CN106467563B

CN106467563B - The synthetic method and its midbody compound of chondroitin sulfate tetrose

Info

Publication number: CN106467563B
Application number: CN201610696173.0A
Authority: CN
Inventors: 李中军; 姚望; 沙勐; 张肖; 孟祥豹; 李树春; 李庆
Original assignee: Peking University
Current assignee: Peking University
Priority date: 2015-08-21
Filing date: 2016-08-19
Publication date: 2019-08-06
Anticipated expiration: 2036-08-19
Also published as: CN106467563A

Abstract

This application discloses a kind of synthetic methods of chondroitin sulfate tetrose.Using hyaluronate as starting material; successively through digesting; protecting group protection; protecting group conversion; selectivity deprotection and hydroxyl oxidation and selective reduction obtain the midbody compound as shown in formula O; midbody compound shown in recycling formula O synthesizes chondroitin sulfate tetrose, and being specifically defined for substituent group sees specification in formula O.In addition, additionally providing the midbody compound of synthesis chondroitin sulfate tetrose.The synthetic route of the relatively existing document of synthetic method provided herein is easy, is not necessarily to glycosylation, reaction yield is high, has a good application prospect.

Description

The synthetic method and its midbody compound of chondroitin sulfate tetrose

Technical field

The application belongs to technical field of chemistry, synthetic method more specifically to chondroitin sulfate tetrose and wherein Intermediate compounds therefor.

Background technique

Chondroitin sulfate is a kind of glycosaminoglycan being widely present in vivo, and structure is by glucuronic acid and N- acetylamino Galactolipin is constituted, and different degrees of sulphation, thus referred to as chondroitin sulfate may be present in hydroxyl in its structure.Chondroitin sulfate Element forms proteoglycans with the albumen covalent bond in extracellular matrix, is distributed widely in space between cells, is a kind of particularly significant Glycosaminoglycan.Chondroitin sulfate takes part in many particularly important biochemical processes in vivo, such as: take part in cell-ECM phase Interaction inhibits the regeneration of neural axon, influences the diffusion and transfer of cancer cell, in conjunction with extracellular numerous cell factors Deng.

In recent years, it has been found that the different subtype of chondroitin sulfate has different sulphation modifications that is, in specific hydroxyl Chondroitin sulfate A (CSA), C, E can mediate different physiologic functions.For example, chondroitin sulfate E can conspicuousness inhibition Dorsal ganglion The growth of ganglion cell's aixs cylinder, and chondroitin sulfate A (CSA) and C are then without this effect.To study different sulfation sites and sulphation number Influence to chondroitin sulfate biological function, need to solve these chondroitin sulfates with specific sulphation pattern first can Acquired problem.Traditional extracts by extracting from natural origin, such as from the cartilage of animal, can only obtain different type sulphur The mixture of aching and limp ossein further isolates and purifies and is difficult to carry out.These extracts can satisfy a degree of drug It learns research to need, such as can be used for the treatment etc. of osteoarthropathy.However, the mixture with different sulfation sites can not expire The deeper medicineization research of foot.Therefore, chemical synthesis is that current obtain has clear sulfation sites, and structure it is uniform can Carry out the only effective method of the chondroitin sulfate of deeper biology and medicineization research.

In the prior art, chondroitin sulfate is synthesized usually with glucuronic acid and N- acetylamino gala by chemical means Sugar or N-acetylglucosamine monosaccharide are raw material, carry out oligosaccharides by conventional protecting group protection also glycosylation operation Assemble (Jean-Claude Jacquinet et al, From Polymer to Size-Defined Oligomers:A Highly Divergent and Stereocontrolled Construction of Chondroitin Sulfate A, C,D,E,K,L,and M Oligomers from a Single Precursor:Part 2.Chem.Eur.J.2009,15, 9579-9595.).Such strategy synthesis step is more, and route is long, operates and cause the receipts of whole route due to being related to glycosylation Rate is relatively low.Currently, only synthesis chondroitin sulfate tetrose just needs about 23 steps to react, yield is less than 1%.Therefore, new sulphur is developed The chemical synthesis process of aching and limp ossein oligosaccharides, for promoting the medicineization research of chondroitin sulfate to be of great significance.

Currently, being both needed to be related to the exposed hydroxyl on the oligosaccharide backbone having had been built up in the post-synthesis phase of chondroitin sulfate Base carries out the deprotection of sulfonated and last full sugar.Therefore, how rapid build can carry out among the oligosaccharides of sulphation modification Body is a problem to be solved.An object of the application is provided and such a can directly be carried out in the oligosaccharides of subsequent sulphuric acid modification The synthetic method of mesosome.

Summary of the invention

Present inventor using it is more easy to produce, be easier to extract obtained Sodium Hyaluronate as raw material, by raw material into Row hydrolysis has been chemically modified to obtain and can be used for chondroitin sulfate A (CSA), and the key intermediate of tri- kinds of tetroses of C, E synthesis overcomes existing Deficiency existing for technology.

First purpose of the application is to provide a kind of synthetic method of chondroitin sulfate tetrose.

Second purpose of the application is to provide the midbody compound for above-mentioned synthetic method.

In the embodiment of the application, a kind of synthetic method of chondroitin sulfate tetrose is provided, the method includes The synthesis of chondroitin sulfate tetrose key intermediate, includes the following steps:

(1) formula A compound (i.e. hyaluronic acid M salt) is in the weak acid buffer solution containing sodium chloride, in hyaluronic acid Formula B compound is obtained under the action of enzyme；Here, the hyaluronidase derives from animal testis, vigor 400-1000IU/ mg；Preferably, bull testis or sheep testicle are selected from；More preferably bull testis；

(2) alcohol R of the formula B compound in inorganic acid₁Esterification is carried out in OH solution, and the carboxyl of formula B compound is carried out Esterification；Then utilize acid anhydrides R₂-O-R₂, to formula B compound, remaining all hydroxyl carries out acylated protection under alkaline condition, obtains To formula C compound；

Here, acid anhydrides R₂-O-R₂In, R₂Selected from aliphatic acyl radical, unsubstituted benzoyl or substituted benzoyl；

(3) formula C compound in the presence of a base, selectively removing reducing end N-acetylglucosamine in organic solvent 1 hydroxyl obtains formula D compound；

(4) formula D compound reacts under base catalysis with three haloacetonitriles, obtains the formula E compound of end group ester containing imines；

(5) formula E compound obtains formula F compound under acid catalysis

(6) formula F compound in presence of an acid, in organic solvent, with alcohol R₄-R₃OH reaction, obtains formula G compound；

Here, alcohol R₄-R₃R in OH₃Selected from-CH₂CH₂-(OCH₂CH₂) n-, n 1-100, aliphatic alkane subunit, fat Race's alkene subunit, or-CH₂-(OCH₂CH₂) m-, m is 1-100 here；Preferably, R₃For-CH₂-O-CH₂CH₂-；R₄Selected from nitrine Base, alkynyl, biotin, substituted amino, aldehyde radical, thioether, the substituted amino refers to the amino protected by protecting group, described Protecting group is selected from benzyloxy carbonyl acyl group, tertiary butyloxycarbonyl acyl group, trichloroacetyl, trifluoroacetyl group；Preferably, R₄-R₃OH is N₃- CH₂-OCH₂CH₂-OH；

(7) formula G compound is in acid or the alcohol R of alkali₁In OH solution, R all on formula G compound is sloughed₂, obtain formula H chemical combination Object；

(8) formula H compound is in R₅CHO or R₅CHO contracts in the presence of two fatty alcohols, the selective protection formula Hization under acid catalysis 4,6 dihydroxy and 4 ' of object are closed, 6 ' dihydroxy obtain compound of formula I；

Here, R₅CHO or R₅Two fatty alcohols of CHO contracting (such as R₅CH(OCH₃)₂) in R₅Selected from phenyl or substituted phenyl；

(9) compound of formula I is in carboxylic acid halides R₇X or acid anhydrides R₇-O-R₇Under the action of, it is acylated under alkaline condition, obtains formula J Compound；

Here, carboxylic acid halides R₇X or acid anhydrides R₇-O-R₇In R₇Selected from C1-C4 aliphatic acyl radical, unsubstituted benzoyl or Substituted benzoyl；Carboxylic acid halides R₇X in X is selected from chlorine, bromine or iodine, it is preferable that is selected from chlorine or bromine；

(10) formula J compound is under the existence condition of acid and reducing agent, exposed 4 hydroxyls of selectivity, so that 6 are R₅- CH₂Protection, obtains formula K compound；

(11) formula K compound aoxidizes 4,4 ' position hydroxyls under oxidant existence condition, obtains formula L compound；

(12) formula L compound is under reducing agent existence condition, selective two ketone by 4 and 4 ' positions in formula L compound Carbonyl reduction is alcohol, and two hydroxyl configurations of the configuration of two newly-generated hydroxyls and step (11) Chinese style K are on the contrary, obtain The formula M compound of galactose configuration；

(13) formula M compound removes two benzyls or substituted benzyl of 6 and 6 ' positions under oxidant or reducing agent effect Base obtains tetrol, i.e. formula O compound；

Alternatively, following step (10a)-(14) in step (10)-(13) are replaced:

(10a) formula J compound hydrolyzes two benzals or substituted benzal, obtains formula K ' under the existence condition of acid Compound；

(11a) formula K ' compound is in chlorosilane R₈R₈R₉In the presence of SiCl and alkali, regioselectivity to two N- acetyl The 6 of Glucosamine, 6 ' position hydroxyls carry out silanization, obtain formula L ' compound；

(12a) formula L ' compound is under oxidant existence condition, selectively by two N- acetyl in formula L ' compound The hydroxyl of the 4 of Glucosamine and 4 ' positions is oxidized to ketone carbonyl, obtains formula M ' compound；

