHz, D ₂o) δ 4.38(bs, 1H): 4.19(dd, J=7.8,2.7Hz, 1H), 4.09(m, 2H) and, 3.86-3.68(m, 4H), 3.59-3.53(m, 1H) and, 3.48-3.42(m, 2H), 1.90-1.59(m, 8H); ¹³cNMR(75MHz, D ₂o) δ 81.9,76.9,73.6,68.3,67.8,60.8,56.8,26.8,26.3,22.7,22.4.

Embodiment 16:(2S, 3S, 4S) preparation of-N, N-(3-oxa--pentamethylene)-2-methylol-3,4-dihydroxyl Pyrrolidine bromide (IV-16)

Be that raw material prepares compound IV-16 and I-16 according to the method for embodiment 1 with the bromo-pentane of II-4 and 3-oxa--1,5-bis-.

IV-16: productive rate 96%; ¹hNMR(300MHz, CDCl ₃) δ 7.37-7.24(m, 15H), 4.70-4.40(m, 10H), 4.23-4.08(m, 3H), 4.05-3.85(m, 6H) and, 3.73-3.65(m, 1H), 3.50-3.46(m, 1H); ¹³cNMR(75MHz, CDCl ₃) δ 136.6,136.4,136.2,128.7,128.6,128.4,128.35,128.31,128.27,128.0,82.2,79.3,77.6,73.7,72.5,72.4,64.4,63.4,62.1,61.2,61.0,55.7.

I-16: yield 91%; ¹hNMR(300MHz, D ₂o) δ 4.43-4.42(m, 1H), 4.24(dd, J=7.5,3.9Hz, 1H), 4.15-4.13(m, 2H), 4.06-3.95(m, 6H), 3.91-3.78(m, 2H), 3.72-3.66(m, 1H) and, 3.56(dd, J=10.8,2.1Hz, 1H), 3.42(dd, J=12.0,2.1Hz, 1H); ¹³cNMR(75MHz, D ₂o) δ 81.5,75.6,73.3,63.8,62.6,61.9,61.1,56.5,56.1.

Embodiment 17:(2R, 3R, 4R) preparation of-N, N-(tetramethylene)-2-methylol-3,4-dihydroxyl Pyrrolidine bromide (I-17)

According to the method for embodiment 1 with II-5 and Isosorbide-5-Nitrae-dibromobutane for raw material prepares compound IV-17 and I-17.

IV-17:212mg, productive rate 90%, pale yellow oil; =+3.6(c0.55, CH ₃oH); ¹hNMR(300MHz, CDCl ₃) δ 7.33-7.24(m, 15H), 4.59-4.45(m, 6H), 4.37-4.23(m, 4H), 4.10-3.99(m, 3H) and, 3.90-3.79(m, 3H), 3.73-3.67(m, 1H) and, 2.24-2.16(m, 4H); ¹³cNMR(75MHz, CDCl ₃) δ 136.7,136.5,136.4,128.61,128.58,128.3,128.2,128.1,128.0,83.1,79.9,77.4,74.8,72.3,66.3,65.4,59.4,22.0,21.4.

I-17: orange, 35mg, productive rate 95%; ¹hNMR(300MHz, D ₂o) δ 4.39-4.37(m, 1H), 4.20(dd, J=6.9,3.6Hz, 1H), 4.02-4.01(m, 2H) and, 3.80-3.48(m, 7H), 2.15-2.04(m, 4H); ¹³cNMR(75MHz, D ₂o) δ 77.0,76.8,73.5,67.3,66.6,59.2,57.1,21.2,20.4.

Embodiment 18:(2R, 3R, 4R) preparation of-N, N-(pentamethylene)-2-methylol-3,4-dihydroxyl Pyrrolidine bromide (I-18)

According to the method for embodiment 1 with II-5 and pentamethylene bromide for raw material prepares compound IV-18 and I-18.