(13a) formula M ' compound is under reducing agent existence condition by the ketone carbonyl of the 4 of N-acetylglucosamine and 4 ' positions Stereoselectivity is reduced to axial bond hydroxyl, obtains formula N ' compound；

(14) formula N ' compound is in the presence of acid or fluorine ion reagent 6 and 6 ' in removing N- acetylgalactosamine residues The silicon ether of position, obtains tetrol, i.e. formula O compound；

(15) formula O compound synthesis chondroitin sulfate tetrose is utilized；

Here, in formula A- formula O compound, Ac is acetyl group；

Formula C- formula O compound and alcohol R₁In OH, R₁Selected from unsubstituted benzyl or substituted benzyl, C1-C4 alkyl or Allyl；Here, the substituted benzyl refers to phenyl ring replaced one or more substituent group, and the substituent group is selected from Halogen (such as fluorine, chlorine, bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl Base or tert-butyl), C1-C4 halogenated alkyl (such as trifluoromethyl), C1-C4 alkoxy (such as methoxyl group, ethyoxyl, positive third oxygen Base, isopropoxy, n-butoxy, isobutoxy or tert-butoxy)；Also, the substituent group is in any position of phenyl ring；It is preferred that Ground, R₁Selected from methyl, ethyl, allyl, benzyl, 4- methoxy-benzyl is more preferably selected from methyl, ethyl, benzyl；

In formula C- formula G compound, formula J- formula O compound, R₂And R₇It is each independently selected from aliphatic acyl radical, unsubstituted Benzoyl or substituted benzoyl；Here, the aliphatic acyl radical refers to C2-C6 alkanoyl (such as acetyl group, propionyl Base, positive bytyry, isobutyryl, positive valeryl, valeryl or positive caproyl), the substituted benzoyl refers to phenyl ring Replaced substituent group by one or more, the substituent group is selected from halogen (such as fluorine, chlorine, bromine or iodine), nitro, C1- C4 alkyl (such as methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group or tert-butyl), C1-C4 halogenated alkyl (such as Trifluoromethyl), C1-C4 alkoxy (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, n-butoxy, isobutoxy or Tert-butoxy)；Also, the substituent group is in any position of phenyl ring；Preferably, R₂And R₇Be each independently selected from acetyl group, Benzoyl, 4- chlorobenzoyl base, 4- bromobenzoyl base, are more preferably selected from acetyl group or benzoyl；

In formula G- formula O compound, R₃Selected from-(CH₂CH₂O)n-CH₂CH₂, n is 1-100 here；Aliphatic alkane subunit； Aliphatic olefin subunit；Or-(CH₂CH₂O)m-CH₂, m is 1-100 here；Preferably, it is selected from-CH₂CH₂-O-CH₂-；R₄It is selected from Azido, alkynyl, biotin, substituted amino, aldehyde radical, thioether；The substituted amino refer to by protecting group protect amino, The protecting group is selected from benzyloxy carbonyl acyl group, tertiary butyloxycarbonyl acyl group, trichloroacetyl, trifluoroacetyl group；Preferably ,-OR₃-R₄For- O-CH₂CH₂-O-CH₂-N₃；

In Formulas I, formula J, formula K, formula L and formula M compound, R₅It is here, described selected from unsubstituted phenyl or substituted phenyl Substituted phenyl refer to phenyl ring replaced one or more substituent group, the substituent group be selected from halogen (such as fluorine, chlorine, Bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group or tert-butyl), C1- C4 halogenated alkyl (such as trifluoromethyl), C1-C4 alkoxy (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, positive fourth Oxygroup, isobutoxy or tert-butoxy)；Also, the substituent group is in any position of phenyl ring；Preferably, phenyl, 4- first are selected from Phenyl, 4- chlorophenyl, 4- bromo phenyl, it is highly preferred that being selected from phenyl or 4- methoxyphenyl；

In formula L ', formula M ' and formula N ' compound, R₈And R₉It is each independently selected from C1-C4 alkyl, phenyl or substituted benzene Base；Here, the C1-C4 alkyl is selected from methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group or tert-butyl；It is described Substituted phenyl refer to phenyl ring replaced one or more substituent group, the substituent group be selected from halogen (such as fluorine, chlorine, Bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group or tert-butyl), C1- C4 halogenated alkyl (such as trifluoromethyl), C1-C4 alkoxy (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, positive fourth Oxygroup, isobutoxy or tert-butoxy)；Also, the substituent group is in any position of phenyl ring；Preferably, R₈And R₉It is respectively independent Ground is selected from methyl, isopropyl or tert-butyl or phenyl；

In formula E compound, X is selected from fluorine or chlorine, it is therefore preferable to chlorine；

In formula A- formula B compound, M is sodium, potassium, lithium or calcium, it is therefore preferable to sodium.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (1), hyaluronidase adds Entering the 2.0-3.0% that amount is Sodium Hyaluronate quality, reaction temperature is 37 DEG C, and the molecular weight of formula A compound is 1KDa-10, 000KDa, it is therefore preferable to 10KDa-1,000KDa；The faintly acid refers to that the pH value of buffer solution is 3.0-6.5, it is therefore preferable to 5.0-5.2；The buffer solution is selected from acetic acid-sodium acetate buffer solution, acetic acid -50 mM, sodium dihydrogen phosphate-phosphoric acid hydrogen Disodium buffer or potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer one kind, it is therefore preferable to from acetic acid-sodium acetate buffer solution；Institute The concentration for stating buffer is selected as 0.01M-1M；Preferably, the buffer solution is acetic acid-acetate buffer that concentration is 0.1M Liquid；Concentration of the sodium chloride in weak acid buffer solution is 0.05M-0.5M, it is therefore preferable to 0.15M；The hyaluronidase choosing From the hyaluronidase of animal testicular origin, such as the hyaluronic acid from the bull testis source that Sigma-Aldrich company buys Enzyme.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (2), the choosing of the inorganic acid From hydrogen chloride, sulfuric acid, nitric acid, phosphoric acid, it is preferable that be selected from hydrogen chloride or sulfuric acid；Alcohol R₁R in OH₁As described above；The alkalinity item Part refers to selected from sodium acetate, pyridine, sodium dihydrogen phosphate, potassium carbonate, condition existing for the alkali of triethylamine or piperidines, it is preferable that Refer to condition existing for the alkali selected from pyridine or sodium acetate.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (3), the alkali is selected from second two Amine, hydrazine hydrate or 3-N, N dimethyl amino propylamine, it is preferable that be selected from hydrazine hydrate or 3-N, N dimethyl amino propylamine；It is described to have Solvent be selected from acetone, chloroform, tetrahydrofuran, methylene chloride, n,N dimethylformamide or N, N dimethyl acetamide, preferably Ground is selected from methylene chloride, tetrahydrofuran or n,N dimethylformamide.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (4), the alkali is selected from alkali gold Carbonate, 1,8- diazabicylo, the 11 carbon -7- alkene (DBU) of category, acetate, borate, sodium hydroxide, in potassium hydroxide It is a kind of；Preferably, potassium carbonate or 1, one of 11 carbon -7- alkene (DBU) of 8- diazabicylo are selected from；Three haloacetonitrile In halogen be selected from fluorine, chlorine, bromine, it is therefore preferable to chlorine.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (5), the acid is selected from trifluoro Methanesulfonic acid alkyl silicone grease, boron trifluoride ether, copper chloride, p-methyl benzenesulfonic acid or camphorsulfonic acid.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (6), the acid is selected from trifluoro Methanesulfonic acid alkyl silicone grease, boron trifluoride ether, copper chloride, p-methyl benzenesulfonic acid, camphorsulfonic acid；Preferably, copper chloride, three are selected from The silicone grease that fluorine methanesulfonic acid C1-C4 trialkyl replaces；The organic solvent is selected from methylene chloride, chloroform, tetrahydrofuran, acetone or 1, One of 2- dichloroethanes；Preferably chloroform.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (7), the acid is selected from chlorination One of hydrogen, sulfuric acid, nitric acid, phosphoric acid or boron trifluoride ether, it is preferable that be selected from hydrogen chloride or boron trifluoride ether；It is described Alkali is selected from R₁ONa、R₁OK or NaOH, it is therefore preferable to R₁ONa, wherein R₁Selected from unsubstituted benzyl or substituted benzyl, C1-C4 Alkyl or allyl；Here, the substituted benzyl refer to phenyl ring replaced one or more substituent group, it is described to take Dai Ji is selected from halogen (such as fluorine, chlorine, bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-propyl, isopropyl, just Butyl, isobutyl group or tert-butyl), C1-C4 halogenated alkyl (such as trifluoromethyl), C1-C4 alkoxy (such as methoxyl group, ethoxy Base, positive propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy)；Also, the substituent group is in any of phenyl ring Position；Preferably, R₁Selected from methyl, ethyl, allyl, benzyl, 4- methoxy-benzyl is more preferably selected from methyl, ethyl, benzyl Base.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (8), the acid is selected to first Base benzene sulfonic acid or camphorsulfonic acid.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (9), the alkaline condition refers to Be selected from sodium acetate, pyridine, sodium dihydrogen phosphate, potassium carbonate, triethylamine or piperidines alkali existing for condition, it is preferable that refer to It is condition existing for the alkali selected from pyridine or sodium acetate.

In the synthetic method of chondroitin sulfate provided by the present application, in the step (10), the acid is selected from trifluoro second One of acid, acetic acid, boric acid, it is therefore preferable to trifluoroacetic acid；The reducing agent is selected from silane (such as three second that trialkyl replaces Base silane, tri isopropyl silane, t-butyldimethyl silane, pheiiyldimetliyl silane), sodium cyanoborohydride, borine；It is preferred that Ground, the silane replaced selected from trialkyl (such as triethylsilane, tri isopropyl silane, t-butyldimethyl silane, phenyl diformazan Base silane) or sodium cyanoborohydride.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (11), the oxidant choosing From Dai Si-Martin's oxidant, the combination of acid anhydrides-dimethyl sulfoxide, the combination of the chloro- dimethyl sulfoxide-triethylamine of oxalyl, trifluoro second Acid anhydrides-dimethyl sulfoxide-triethylamine combination or manganese dioxide；It preferably, is Dai Si-Martin's oxidant；

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (12), the reducing agent choosing From sodium borohydride, lithium borohydride, potassium borohydride, alkyl-substituted borohydride salts (such as: triisobutyl potassium borohydride or three isobutyls Base lithium borohydride)；Preferably, triisobutyl potassium borohydride or sodium borohydride are selected from.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (13), the oxidant choosing From DDQ or ammonium ceric nitrate；Combination or triethylsilane and carbon monoxide of the reducing agent selected from triethylsilane and iodine With the combination of cobalt octacarbonyl.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (10a), the acid is selected from three One of fluoroacetic acid, acetic acid, boric acid, it is preferable that be acetic acid.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (11a), the substituted chlorine Silane R₈R₈R₉SiCl is selected from tert-butyl diphenyl chlorosilane, tert-butyl chloro-silicane, tri isopropyl chlorosilane, triethyl group Chlorosilane, 3,5-dimethylphenyl chlorosilane or trim,ethylchlorosilane；It preferably, is tert-butyl chloro-silicane；The alkali is selected from Triethylamine, pyridine or 4-dimethylaminopyridine, it is preferable that be pyridine.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (12a), the oxidant choosing From the combination of the chloro- dimethyl sulfoxide-triethylamine of oxalyl, the combination of acetic anhydride-dimethyl sulfoxide, manganese dioxide or Dai Si-Martin High iodine reagent；The preferably high iodine reagent of Dai Si-Martin.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (13a), the reducing agent choosing From sodium borohydride, lithium borohydride, potassium borohydride or alkyl-substituted borohydride salts (such as: triisobutyl potassium borohydride is three different Butyl lithium borohydride)；Preferably, triisobutyl potassium borohydride or sodium borohydride are selected from.

In the synthetic method of chondroitin sulfate tetrose provided by the present application, in the step (14), it is described acid selected from pair Toluenesulfonic acid or trifluoroacetic acid, preferably p-methyl benzenesulfonic acid；The fluorine ion reagent is selected from tetrabutyl ammonium fluoride, hydrogen fluoride pyridine Complex compound, hydrogen fluoride triethylamine complex or ammonium fluoride；Preferably hydrogen fluoride pyridine complex compound.

The synthetic method of chondroitin sulfate tetrose provided by the present application, after obtaining formula O compound, further include as One of lower step:

(i) 1 equivalent formula O compound is in the presence of the sulfur trioxide organic base compound of 20-100 equivalent to hydroxyl thereon Base carries out sulfonic acid esterification, then sloughs protecting group under alkaline condition and obtains chondroitin sulfate E tetrose, i.e. formula CS-E compound；

(ii) 1 equivalent formula O compound is in the presence of the sulfur trioxide organic base compound of 5-15 equivalent to N- acetyl ammonia On base galactose residue 6 and 6 ' two hydroxyls of position carry out the esterification of regioselectivity sulfonic acid, then slough under alkaline condition Protecting group obtains chondroitin sulfate C tetrose, i.e. formula CS-C compound；

(iii) formula O compound and Cyanophenacyl are under the action of organic base to 6 on N- acetylgalactosamine residues With 6 ' two hydroxyls of position carry out regioselectivity benzoylation, then in the presence of sulfur trioxide organic base compound to 4 with The hydroxyl of 4 ' positions carries out sulfonic acid esterification, then sloughs protecting group under alkaline condition and obtain chondroitin sulfate A (CSA) tetrose, i.e. formula CS-Aization Close object；

Here, step (i), that the sulfur trioxide organic base compound in (ii) and (iii) is selected from sulfur trioxide pyridine is compound Object, sulfur trioxide triethylamine compound or sulfur trioxide trimethylamine compound, it is preferable that be sulfur trioxide trimethylamine compound. The alkaline condition refers to condition existing for following alkali: sodium hydroxide or its mixed solution with hydrogen peroxide, potassium hydroxide Or itself and the mixed solution of hydrogen peroxide, the mixed solution of lithium hydroxide or itself and hydrogen peroxide, it is preferably chosen from lithium hydroxide Or the mixed solution of itself and hydrogen peroxide.

In addition, after obtaining formula O compound, can join in the synthetic method of chondroitin sulfate tetrose provided by the present application Examine the introduction (bibliography: Jean-Claude Jacquinet et al, From Polymer to Size- of the prior art Defined Oligomers:A Highly Divergent and Stereocontrolled Construction of Chondroitin Sulfate A,C,D,E,K,L,and M Oligomers from a Single Precursor:Part 2.Chem.Eur.J.2009,15,9579-9595.) prepare chondroitin sulfate tetrose A, C or E.

Present invention also provides the midbody compounds for synthesizing chondroitin sulfate tetrose, are selected from formula D- formula N ' compound One of:

Here, in formula D- formula N ' compound, Ac is acetyl group；

In formula D- formula N ' compound, R₁Selected from unsubstituted benzyl or substituted benzyl, C1-C4 alkyl or allyl；This In, the substituted benzyl refer to phenyl ring replaced one or more substituent group, the substituent group be selected from halogen (such as Fluorine, chlorine, bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group or tertiary fourth Base), C1-C4 halogenated alkyl (such as trifluoromethyl), C1-C4 alkoxy (such as methoxyl group, ethyoxyl, positive propoxy, isopropyl oxygen Base, n-butoxy, isobutoxy or tert-butoxy)；Also, optionally, the substituent group is in any position of phenyl ring；It is preferred that Ground, R₁Selected from methyl, ethyl, allyl, benzyl, 4- methoxy-benzyl is more preferably selected from methyl, ethyl, benzyl；

In formula D- formula N ' compound, R₂And R₇It is each independently selected from aliphatic acyl radical, unsubstituted benzoyl or substitution Benzoyl；Here, the aliphatic acyl radical refers to C2-C6 alkanoyl (such as acetyl group, propiono, positive bytyry, isobutyl Acyl group, positive valeryl, valeryl or positive caproyl)；The substituted benzoyl refers to phenyl ring by one or more Substituent group is replaced, the substituent group be selected from halogen (such as fluorine, chlorine, bromine or iodine), nitro, C1-C4 alkyl (such as methyl, Ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group or tert-butyl), C1-C4 halogenated alkyl (such as trifluoromethyl), C1-C4 alkane Oxygroup (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy)；Also, appoint Selection of land, the substituent group is in any position of phenyl ring；Preferably, R₂And R₇It is each independently selected from acetyl group, benzoyl, 4- Chlorobenzoyl base, 4- bromobenzoyl base, are more preferably selected from acetyl group or benzoyl；

In formula G- formula N ' compound, R₃Selected from-(CH₂CH₂)n-O-CH₂CH₂, n is 1-100 here；Aliphatic alkane is sub- Base；Aliphatic olefin subunit；Or-(CH₂CH₂)n-O-CH₂, n is 1-100 here；Preferably, it is selected from-CH₂CH₂-O-CH₂-；R₄ Selected from azido, alkynyl, biotin, substituted amino, aldehyde radical, thioether；The substituted amino refers to by protecting group protection Amino, the protecting group are selected from benzyloxy carbonyl acyl group, tertiary butyloxycarbonyl acyl group, trichloroacetyl, trifluoroacetyl group；Preferably, OR₃-R₄ For-O-CH₂CH₂-O-CH₂-N₃；

In Formulas I, formula J, formula K, formula L and formula M compound, R₅It is here, described selected from unsubstituted phenyl or substituted phenyl Substituted phenyl refer to phenyl ring replaced one or more substituent group, the substituent group be selected from halogen (such as fluorine, chlorine, Bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group or tert-butyl), C1- C4 halogenated alkyl (such as trifluoromethyl), C1-C4 alkoxy (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, positive fourth Oxygroup, isobutoxy or tert-butoxy)；Also, optionally, the substituent group is in any position of phenyl ring；Preferably, it is selected from benzene Base, 4- methoxyphenyl, 4- chlorophenyl, 4- bromo phenyl, it is highly preferred that being selected from phenyl, 4- methoxyphenyl；

In formula L ', formula M ' and formula N ' compound, R₈And R₉It is each independently selected from C1-C4 alkyl, phenyl or substituted benzene Base；Here, the C1-C4 alkyl refers to methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group or tert-butyl；It is described Substituted phenyl refer to phenyl ring replaced one or more substituent group, the substituent group be selected from halogen (such as fluorine, chlorine, Bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group or tert-butyl), C1- C4 halogenated alkyl (such as trifluoromethyl), C1-C4 alkoxy (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, positive fourth Oxygroup, isobutoxy or tert-butoxy)；Also, optionally, the substituent group is in any position of phenyl ring；Preferably, R₈And R₉ It is each independently selected from methyl, isopropyl, tert-butyl.

In formula E compound, X is selected from fluorine or chlorine, it is therefore preferable to chlorine.