IV-18 is 216mg altogether, yield 89%, white solid, M.p.104-111 DEG C; ¹hNMR(300MHz, CDCl ₃) δ 7.34-7.24(m, 15H), 4.61-4.49(m, 6H), 4.45-4.40(m, 1H), 4.37-4.30(m, 2H), 4.24(d, J=5.7Hz, 1H), 4.15(dd, J=12.9,2.4Hz1H), 4.04-3.91(m, 3H), 3.82(dd, J=12.6,5.4Hz, 1H), 3.70-3.52(m, 2H), 2.02-1.85(m, 5H), 1.62(bs, 1H); ¹³cNMR(75MHz, CDCl ₃): 136.6,136.54,136.51,128.6,128.5,128.2,128.19,128.15,128.10,128.0,82.7,79.6,77.3,73.4,72.3,64.7,63.0,62.7,56.7,21.5,21.0,20.8.

I-18: light red solid, 39mg, productive rate 96%; ¹hNMR(300MHz, CDCl ₃) δ 4.33-4.29(m, 1H), 4.15(dd, J=3.6,8.1Hz, 1H), 4.06-3.83(m, 2H), 3.81-3.67(m, 2H), 3.59(dt, J=4.2,14.1Hz, 1H), 3.52-3.34(m, 3H), 3.30-3.25(m, 1H), 1.95-1.68(m, 5H), 1.47-1.33(m, 1H) ¹³cNMR(75MHz, CDCl ₃) δ 80.5,75.9,73.3,64.4,63.4,56.8,56.4,21.2,20.6,20.0.

Embodiment 19:(2S, 3S, 4S) preparation of-N, N-(hexamethylene)-2-methylol-3,4-dihydroxyl Pyrrolidine bromide (I-19)

Be that raw material prepares compound IV-19 and I-19 according to the method for embodiment 1 with II-5 and 1,6-dibromo-hexane.

IV-19:247mg, yield 92%.Pale yellow oil; ¹hNMR(300MHz, CDCl ₃) δ 7.32-7.22(m, 15H), 4.66-4.44(m, 6H), 4.37-4.30(m, 2H), 4.26-4.12(m, 3H), 4.08-3.94(m, 3H), 3.86-3.74(m, 2H), 3.57-3.50(m, 1H), 2.08-2.03(m, 2H), 1.86-1.77(s, 2H), 1.72-1.59(m, 4H); ¹³cNMR(75MHz, CDCl ₃) δ 136.7,136.5,136.3,128.6,128.5,128.3,128.29,128.25,128.1,82.9,79.4,78.7,73.6,72.4,67.6,66.6,64.9,61.0,27.3,27.0,23.1,23.0.

I-19: yellow oil, 37mg, productive rate 95%; ¹hNMR(300MHz, D ₂o) δ 4.39(m, 1H), 4.20(dd, J=7.8,2.4Hz, 1H), 4.11-4.09(m, 2H) and, 3.87-3.68(m, 4H), 3.60-3.54(m, 1H) and, 3.51-3.41(m, 2H), 1.91-1.59(m, 8H); ¹³cNMR(75MHz, D ₂o) δ 81.9,76.9,73.7,68.2,67.8,60.8,56.8,26.8,26.3,22.7,22.4.

Embodiment 20:(2R, 3R, 4R) preparation of-N, N-(3-oxa--pentamethylene)-2-methylol-3,4-dihydroxyl Pyrrolidine bromide (I-20)

Be that raw material prepares compound IV-20 and I-20 according to the method for embodiment 1 with the bromo-pentane of II-5 and 3-oxa--1,5-bis-.

IV-20:235mg, productive rate 92%, colorless oil; ¹hNMR(300MHz, CDCl ₃): δ 7.29-7.16(m, 15H), 4.68(dd, J=12.3,5.7Hz, 1H) and, 4.52-4.25(m, 10H), 4.14-4.04(m, 2H) and, 3.94-3.74(m, 6H), 3.59(dt, J=12.6,3.6Hz, 1H) and, 3.38-3.34(m, 1H); ¹³cNMR(75MHz, CDCl ₃): 136.6,136.4,136.2,128.7,128.6,128.35,128.30,128.27,128.0,82.2,79.3,77.7,73.7,72.5,72.4,64.5,63.4,62.1,61.2,61.0,55.7.