The midbody compound provided by the present application for being used to synthesize chondroitin sulfate tetrose, comprising:

(tri--O- acetyl group of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group of 4,6-- 2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β-D- glucopyra alditol of 2,3- Sour carbomethoxy) two-O- acetyl group -2- '-deoxy-n of-(1 → 3) -4,6--acetylaminohydroxyphenylarsonic acid D- glucopyranose；

O- (((tri--O- acetyl group of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(bis--O- second of 4,6- Acyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β-D- glucopyra of 2,3- Uronic acid carbomethoxy)-(1 → 3) -) two-O- acetyl group -2- '-deoxy-n of-O-4,6--acetylaminohydroxyphenylarsonic acid D- glucopyranosyl) three Chloroethene imide ester；

O- (tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- of 4,6- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β of 2,3--D-Glucose aldehydic acid methyl esters Base)-(1 → 3) -2- methyl-(bis--O- acetyl group -1,2- dideoxy-α-D- glucopyranose of 4,6-) [2,1-d] 2- oxazoline；

1- nitrine -2- oxa- -4-O- [(tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid methyl esters)-(1 → 3)-(4, Bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl of 6-)-(1 → 4)-(bis--O- acetyl group of 2,3- - β-D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- pyrans Portugal 4,6- Grape glycosyl)] butane；

1- nitrine -2- oxa- -4-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetyl Amino-beta-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- Acetylaminohydroxyphenylarsonic acid α-D- glucopyranosyl)] butane；

[(4,6-O- is to methoxy benzal for (beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-by 1- nitrine -2- oxygen -4-O- Base -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to benzylidene -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] butane；

1- nitrine -2- oxa- -4-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- Two-O- benzoyls-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- deoxidation -2- N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] butane；

1- nitrine -2- oxa- -4-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- of 2,3- Benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -2- deoxidation -2-N- acetyl ammonia Base-β-D- glucopyranosyl)] butane；

1- nitrine -2- oxa- -4-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -4- ketone -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- Two-O- benzoyls-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -4- ketone -2- deoxidation -2- N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] butane；

1- nitrine -2- oxa- -4-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(bis--O- of 2,3- Benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -2- deoxidation -2-N- acetyl ammonia Base-β-D- galactopyranosyl glycosyl)] butane；

1- nitrine -6-O- [(tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid methyl esters)-(1 → 3)-(bis--O- of 4,6- Acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β-D- grape of 2,3- Uronic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- glucopyranosyl of 4,6-)] Hexane；

1- nitrine -6-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β - D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetyl ammonia Base-α-D- glucopyranosyl)] hexane；

1- nitrine -6-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- benzal -2- deoxidation -2- N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- Benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane；

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4, 6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl-β of 2,3- - D- glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyra Glycosyl)] hexane；

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- Deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl-β-D- glucopyranose of 2,3- Aldehydic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane；

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6- O- dimethyl tertiary butyl silicon ether -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- of 2,3- Benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -2- deoxidation -2-N- Acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane；

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6- O- dimethyl tertiary butyl silicon ether -4- ketone -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(2,3- Two-O- benzoyls-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -4- ketone - 2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane；

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6- O- dimethyl tertiary butyl silicon ether -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(bis--O- of 2,3- Benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -2- deoxidation -2-N- Acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane；With

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- Deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(bis--O- benzoyl-β-D- glucopyranose of 2,3- Aldehydic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane.

The application using it is more easy to produce, be easier to extract obtained Sodium Hyaluronate as raw material, by cheap animal testis come Raw material is hydrolyzed in the hyaluronidase in source, the tetrose skeleton being chemically modified after obtaining can be used for, then to its into The conventional protecting group operation of row can be used for chondroitin sulfate A (CSA) to synthesis, and the key intermediate of tri- kinds of tetroses of C, E synthesis is whole For route without carrying out glycosylation operation, reaction route is short, high income, and can have azido, reacts convenient for later-stage utilization click It is marked, be produced on a large scale chondroitin sulfate and its derivative；On the other hand, pass through condition optimizing appropriate and purifying side Formula selection can obtain the higher sample of purity for studying, explore convenient for the medicineization of chondroitin sulfate.

Specific embodiment

The exploitativeness of the application is further explained below by embodiment, not to the limit of the application protection scope System.

Detecting instrument:

Nuclear-magnetism: Bruker AV-400 type Nuclear Magnetic Resonance, solvent CDCl₃,CD₃OD, TMS are internal standard.

Mass spectrum: Bruker APEX IV type mass spectrograph.

Abbreviation:

CH₂Cl₂For methylene chloride

EtOAc is ethyl acetate

MeOH is methanol

DMF is N,N-dimethylformamide

DMP: Dai Si-Martin reagent

HRMS: high resolution mass spectrum

ESI: electrospray ionization mass spectrum

Embodiment 1:(beta d glucopyranosiduronic acid base)-(1 → 3)-(2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyra Glycosyl)-(1 → 4)-(beta d glucopyranosiduronic acid base)-(1 → 3) -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid D- glucopyranose disodium The synthesis of salt

Hyaluronic acid dry powder (molecular weight about 500,000) 51.0g is taken to be placed in the sodium chloride containing 0.15M of the pH=5.00 of 2500mL 0.10M sodium acetate-acetic acid buffer in, make with vigorous stirring hyaluronic acid at room temperature sufficiently swelling (about needing 72h) be Colourless, uniform sticky colloidal liquid.So that the colloidal liquid is warming up to 37.0 DEG C using water bath with thermostatic control, it is transparent that 1.0g is added at this time Matter acid enzyme dry powder (stir at 37.0 DEG C from Sigma-Aldrich company, article No. H-3506, Lot#SLBL1922V) by purchase Two weeks.Reaction terminates for reaction solution to be warming up to boiling, and in 105 DEG C of stirring 15min, after solution does not continue to muddiness rapidly by it It is cooled to room temperature, and is evaporated under reduced pressure after being mixed with the dehydrated alcohol of 1/20 volume, be concentrated to dryness to obtain white syrup.Later by this sugar Slurry, which is dissolved and is slowly dropped into the absolute ethanol that 3000mL is vigorously stirred with a small amount of distilled water, is precipitated white precipitate, to be filtered under diminished pressure Method precipitating be precipitated is collected in environment of the relative air humidity no more than 50%, and washed with absolute ethanol cold on a small quantity Filter cake is washed, obtained solid powder will be then collected and is dried under reduced pressure in the drier for filling anhydrous calcium chloride to constant weight, obtained 96.0g white amorphous powder shape solid.The crude product can be directly used for lower step synthesis, without being further purified.It is anti-to determine The product structure and yield answered, 200mg crude product are pure through LH-20 sephadex column (pillar height × column diameter=100cm × 2cm) Change, to distill water elution, title tetrose product 60mg is obtained, with document P.Litant et al.Carbohydr.Res.237 (1992) 271-281 is provided¹H NMR spectra is more identical.

Tri--O- acetyl group of embodiment 2:(2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6- bis-- O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β-D- pyrans of 2,3- Glucuronic acid methyl ester base) conjunction of three-O- acetyl group -2- '-deoxy-n of-(1 → 3) -1,4,6--acetylaminohydroxyphenylarsonic acid D- glucopyranose At

By (beta d glucopyranosiduronic acid base)-(1 → 3)-(2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)- (1 → 4)-(beta d glucopyranosiduronic acid base)-(1 → 3) -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid D- glucopyranose disodium salt crude product 96.0g is dissolved in 12L, in the hydrogen chloride methanol solution of 0.08M newly to freeze.Then by reaction solution in 4 DEG C of standing 96h.Reaction knot Reaction solution is neutralized to neutrality with triethylamine after beam, is then evaporated under reduced pressure, and be concentrated to dryness.Pyridine 405mL is added, is cooled to 0 DEG C, it is slowly added dropwise after acetic anhydride 230mL and reaction solution is slowly increased to room temperature, and stir at room temperature for 24 hours.To after reaction, Reaction solution is cooling in ice-water bath, methanol 170mL is slowly added dropwise, and in stirring 1h at this temperature, reaction solution is evaporated under reduced pressure It is concentrated to give brown color syrup.This syrup is dissolved in 2.5L EtOAc, removing insoluble matter is filtered under diminished pressure, organic phase is successively used The 1M aqueous hydrochloric acid solution of 400mL washs 3 times, and water phase is extracted three times with 400mL EtOAc.Merge organic phase, and successively uses 600mL Saturated sodium bicarbonate aqueous solution washs 2 times, 600mL saturated common salt water washing 1 time.By organic phase anhydrous sodium sulfate drying, mistake Filter, vacuum distillation, gained crude product methylene chloride: ethyl acetate: (eluant, eluent contains methanol=1:1:0.06 to 1:1:0.14 0.1% triethylamine) Flash silica column chromatographic purifying, obtain 32.6g white, amorphous solid.Sodium Hyaluronate from embodiment 1 For raw material, three step yields 42%.

R_f=0.38 (SiO₂,CH₂Cl₂: EtOAc:MeOH=1:1:0.18).

¹H NMR shows that product is the mixture of α: β=3:1.

HRMS(ESI):[M+H]⁺Calculated value 1225.3777, measured value [M+H]⁺: 1225.3750.

Tri--O- acetyl group of embodiment 3:(2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6- bis-- O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β-D- pyrans of 2,3- Glucuronic acid methyl ester base) synthesis of two-O- acetyl group -2- '-deoxy-n of-(1 → 3) -4,6--acetylaminohydroxyphenylarsonic acid D- glucopyranose

Take (tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- of 4,6- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranose)-(1 → 4)-(bis--O- acetyl group-β of 2,3--D-Glucose aldehydic acid methyl esters Base) three-O- acetyl group -2- '-deoxy-n of-(1 → 3) -1,4,6--acetylaminohydroxyphenylarsonic acid D- glucopyranose 30.2g (24.6mmol) is molten In 240mL tetrahydrofuran, it is slowly added dropwise 15.5mL (90.7mmol) 3-N, N- dimethyl-amino propylamine under stiring, and in 3h is stirred at room temperature.Reaction solution 650mL chloroform is diluted, organic phase is washed 3 times with the 1M hydrochloric acid solution of 150mL, and water phase is used 150mL chloroform extracts 3 times, merges organic phase, is washed 1 time with 230mL saturated sodium bicarbonate aqueous solution, saturated common salt water washing 1 Secondary, organic phase is dried, filtered with anhydrous sodium sulfate, and the faint yellow amorphous powder of 25.5g, yield 88% is concentrated under reduced pressure to obtain.It is thick to produce Object can be used to following reaction without being further purified.

Embodiment 4:O- (((tri--O- acetyl group of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6- Two-O- acetyl group -2- '-deoxy-ns-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β-D- of 2,3- Glucopyranosiduronic acid carbomethoxy)-(1 → 3) -) two-O- acetyl group -2- '-deoxy-n of-O-4,6--acetylaminohydroxyphenylarsonic acid D- glucopyra Glycosyl) tri- chloroacetimidate synthesis

Take dry (tri--O- acetyl group of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6- bis-- O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β-D- pyrans of 2,3- Glucuronic acid methyl ester base) two-O- acetyl group -2- '-deoxy-n of-(1 → 3) -4,6--acetylaminohydroxyphenylarsonic acid D- glucopyranose crude product 7.0g (6.0mmol) is dissolved in 60mL dry methylene chloride, is added Tritox 6.7mL (60mmol), by this mixture in argon It is cooling in ice-water bath under gas atmosphere.11 carbon -7- alkene (DBU) of 1,8- diazabicylo, 149 μ L is slowly added dropwise later (0.9mmol).Reaction solution stirs 2.5h at 0 DEG C.To which reaction solution is concentrated under reduced pressure after the reaction was completed, and with the quick column color of silica gel Spectrum purifying (methylene chloride: ethyl acetate: methanol=3:1:0.10 contains 0.1% triethylamine) obtains 5.5g white, amorphous solid, receives Rate 79%.

R_f=0.48 (SiO₂,CH₂Cl₂: EtOAc:MeOH=1:1:0.12)；

¹H NMR(400MHz,CDCl₃) δ 8.83 (s, 1H, C=NH), 6.23 (d, J=3.7Hz, 1H), 5.84 (d, J= 7.5Hz, 1H), 5.77 (d, J=9.6Hz, 1H), 5.21 (dd, J=J=9.3Hz, 1H), 5.15-5.07 (m, 3H), 4.90 (dd, J=J=9.4Hz, 1H), 4.83-4.73 (m, 4H), 4.63 (d, J=7.8Hz, 1H), 4.59 (ddd, J=10.1Hz, 3.8Hz, 3.8Hz, 1H), 4.43 (dd, J=J=9.7Hz, 1H), 4.29 (dd, J=4.2Hz, 12.4Hz, 1H), 4.18 (dd, J =4.2Hz, 12.5Hz, 1H), 4.12-3.85 (m, 11H), 3.72 (s, 3H), 3.66-3.64 (m, 1H), 3.14 (ddd, J= 17.4Hz,8.0Hz,8.0Hz,1H),2.08-2.01(m,33H,COCH₃)。

¹³C NMR(100MHz,CDCl₃)δ171.0,170.8,170.2,170.1,169.8,169.8,169.7,169.4, 169.2,169.1,167.9,167.0,160.2,100.9,100.0,98.7,95.3,90.8,77.3,76.8,75.8,74.9, 72.3,72.1,71.8,71.8,71.7,71.6,70.2,69.4,68.0,67.7,61.8,61.7,57.7,53.0,52.8, 51.8,23.6,23.2,20.8,20.7,20.6,20.6,20.5,20.5。

HRMS(ESI):[M+H]⁺Calculated value 1326.2768, measured value [M+H]⁺: 1326.2738.

Embodiment 5:O- (tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(bis--O- of 4,6- Acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β of 2,3--D-Glucose Aldehydic acid carbomethoxy)-(1 → 3) -2- methyl-(bis--O- acetyl group -1,2- dideoxy-α-D- glucopyranose of 4,6-) [2,1-d] 2- oxazoline

Take dry O- (((tri--O- acetyl group of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6- Two-O- acetyl group -2- '-deoxy-ns-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β-D- of 2,3- Glucopyranosiduronic acid carbomethoxy)-(1 → 3) -) two-O- acetyl group -2- '-deoxy-n of-O-4,6--acetylaminohydroxyphenylarsonic acid D- glucopyra Glycosyl) tri- chloroacetimidate 9.0g (6.8mmol) is dissolved in 80mL dry methylene chloride, and in ice-water bath under argon atmosphere Middle cooling.182 μ L (1.02mmol) of Trimethylsilyl trifluoromethanesulfonate is slowly added dropwise.Reaction solution stirs 0.5h at 0 DEG C.To anti- 150 μ L triethylamine quenching reactions should be added dropwise after the completion.Reaction solution is concentrated to dryness, and with silica gel flash column chromatography (methylene chloride: ethyl acetate: methanol=3:1:0.15 contains 0.1% triethylamine) obtains 7.1g white, amorphous solid, yield 90%.

R_f=0.34 (SiO₂,CH₂Cl₂: EtOAc:MeOH=1:1:0.18)；

¹H NMR(400MHz,CDCl₃) δ 5.92 (d, J=6.6Hz, 1H), 5.82 (d, J=7.3Hz, 1H), 5.20 (dd, J =J=9.3Hz, 1H), 5.12 (dd, J=J=9.0Hz, 1H), 5.06 (dd, J=J=9.2Hz, 1H), 4.92-4.78 (m, 6H), 4.65 (d, J=7.8Hz, 1H), 4.60 (d, J=7.8Hz, 1H), 4.52 (m, 1H), 4.41-4.30 (m, 2H), 4.20 (dd, J=4.2Hz, 12.5Hz, 1H), 4.12-3.92 (m, 6H), 3.82 (s, 3H), 3.71-3.63 (m, 11H), 3.41 (m, 2H),3.30(m,1H),3.01(m,1H),2.06-1.96(m,33H).

¹³C NMR(100MHz,CDCl₃)δ171.2(2C),170.8,170.2,170.1,169.8,169.8,169.7, 169.4,169.2,169.1,167.9,167.0,100.7,100.0,99.4,98.3,77.9,76.7,75.6,74.5,72.3, 72.2,72.2,71.9,71.8,71.7,71.6,70.4,69.9,69.4,68.9,68.5,68.1,62.4,62.0,58.1, 57.5,52.9,52.7,50.7,23.6,23.4,20.8,20.7,20.6,20.6,20.5,20.5；

HRMS (ESI): calculated value [M+H]⁺: 1165.3566, measured value [M+H]⁺: 1165.3561.

Embodiment 6:1- nitrine -2- oxa- -4-O- [(tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid methyl esters)-(1 → 3)-(bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl of 4,6-)-(1 → 4)-(bis--O- of 2,3- Acetyl group-β-D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- of 4,6- Glucopyranosyl)] butane

Take dry O- (tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(bis--O- of 4,6- Acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β of 2,3--D-Glucose Aldehydic acid carbomethoxy)-(1 → 3) -2- methyl-(bis--O- acetyl group -1,2- dideoxy-α-D- glucopyranose of 4,6-) [2,1-d] 2- oxazoline 5.0g (4.3mmol) is dissolved in the dry chloroform of 35mL, and -4 butanol of 1- nitrine -2- oxa- is added, and (it is prepared Method is referring to document: Johnson, Charles L.and Guo, Zhongwu, Journal of Carbohydrate Chemistry,32(5-6),301-323；2013) 5.6g (43mmol) and anhydrous cupric chloride 631mg (4.7mmol).Reaction solution 10mL distilled water is added after reflux 10h at 65 DEG C and continues the 1h that flows back.Reaction solution is cooled down at room temperature, unsaturated carbonate is added Hydrogen sodium water solution is simultaneously vigorously stirred.Water phase is extracted 3 times with 10mL methylene chloride, merges organic phase, is washed with saturation NaCl aqueous solution It washs, anhydrous sodium sulfate dries, filters, and is concentrated to give colourless syrup.It disperses the syrup in 30mL anhydrous ether and is vigorously stirred 12h, it is seen that a large amount of white solids are precipitated, and solid are collected by filtration, by solid silica gel flash column chromatography (methylene chloride: second Acetoacetic ester: methanol=1:1:0.10 contains 0.1% triethylamine) obtain 4.7g white, amorphous solid, yield 84%.

R_f=0.40 (SiO₂,CH₂Cl₂: EtOAc:MeOH=1:1:0.17)；

¹H NMR(400MHz,CDCl₃) δ 6.08 (m, 2H, NHAc), 5.20 (dd, J=J=9.3Hz, 1H), 5.12 (dd, J =J=9.0Hz, 1H), 5.06 (dd, J=J=9.2Hz, 1H), 4.92-4.78 (m, 6H), 4.65 (d, J=7.8Hz, 1H), 4.60 (d, J=7.8Hz, 1H), 4.52 (m, 1H), 4.41-4.30 (m, 2H), 4.20 (dd, J=4.2Hz, 12.5Hz, 1H), 4.12-3.92(m,6H),3.82(s,3H),3.71-3.63(m,11H),3.41(m,2H),3.30(m,1H),3.01(m,1H), 2.06-1.96(m,33H)。

¹³C NMR(100MHz,CDCl₃)δ171.2(2C),170.8,170.2,170.1,169.8,169.8,169.7, 169.4,169.2,169.1,167.9,167.0,100.7,100.0,99.4,98.3,77.9,76.7,75.6,74.5,72.3, 72.2,72.2,71.9,71.8,71.7,71.6,70.4,69.9,69.4,68.9,68.5,68.1,62.4,62.0,58.1, 57.5,52.9,52.7,50.7,23.6,23.4,20.8,20.7,20.6,20.6,20.5,20.5。

HRMS (ESI): calculated value [M+H]⁺: 1296.4261, measured value [M+H]⁺: 1296.4244.