I-20: coffee-like oily matter, 37mg, yield 91%; ¹hNMR(300MHz, D ₂o) δ 4.40(m, 1H), 4.22(dd, J=7.2,3.6Hz, 1H), 4.14-4.12(m, 2H), 4.05-3.86(m, 7H), 3.84-3.75(m, 1H), 3.71-3.65(m, 1H) and, 3.55(dd, J=12.9,2.1Hz, 1H), 3.42-3.38(dd, J=11.4,1.8Hz, 1H); ¹³cNMR(75MHz, D ₂o) δ 81.5,75.6,73.3,63.8,62.6,61.9,61.1,56.5,56.0.

Embodiment 21:(2S, 3S, 4S, 5S)-N, N-(tetramethylene)-2,5-preparation of dihydroxymethyl-3,4-dihydroxyl-Pyrrolidine bromide (I-21)

According to the method for embodiment 1 with II-6 and Isosorbide-5-Nitrae-dibromobutane for raw material prepares compound IV-21 and I-21.

IV-21:170mg, productive rate 88%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 7.34-7.25(m, 15H), 4.69-4.47(m, 10H) and, 4.26-4.10(m, 6H), 3.25(s, 3H) and, 2.39-2.28(m, 4H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 136.6,128.6,128.5,128.4,128.4,128.3,128.0,96.6,82.0,81.9,74.5,74.4,73.5,72.5,72.4,65.8,63.7,59.8,59.7,56.0,21.8.

I-21:62mg, productive rate 100%; ¹hNMR(300MHz, D ₂o): δ (ppm) 4.28-4.24(m, 2H), 4.05-3.94(m, 4H), 3.74-3.69(m, 4H) and, 3.64-3.56(m, 2H), 2.17-2.01(m, 4H); ¹³cNMR(75MHz, D ₂o): δ (ppm) 76.4,75.3,59.5,57.2,20.8.

Embodiment 22:(2S, 3S, 4S, 5S)-N, N-(pentamethylene)-2,5-preparation of dihydroxymethyl-3,4-dihydroxyl-Pyrrolidine bromide (I-22)

According to the method for embodiment 1 with II-6 and 1,5-dibromo penta for raw material prepares compound IV-22 and I-22.

IV-22:130mg, productive rate 68%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 7.35-7.26(m, 15H), 4.78-4.48(m, 10H) and, 4.20-3.97(m, 6H), 3.75-3.63(m, 2H) and, 3.24(s, 3H), 2.17-2.14(m, 4H) and, 1.76-1.75(m, 2H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 136.6,136.5,128.5,128.3,128.2,128.0,96.6,82.1,82.1,77.6,77.3,77.1,76.7,73.6,72.4,72.3,57.6,56.0,21.3.

I-22:68mg, productive rate 100%; ¹hNMR(300MHz, D ₂o): δ (ppm) 4.36-4.34(m, 2H), 4.10-4.00(m, 4H), 3.99-3.96(m, 2H) and, 3.51-3.42(m, 2H), 1.88-1.81(m, 4H) and, 1.65-1.57(m, 2H); ¹³cNMR(75MHz, D ₂o): δ (ppm) 77.1,76.9,57.4,57.2,20.9,20.6.