Embodiment 7:1- nitrine -2- oxa- -4-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation - 2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl) (2- is de- for-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)- Oxygen -2-N- acetylaminohydroxyphenylarsonic acid α-D- glucopyranosyl)] butane

Take [(tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid methyl esters)-(1 → 3) -1- nitrine -2- oxa- -4-O- (bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl of 4,6-)-(1 → 4)-(bis--O- acetyl of 2,3- Base-β-D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- pyrans of 4,6- Glucosyl group)] butane 4.0g (3.1mmol) is dissolved in the hydrogen chloride methanol solution of 30mL 0.25M.Reaction solution stirs at room temperature 36h is mixed, to use " Amberlite " IRA-410OH after the reaction was completed^-Type anion exchange resin neutralization reaction liquid is to neutrality.It crosses It filters off and removes resin, resin is washed 3 times with methanol, and cleaning solution merges with filtrate to be concentrated to dryness, and is filling anhydrous calcium chloride Drier in it is dry, obtain 2.8g white, amorphous solid, yield 100%.Product can be used to subsequent without being further purified Reaction.

R_f=0.15 (SiO₂,CH₂Cl₂: MeOH=4:1).

Embodiment 8:1- nitrine -2- oxa- -4-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- Benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid methyl esters Base)-(1 → 3)-(4,6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] butane

Take 1- nitrine -2- oxa- -4-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- second Acylamino--β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2- N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] butane 2.0g (2.2mmol) is dissolved in the dry n,N dimethylformamide of 15mL, (+)-camphorsulfonic acid (0.22mmol) and P-methoxybenzal-dehyde dimethyl acetal (22mmol) is added, reaction solution is at reduced pressure conditions (0.5 atmospheric pressure) is in 50 DEG C of stirring 4h.Triethylamine quenching reaction is added, reaction solution evaporated under reduced pressure is obtained into yellow syrup.It should Syrup, which is scattered in 20mL anhydrous ether, is vigorously stirred 12h, it is seen that a large amount of white solids are precipitated, and solid is collected by filtration, and solid is used A small amount of cold diethyl ether solution washing containing 20% methanol, obtains 1.9g crude product, yield 75%.The crude product is without being further purified, i.e., It can be used for following reaction.

R_f=0.40 (SiO₂,CH₂Cl₂: MeOH=15:1)；

Embodiment 9:1- nitrine -2- oxa- -4-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid first Ester group)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl of 2,3--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- Deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] butane

Taking 1- nitrine -2- oxa- -4-O-, [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to methoxy Benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid methyl esters Base)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] butane 1.9g It is dissolved in 20mL pyrido and is cooled to 0 DEG C, chlorobenzoyl chloride is previously dissolved in 1mL pyridine, be then slowly added dropwise in reaction solution simultaneously In 0 DEG C of stirring 3h.Reaction solution evaporated under reduced pressure is diluted with ethyl acetate with saturated sodium bicarbonate aqueous solution quenching reaction, it is organic It is mutually successively washed with the aqueous hydrochloric acid solution of 0.5N, saturated sodium bicarbonate aqueous solution washing, saturation NaCl aqueous solution washing, anhydrous sulphur Sour sodium dries, filters, concentration.Concentrate precipitates to obtain 2.3g white, amorphous solid, yield: 85% through ether.

R_f=0.63 (SiO₂,CH₂Cl₂: MeOH=25:1).

HRMS: calculated value [M+Na]⁺: 1696.5277, measured value [M+Na]⁺: 1696.5290.

[(tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid methyl esters)-(1 → 3)-embodiment 10:1- nitrine -6-O- (bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl of 4,6-)-(1 → 4)-(bis--O- acetyl of 2,3- Base-β-D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- pyrans of 4,6- Glucosyl group)] hexane

Take dry O- (tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(bis--O- of 4,6- Acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β of 2,3--D-Glucose Aldehydic acid carbomethoxy)-(1 → 3) -2- methyl-(bis--O- acetyl group -1,2- dideoxy-α-D- glucopyranose of 4,6-) [2,1-d] 2- oxazoline 10.0g (8.6mmol) is dissolved in the dry chloroform of 90mL, and 6- nitrine -1- hexanol (its synthetic method ginseng is added See T.M.Shaikh and A.Sudalai, Eur.J.Org.Chem.2010,3437-3444) 9.7g (69mmol) and anhydrous chlorine Change copper 1.26g (9.4mmol).Reaction solution is cooled to room temperature after 65 DEG C of reflux 10h under an argon atmosphere, and unsaturated carbonate hydrogen is added Sodium water solution is simultaneously vigorously stirred.Water phase is extracted 3 times with 10mL methylene chloride, merges organic phase, is washed with saturation NaCl aqueous solution, Anhydrous sodium sulfate dries, filters, and is concentrated to give colourless syrup.It disperses the syrup in 60mL anhydrous ether and is vigorously stirred 12h, it can See that a large amount of white solids are precipitated, be filtered under diminished pressure and collect solid powder be precipitated, filter cake is washed with anhydrous ether, is obtained after dry white Color pulverulent solids 10.6g, yield 95%.

R_f=0.49 (SiO₂,CH₂Cl₂: EtOAc:MeOH=1:1:0.17)=0.51；

(c1.0,CHCl₃)；

m.p.180-182℃(CH₂Cl₂/PE)；

¹HNMR(400MHz,CDCl₃) δ 5.92 (d, J=5.8Hz, 1H, GluNAc^III- NHAc), 5.73 (d, J=7.8Hz, 1H,GluNAc^I- NHAc), 5.20 (dd, J=J=9.3Hz, 1H, GlcA^IV- H-3), 5.12 (dd, J=J=9.7Hz, 1H, GlcA^IV- H-4), 5.06 (dd, J=J=8.5Hz, 1H, GlcA^II-H-3),4.94-4.79(m,6H,GluNAc^III-H-4, GluNAc^I-H-4,GlcA^II-H-2,GlcA^IV-H-2,GluNAc^I-H-1,GluNAc^III- H-1), 4.63 (d, J=7.8Hz, 1H, GlcA^II- H-1), 4.60 (d, J=7.9Hz, 1H, GlcA^IV- H-1), 4.55 (dd, J=J=9.8Hz, 1H, GluNAc^III-H- 3), 4.48 (dd, J=J=9.7Hz, 1H, GluNAc^I- H-3), 4.33 (dd, J=12.4Hz, 4.5Hz, 1H, GluNAc^III-H- 6a), 4.22 (dd, J=12.2Hz, 4.9Hz, 1H, GluNAc^I-H-6a),4.10-4.06(m,2H,GluNAc^I-H-6b, GlcA^II-H-4),4.01-3.95(m,3H,GlcA^IV-H-5,GlcA^II-H-5,GluNAc^III-H-6b),3.87-3.82(m, 4H,-COOCH₃,-OCH₂(CH₂)₅N₃),3.71(s,3H,-COOCH₃),3.65-3.63(m,2H,GluNAc^III-H-5, GluNAc^I- H-5), 3.46 (dt, J=9.6Hz, 6.4Hz, 1H ,-OCH₂(CH₂)₅N₃), 3.27 (t, J=6.8Hz, 2H ,-O (CH₂)₅CH₂N₃),3.09(m,1H,GluNAc^I-H-2),2.98(m,1H,GluNAc^III-H-2),2.07-1.96(m,33H,- COCH₃),1.59(m,4H,-OCH₂(CH₂)₄CH₂N₃),1.37(m,4H,-OCH₂(CH₂)₄CH₂N₃)ppm；

¹³CNMR(100MHz,CDCl₃)δ171.2,171.1,170.8,170.6,170.2,170.0,169.7,169.4, (169.3,167.0,168.9,167.8,166.9 C=O), 100.6,100.1,98.9,98.2,77.6,76.6,75.6, 74.5,72.3,72.2,71.9,71.8,71.8,69.8,69.4,68.9,68.2,62.4,62.0,58.3,58.2,52.9, 52.7,51.3,29.3,28.7,26.4,25.5,23.7,23.5,20.8-20.4(COCH₃)ppm；

HRMS(ESI):[M+H]⁺: calculated value 1307.4552, measured value 1307.4567.

Embodiment 11:1- nitrine -6-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- second Acylamino--β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2- N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane

Take 1- nitrine -6-O- [(tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid methyl esters)-(1 → 3)-(4,6- bis- - O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis- Portugal-O- acetyl group-β-D- 2,3- Grape uronic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- glucopyranose of 4,6- Base)] hexane 4.0g (3.1mmol) is dissolved in the hydrogen chloride methanol solution of 30mL 0.25M.Reaction solution stirs 36h at room temperature, To use " Amberlite " IRA-410OH after the reaction was completed^-Type anion exchange resin neutralization reaction liquid is to neutrality.Filtering removal Resin, resin are washed 3 times with methanol, and cleaning solution merges with filtrate to be concentrated to dryness, and in the drying for filling anhydrous calcium chloride It is dry in device, obtain 2.8g white, amorphous solid, yield 100%.Product can be used to subsequent reactions without being further purified.

Embodiment 12:1- nitrine -6-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- benzal - 2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane

Take 1- nitrine -6-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetyl ammonia Base-β-D- glucopyranosyl)] hexane 7.8g (6.9mmol) is dissolved in the dry n,N dimethylformamide of 68mL, is added (+)- Camphorsulfonic acid 1.27g (5.5mmol) and P-methoxybenzal-dehyde dimethyl acetal 18.6mL (124mmol), reaction solution is in decompression item (0.5 atmospheric pressure) is stirred overnight in 50 DEG C under part.Triethylamine quenching reaction is added, reaction solution evaporated under reduced pressure is obtained into yellow sugar Slurry.10mLCH is dispersed by the syrup₂Cl₂In, after be slowly added to 120mL anhydrous ether with vigorous stirring, it is seen that a large amount of whites Solid is precipitated, and is filtered under diminished pressure collection solid, and solid is successively washed with water and a small amount of cold methanol, obtains 7.12g crude product, yield 84%.The crude product can be used to following reaction without being further purified.

R_f=0.55 (SiO₂,CH₂Cl₂: MeOH=7:1)

¹H NMR(400MHz,DMSO-d₆) δ 7.82 (d, J=8.7Hz, 1H), 7.63 (d, J=8.4Hz, 1H), 7.41- 7.32 (m, 10H, PhH), 5.58 (s, 2H), 5.27 (d, J=5.6Hz, 1H), 5.11 (d, J=3.9Hz, 1H), 4.91 (s, 1H), 4.67-4.63 (m, 2H), 4.55 (d, J=8.1Hz, 1H), 4.45-4.41 (m, 2H), 4.36 (d, J=7.5Hz, 1H), 4.24-4.16(m,2H),3.95-3.88(m,2H),3.77-3.52(m,18H),3.44-3.33(m,6H),3.16-3.10(m, 1H),3.03-2.99(m,2H),1.81(s,6H),1.51-1.45(m,4H),1.30(m,4H)ppm.

¹³C NMR(100MHz,DMSO-d₆)δ170.2,170.1,169.7,168.8,138.1,138.1,128.9, 128.4,126.5,126.4,103.9,103.8,101.6,100.7,100.2,100.1,79.4,79.3,79.1,78.4 (2C),76.2(2C),74.2(2C),73.7(2C),71.9,69.1,68.2,68.0,66.2(2C),55.6(2C),52.7, 52.2,51.0,29.3,28.7,26.3,25.3,23.5,23.4ppm.

HRMS(ESI):[M+H]⁺: calculated value C₅₀H₆₈N₅O₂₃, 1106.4300, measured value 1106.4318；[M+Na]⁺: meter Calculation value C₅₀H₆₇N₅NaO₂₃, 1128.4119, measured value 1128.4127；[M+K]⁺: calculated value C₅₀H₆₇N₅KO₂₃, 1144.3858, it is real Measured value 1144.3830.

Embodiment 13:1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy) - (1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- Two-O- benzoyls-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- deoxidation -2- N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane

Taking 1- nitrine -6-O-, [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene - 2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane 5.0g (4.5mmol) is dissolved in 60mL pyrido and is cooled to -15 DEG C, and chlorobenzoyl chloride 2.8mL (23.9mmol) is slowly added dropwise into reaction solution In and in 0 DEG C of stirring 3h.Reaction solution evaporated under reduced pressure is diluted with ethyl acetate with saturated sodium bicarbonate aqueous solution quenching reaction, Organic phase is successively washed with the aqueous hydrochloric acid solution of 0.5N, saturated sodium bicarbonate aqueous solution washing, saturation NaCl aqueous solution washing, nothing Aqueous sodium persulfate dries, filters, concentration.Concentrate precipitates to obtain 6.3g white, amorphous solid, yield: 86% through ether.

R_f=0.62 (SiO₂,CH₂Cl₂: MeOH=20:1)

¹³C NMR(100MHz,CDCl₃)δ170.8,170.7,167.3,167.0,165.6,165.2,165.2,165.0, 164.8 (C=O), 137.4,137.1,133.6,133.4,133.3,129.8,129.8,129.6,129.1,12 9.0, 128.9,128.7,128.6,128.5,128.4,128.3,128.3,126.2,126.0(aromatic C),101.3(PhCH, 2C),100.6,100.4,99.1,98.2,80.7,80.5,76.2,74.4,72.8,72.3,72.1,71.8,71.7,70.1, 69.9,68.8,67.9,65.8,65.4,58.8,58.4,52.9,52.5,51.3,29.3,28.7,26.4,25.4,22.9, 22.6ppm；

HRMS(ESI):[M+H]⁺: calculated value C₈₅H₈₈N₅O₂₈, 1626.5610, measured value 1626.5626.

Embodiment 14:1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy) - (1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl-β-D- of 2,3- Glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane

Take [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -6-O- (4,6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl of 2,3- Base-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- pyrrole Glucopyranoside base)] hexane 8.0g in 75% acetic acid heated in water solution to 80 DEG C, and at a temperature of this stir 1h after will reaction Liquid is cooled to room temperature.It is evaporated under reduced pressure together with toluene 3 times.Residue is washed with anhydrous ether, is filtered under diminished pressure, and filter cake is collected, and is done 6.0g white, amorphous solid, yield: 85% are obtained after dry.

R_f(SiO₂,CH₂Cl₂: MeOH=10:0.7)=0.31；

¹H NMR(400MHz,MeOD-d₄) δ 7.96-7.29 (m, 25H), 6.05 (dd, J=J=9.6Hz), 5.66 (dd, J =J=7.9Hz), 5.59 (dd, J=J=9.7Hz), 5.49 (dd, J=9.6Hz, 7.8Hz), 5.30 (dd, J=J= 7.6Hz), 5.15 (d, J=7.7Hz), 5.07 (d, J=7.4Hz), 4.66 (d, J=9.8Hz), 4.59 (d, J=8.3Hz), 4.49 (dd, J=J=8.2Hz), 4.33-4.31 (m, 2H), 3.90-3.76 (m, 6H), 3.73-3.63 (m, 6H), 3.57- 3.50(m,2H),3.47-3.37(m,2H),3.31-3.18(m,4H),3.13-3.06(m,2H),1.56-1.46(m,4H), 1.47(s,3H),1.44(s,3H),1.37-1.30(m,4H)ppm；

¹³C NMR(100MHz,MeOD-d₄)δ171.7,171.6,168.2,167.7,165.6,165.4,165.3, (165.2,165.0 C=O), 133.4,133.3,133.2,133.2,129.7,129.6,129.4,129.4,129.3, 129.1,128.7,128.6,128.3,128.2(aromatic C),101.3,100.8,100.6,100.5,84.4,83.6, 76.1(C×2),75.3,74.3,73.0,72.4,72.3,71.9,71.5,69.9,69.2(C×2),69.0,61.3,61.2, 54.9,54.3,52.1,52.0,51.0,29.0,28.4,26.1,25.2,21.6,21.4ppm；

HRMS(ESI):[M+H]⁺: calculated value C₇₁H₈₀N₅O₂₈, 1450.4984, measured value 1450.4978；[M+Na]⁺: meter Calculation value C₇₁H₇₉N₅NaO₂₈, 1472.4804, measured value 1472.4752.

Embodiment 15:1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy) - (1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4) - (bis--O- benzoyl of 2,3--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether- 2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane

Take [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -6-O- (2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl-β-D- glucopyra of 2,3- Uronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane 1.8g and dimethyl Tertiary butyl chloride silane 785mg is dissolved in 28mL anhydrous pyridine, and reaction solution is stirred at room temperature overnight under an argon atmosphere.To thin layer It is after analysing Chromatogram display fully reacting, reaction solution is cooling in ice-water bath, methanol quenching reaction is added dropwise.Reaction solution is depressurized dense Contracting, residue with Ethyl acetate dilution, successively uses 1M aqueous hydrochloric acid solution, saturated sodium bicarbonate aqueous solution, saturated common salt water washing. Organic phase is dried, filtered with anhydrous sodium sulfate, and concentration, silica gel column chromatography separates (CH₂Cl₂: MeOH=100:1 → 90:1 → 80:1 → 40:1 → 25:1) obtain white, amorphous solid (1.8g, 86%).

R_f(SiO₂,CH₂Cl₂: MeOH=20:1.1)=0.51；

¹H NMR(400MHz,CDCl₃)δ7.96-7.23(m,25H,benzoyl aromatic proton),5.91(dd, J=J=9.5Hz, 1H), 5.70 (d, J=6.6Hz, 1H, GluNAc^III- NHAc), 5.62 (dd, J=J=9.6Hz, 1H), 5.52-5.47 (m, 2H), 5.31 (dd, J=J=7.6Hz, 1H), 5.25 (d, J=6.7Hz, 1H, GluNAc^I-NHAc),4.93 (d, J=8.2Hz, 1H, GluNAc^I- H-1), 4.87 (d, J=7.7Hz, 1H), 4.80 (d, J=8.2Hz, 1H, GluNAc^III- ), H-1 4.58 (dd, J=10.1Hz, 7.5Hz, 1H, GluNAc^III- H-3), 4.58 (dd, J=10.1Hz, 8.3Hz, 1H, GluNAc^I- H-3), 3.69 (s, 3H ,-COOMe), 3.61 (s, 3H ,-COOMe), 2.72 (ddd, J=8.3Hz, 8.3Hz, 6.7Hz,GluNAc^I- H-2), 2.65 (ddd, J=10.1Hz, 8.2Hz, 6.6Hz, GluNAc^III-H-2),0.88(s,9H,Si- C(CH₃)₃),0.87(s,9H,Si-C(CH₃)₃),0.04(s,9H,3×Si(CH₃)),0.02(s,3H,Si(CH₃))ppm；

¹³C NMR(100MHz,CDCl₃)δ171.5,171.1,167.5,166.9,165.5,165.3,165.2,164.8, 164.7 (C=O), 133.6,133.6,133.5,133.3,129.9,129.8,129.7,129.1,128.9,12 8.7, 128.7,128.6,128.5,128.5,128.4,128.3(aromatic C),100.9,100.8,98.2(GluNAc^I-C1), 97.2(GluNAc^III-C1),82.9(C×2,GluNAc^{I and} ^III-C3),76.2,74.4,74.2,72.4,71.9,71.8, 71.7,71.7,70.1,69.5,69.4,69.3,63.0,62.7,58.4(GluNAc^I-C2),58.3(GluNAc^III-C2), 53.1,53.1,51.3,29.3,28.7,26.4,25.9,25.8,25.5,23.3,23.1,18.4,18.2,-5.1 (TBDMS),-5.2(TBDMS),-5.2(C×2,(TBDMS))ppm；

HRMS(ESI):[M+H]⁺: calculated value C₈₃H₁₀₈N₅O₂₈Si₂, 1678.6714, measured value 1678.6676.