Embodiment 23:(2S, 3S, 4S, 5S)-N, N-(hexamethylene)-2,5-preparation of dihydroxymethyl-3,4-dihydroxyl-Pyrrolidine bromide (I-23)

Be that raw material prepares compound IV-23 and I-23 according to the method for embodiment 1 with II-6 and 1,6-dibromo-hexane

IV-23:120mg, productive rate 51%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 7.29-7.16(m, 15H), 4.80-4.75(m, 2H) and, 4.66-4.37(m, 8H), 4.10-4.00(m, 4H) and, 3.89-3.69(m, 6H), 3.18(s, 3H) and, 2.10-1.93(m, 4H), 1.76-1.58(m, 4H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 136.5,136.3,128.6,128.6,128.5,128.5,128.3,128.0,96.7,82.1,82.1,77.9,77.6,77.3,77.1,76.7,73.7,72.4,72.3,66.6,64.3,61.6,56.1,28.3,23.2.

I-23:37mg, productive rate 100%; ¹hNMR(300MHz, D ₂o): δ (ppm) 4.22-4.20(m, 2H), 4.05-3.93(m, 4H), 3.79-3.71(m, 4H) and, 3.57-3.49(m, 2H), 3.21(s, 1H) and, 1.89-1.81(m, 2H), 1.80-1.68(m, 4H) and, 1.47-1.43(m, 2H); ¹³cNMR(75MHz, D ₂o): δ (ppm) 80.7,76.2,61.8,57.6,27.7,22.8.

Embodiment 24:(3R, 4R, 5R, 6R)-N, N-(tetramethylene)-3,4,5,6-preparation of tetrahydroxy azepan bromide (I-24)

According to the method for embodiment 1, with II-7 and Isosorbide-5-Nitrae-dibromobutane for raw material prepares compound IV-24 and I-24.

IV-24:312mg, productive rate 86%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 7.34-7.21(m, 20H), 4.72-4.54(m, 8H) and, 4.00-3.90(m, 2H), 3.84-3.80(m, 8H) and, 3.59-3.55(m, 2H), 2.14-2.12(m, 4H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 137.6,137.0,128.5,128.4,128.2,128.1,127.8,78.7,73.5,72.8,71.7,67.5,59.5,21.1.

I-24:68mg, productive rate 98%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 4.29-4.27(m, 2H), 3.85(m, 2H) and, 3.78-3.66(m, 6H), 3.35-3.30(m, 2H) and, 2.27(m, 4H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 73.4,68.3,65.7,60.5,21.0.

Embodiment 25:(3R, 4R, 5R, 6R)-N, N-(pentamethylene)-3,4,5,6-preparation of tetrahydroxy azepan bromide (I-25)

According to the method for embodiment 1, with II-7 and pentamethylene bromide for raw material prepares compound IV-25 and I-25.

IV-25:360mg, productive rate 91%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 7.37-7.19(m, 22H), 4.68-4.49(m, 8H) and, 3.92-3.88(m, 2H), 3.82-3.73(m, 8H) and, 3.50-3.42(m, 2H), 1.86-1.70(m, 9H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 137.4,136.7,128.6,128.5,128.3,128.1,128.0,76.1,73.6,69.5,62.6,58.0,20.4,20.0.

I-25:107mg, productive rate 96%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 4.26-4.23(m, 2H), 3.91(m, 2H) and, 3.73-3.65(m, 2H), 3.60-3.43(m, 4H) and, 3.34-3.29(m, 2H), 1.97-1.86(m, 4H) and, 1.68-1.60(m, 2H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 72.9,64.0,63.2,59.1,20.4,19.7.

Embodiment 26:(3R, 4R, 5R, 6S)-N, N-(tetramethylene)-3,4,5,6-preparation of tetrahydroxy azepan bromide (I-26)

According to the method for embodiment 1, with II-8 and Isosorbide-5-Nitrae-dibromobutane for raw material prepares compound IV-26 and I-26.

IV-26:329mg, productive rate 86%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 7.35-7.15(m, 34H), 4.71-4.41(m, 8H) and, 3.92-3.88(m, 2H), 4.37-4.24(m, 2H), 4.11-4.03(m, 2H), 3.95(m, 1H), 3.87-3.85(m, 2H), 3.80-3.77(m, 1H), 3.74-3.65(m, 2H), 3.33-3.23(m, 2H), 2.30-2.17(m, 2H), 2.02-2.00(m, 1H), 1.88-1.85(m, 1H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 137.0,136.9,128.5,128.5,128.4,128.4,128.2,128.1,128.0,128.0,127.8,78.9,77.3,75.9,75.4,73.6,73.2,72.7,72.2,71.8,68.3,64.6,59.5,21.5,20.6.

I-26:89mg, productive rate 98%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 4.29-4.27(m, 1H), 3.98-3.92(t, J=9.0Hz, 1H) and, 3.80-3.58(m, 8H), 3.46-3.35(m, 2H) and, 2.20-2.18(m, 4H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 75.3,75.1,69.4,67.1,66.3,62.5,61.1,21.4,20.5.

Embodiment 27:(3R, 4R, 5R, 6S)-N, N-(pentamethylene)-3,4,5,6-preparation of tetrahydroxy azepan bromide (I-27)

According to the method for embodiment 1, with II-8 and pentamethylene bromide for raw material prepares compound IV-27 and I-27.

IV-27:331mg, productive rate 86%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 7.36-7.12(m, 33H), 4.70-4.60(m, 4H), 4.56-4.45(m, 2H), 4.41-4.30(m, 2H), 4.27-4.19(m, 2H), 4.08-4.04(m, 1H), 3.94-3.75(m, 7H), 3.56-3.52(m, 1H), 3.26(m, 1H), 1.77(m, 4H), 1.26(m, 2H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 137.6,137.0,136.9,136.8,128.6,128.5,128.4,128.4,128.3,128.2,128.2,128.1,128.0,128.0,127.9,78.4,75.3,74.9,73.6,73.0,72.7,71.9,70.2,64.9,60.6,59.1,58.1,20.4,20.3,20.2.

I-27:117mg, productive rate 99%; ¹hNMR(300MHz, CDCl ₃): δ (ppm) 4.31-4.28(m, 1H), 4.04-3.93(m, 2H) and, 3.84-3.76(m, 1H), 3.73-3.58(m, 5H), 3.53-3.48(m, 1H), 3.44-3.26(m, 1H), 3.18-3.13(m, 1H), 2.01-1.78(m, 4H), 1.77-1.55(m, 2H); ¹³cNMR(75MHz, CDCl ₃): δ (ppm) 75.1,75.0,67.3,66.0,65.0,63.4,62.2,58.9,20.5,19.7,19.7.

The structural formula of the Azasugar compound obtained by above-described embodiment is as follows:

Formula I-22 formula I-23 formula I-24 formula I-25 formula I-26 formula I-27.

Embodiment 28: spiro-azasugar quaternary ammonium salt compound of the present invention is tested Glycosylase inhibition

1) test materials and source

The spiro-azasugar quaternary ammonium salt compound that test compound: embodiment 1-27 obtains.

Test materials: all 4-nitrophenol pyranoside matrix, disaccharides and Glycosylase (comprising alpha-glucosidase, beta-glucosidase, alpha-galactosidase, beta-galactosidase enzymes, alpha-Mannosidase, beta-Mannosidase, α, α-trehalase and amyloglucosidase) are all purchased from Sigma-Aldrich.

2) test method

Dynamics research carries out in the 50mM Trisodium Citrate/phosphoric acid buffer of 37 DEG C.According to the difference of matrix, the enzyme concn of preparation is 0.1-0.5mg/mL.Active testing, is tested under the optimum activity pH of often kind of enzyme for matrix with 4-nitrophenol pyranoside.Matrix, enzyme solution and inhibitor (volution azasugar quaternary ammonium salt) are cultivated 30 minutes at 37 DEG C, then in ultraviolet-visible pectrophotometer, starts reaction, measure its absorption to 400nm wavelength light.Finally use GraFit program to carry out data analysis and [specifically refer to Leatherbarrow, R.J.Grafit4.0; ErithacusSoftware:Staines, UK, 1998.].

3) evaluation result

Spiro-azasugar quaternary ammonium salt compound provided by the invention to the inhibit activities result of Glycosylase as shown in table 1-table 5: table 1

(): inhibiting rate during 1000 μMs of concentration

Table 2

(): inhibiting rate during 1000 μMs of concentration

Table 3

(): inhibiting rate during 1000 μMs of concentration

Table 4

(): inhibiting rate during 1000 μMs of concentration

Table 5

(): inhibiting rate during 1000 μMs of concentration

Above bioassay result shows, spiro-azasugar quaternary ammonium salt compound of the present invention shows good inhibit activities to above-mentioned various Glycosylase.

More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.It should be noted that in addition, each concrete technical characteristic described in above-mentioned embodiment, in reconcilable situation, can be combined by any suitable mode, in order to avoid unnecessary repetition, the present invention illustrates no longer separately to various possible array mode.

In addition, also can carry out arbitrary combination between various different embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.

Claims

1. a spiro-azasugar quaternary ammonium salt compound, is characterized in that, the compound that described volution azasugar quaternary ammonium compound is structure shown in formula Ia;

In formula Ia, X is fluorine, chlorine, bromine or iodine; N is the integer of 1-3; Two m are identical or different, be separately 0,1 or 2, Y be CH ₂or O;

Wherein, when n is 1, R ³and R ⁴being asynchronously hydrogen, is hydrogen or methylol independently of one another; When n is 2 or 3, R ³and R ⁴be hydrogen or methylol independently of one another.

2. spiro-azasugar quaternary ammonium salt compound according to claim 1, wherein, the spiro-azasugar quaternary ammonium salt compound of structure shown in described formula Ia is any one in following formula I-1 ~ formula I-27;

3. a preparation method for spiro-azasugar quaternary ammonium salt compound, is characterized in that, the method comprises the steps:

Wherein, R ¹and R ²independently of one another for phenyl, benzyl, hydrogen atom on phenyl ring by hydroxyl, carbonatoms be the alkoxyl group of 1-4, any one in the ether of at least one in nitro and the halogen benzyl, the carbonatoms that replace to be the acyl group of 2-10 and carbonatoms be 2-10, benzal and cyclohexylidene; X is fluorine, chlorine, bromine or iodine; Two m are identical or different, be separately 0,1 or 2, Y be CH ₂or O; Wherein, when n is 1, R ³and R ⁴being asynchronously hydrogen, is hydrogen or methylol independently of one another; When n is 2 or 3, R ³and R ⁴be hydrogen or methylol independently of one another.

4. spiro-azasugar quaternary ammonium salt compound described in claim 1 or 2 preparing glycosidase inhibitor, prevent and/or treat the medicine of diabetes, prevent and/or treat the medicine of gaucher's disease, the application prevented and/or treated in the medicine of tumour and antiviral.

5. application according to claim 4, wherein, described Glycosylase is at least one in alpha-glucosidase, beta-glucosidase, alpha-galactosidase, beta-galactosidase enzymes, alpha-Mannosidase, beta-Mannosidase, alpha-L-fucosidase, beta-glucuronidase, α, α-trehalase and alpha-L-Rhamnosidase.

6. a medicine, is characterized in that, the activeconstituents of this medicine contains spiro-azasugar quaternary ammonium salt compound described in claim 1 or 2.

7. medicine according to claim 6, wherein, described medicine be glycosidase inhibitor, prevent and/or treat the medicine of diabetes, prevent and/or treat the medicine of gaucher's disease, prevent and/or treat in the medicine of tumour and antiviral any one.

8. medicine according to claim 7, wherein, described Glycosylase is at least one in alpha-glucosidase, beta-glucosidase, alpha-galactosidase, beta-galactosidase enzymes, alpha-Mannosidase, beta-Mannosidase, alpha-L-fucosidase, beta-glucuronidase, α, α-trehalase and alpha-L-Rhamnosidase.