Embodiment 16:1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy) - (1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -4- ketone -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl of 2,3--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon Ether -4- ketone -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane

Take [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -6-O- (6-O- dimethyl tertiary butyl silicon ether -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis- - O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -2- deoxidation -2- N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane 3.0g (1.8mmol) is dissolved in methylene chloride (18mL), Dai Si-horse is added dropwise 18h is stirred in the dichloromethane solution 9.0mmol of the high iodine reagent of fourth, reaction at room temperature.Contain 10% sulphur to use after the reaction was completed The aqueous solution quenching reaction of sodium thiosulfate and 5% sodium bicarbonate, organic phase saturated common salt water washing, anhydrous sodium sulfate is dry, Filtering is concentrated under reduced pressure, and residue passes through a short silicagel column (CH₂Cl₂: MeOH=10:1 elution) and concentrate eluant to obtain 2.9g white Color amorphous solid, yield 95%.Through white, amorphous solid obtained by nuclear-magnetism and Mass Spectrometric Identification be product and its monohydrate and The mixture of dihydrate, by mixture, thermal dehydration can be changed into corresponding product under high vacuum condition.

R_f=0.60 (SiO₂,CH₂Cl₂: EtOAc:MeOH=1:1:0.10).

¹H NMR(400MHz,CDCl₃)δ8.01-7.27(m,25H),6.41(d,1H,NHAc),6.13(d,1H,NHAc), 5.87 (dd, J=J=9.1Hz, 1H), 5.68 (dd, J=J=9.6Hz, 1H), 5.58-5.55 (m, 3H), 5.45 (dd, J=J =7.11Hz, 1H), 5.34-5.29 (m, 2H), 5.06 (d, J=9.5Hz, 1H), 5.01-4.97 (m, 2H), 4.48 (d, J= 9.1Hz, 1H), 4.23 (d, J=9.9Hz, 1H), 4.15 (d, J=9.6Hz, 1H), 4.06-4.04 (m, 2H), 3.84 (s, 3H), 3.82-3.73(m,2H),3.69(m,1H),3.63(s,3H),3.49-3.38(m,3H),3.29-3.23(m,4H),1.94(s, 3H),1.89(s,3H),1.58-1.26(m,8H),0.87(s,9H),0.80(s,9H),0.06(s,3H),0.05(s,3H),- 0.01(s,3H),-0.07(s,3H)ppm；

¹³C NMR(100MHz,CDCl₃) δ 198.6 (2C, ketone), 172.0-164.9 (9C=O), 133.5-128.4 (aromatic C),99.5,98.9(2C),98.6,79.1,78.4,78.0,77.8,76.2,74.0,73.2,72.1,71.9, 71.7,69.64,69.56,61.7,60.5,60.4,53.1,53.0,51.3,29.2,28.7,26.4,25.82(tert- Butyl C),25.81(tert-Butyl C),25.4,23.5,23.3,18.3,18.2,-5.3(2C),-5.4,-5.5ppm；

HRMS(ESI):[M+NH₄]⁺: calculated value C₈₃H₁₀₇N₆O₂₈Si₂, 1691.6629, measured value 1691.6666.

Embodiment 17:1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy) - (1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(tri--O- benzoyl-β of 2,3,4- - D- glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane

Take [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -6-O- (6-O- dimethyl tertiary butyl silicon ether -4- ketone -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2, Bis--O- benzoyl of 3--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -4- ketone - 2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane 1.7g (1.0mmol) is dissolved in 40mL and is newly evaporated dry tetrahydro furan In muttering, and -78 DEG C are cooled under argon atmosphere, the tetrahydrofuran solution of commercially available 1M triisobutyl lithium borohydride is slowly added dropwise 2.2mL (2.2mmol), and 30min is reacted under -78 DEG C of argon atmospheres.Reaction solution is quenched with glacial acetic acid, then subtracts reaction solution Pressure concentration dissolves residue with ethyl acetate (40mL).Organic phase successively uses saturated sodium bicarbonate aqueous solution, saturated common salt washing It washs, water phase is extracted with dichloromethane, and merges organic phase, is dried, filtered with anhydrous sodium sulfate, be concentrated to give white amorphous powder. This powder is dissolved in a small amount of methylene chloride, ether is added dropwise under fast stirring to white precipitate is precipitated, and continues that ether is added dropwise extremely Precipitation is complete, and precipitating is collected by centrifugation and is washed with a small amount of ether, dry.

This precipitating is placed in 50mL plastic centrifuge tube, 10mL tetrahydrofuran is added and 10mL pyrido is cooled to 0 DEG C.It is slow Slow to be added dropwise HFPyridine 1mL, reaction solution is slowly increased to room temperature and .TLC for 24 hours is stirred at room temperature to show after the reaction was completed, Reaction solution is dissolved in Excess ethyl acetate, and successively uses 10%CuSO₄Aqueous solution, 0.01N EDETATE SODIUM saline solution and full And brine It, organic phase are dried, filtered with anhydrous sodium sulfate, obtain white, amorphous solid.Obtained solid is successively used into nothing Water-ethanol and recrystallisation from isopropanol obtain 910mg white powder, two step yields 62%.

R_f=0.43 (SiO₂,CH₂Cl₂: MeOH=20:1.6)

¹H NMR(400MHz,CDCl₃/CD₃OD=2/1) δ 7.95-7.27 (m, 25H, PhH), 5.99 (dd, J=J= 9.5Hz, 1H), 5.63-5.50 (m, 3H), 5.31 (dd, J=J=7.9Hz, 1H), 5.15 (d, J=7.9Hz, 1H), 5.05 (d, J=7.6Hz, 1H), 4.62 (d, J=8.7Hz, 1H), 4.54 (d, J=10.2Hz, 1H), 4.42-3.36 (m, 2H), 4.24 (d, J=8.2Hz, 1H), 4.10-4.07 (m, 2H), 3.99-3.97 (m, 1H), 3.91-3.70 (m, 9H), 3.61 (s, 3H), 3.49 (dd, J=J=6.1Hz, 1H), 3.44-3.38 (m, 1H), 3.30-3.15 (m, 5H), 1.54-1.49 (m, 4H), 1.31-1.25 (m,10H)ppm.

¹³C NMR(100MHz,CDCl₃/CD₃OD=2/1) δ 172.5-164.3 (9C=O), 135.2-128.9 (ArH), 101.7,101.5,101.3,100.8,81.2,80.4,75.9,74.6,74.4,74.3,73.4,72.4,72.0,71.8, 71.6,70.1,68.9,67.7,66.7,61.0,59.9,52.4,52.3,51.1,29.1,28.5,26.2,25.2,21.7, 21.5ppm.

HRMS(ESI):[M+H]⁺: calculated value 1450.4983, measured value 1450.5001.

(beta d glucopyranosiduronic acid base)-(1 → 3)-embodiment 18:1- nitrine -6-O- (4,6- di-sulfate base -2- Deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(beta d glucopyranosiduronic acid base)-(1 → 3)-(4, 6- di-sulfate base -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane, i.e. chondroitin sulfate E tetrose

Take [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -6-O- (2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(tri--O- benzoyl-β-D- pyrans Portugal 2,3,4- Grape uronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane 300mg (1.0equiv.) and sulfur trioxide trimethylamine compound (20equiv.) is dissolved in anhydrous DMF (4mL).Reaction solution is at 50 DEG C DEG C Reaction solution is cooled to room temperature after lower reaction 48h, reaction solution is added to the glass column of Sephadex LH-20 filling with dropper Upper end (packing volume be (2.5cm × 140cm)), with methylene chloride: methanol=1:1 eluent, gained flow point warp The ultraviolet 254nm monitoring of silica gel plate, has the flow point of fluorescence to merge concentration, obtains white foam.This foaming solid is dissolved in THF:H₂O In the mixed solution (10mL) of=3:1, -10 DEG C are cooled to, the aqueous solution of the 1M LiOH of brand-new and mixing for 35% hydrogen peroxide is added dropwise It closes solution (volume ratio 2:1,5mL), reaction solution, which stirs 1h at -10 DEG C and moves back to room temperature, to be continued to stir 8h.Liquid cooling will be reacted But to 0 DEG C.It is added methanol (8mL), the aqueous solution (4mL) of 4M NaOH slowly moves to room temperature afterwards, and 12h is stirred at room temperature.Instead It is neutralized to neutrality using resin cation, is filtered, multiple with distilled water rinse resin, merging filtrate, vacuum distillation is concentrated to dryness, Residue is eluted, Sephadex LH-20 column desalting and purifying with distilled water, collects the flow point for having product, be lyophilized after merging.? 200mg white foam solid, yield 80%.

(6- sulfate group -2- the deoxidation-of (beta d glucopyranosiduronic acid base)-(1 → 3)-embodiment 19:1- nitrine -6-O- 2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(beta d glucopyranosiduronic acid base)-(1 → 3)-(6- sulfuric ester Base -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane, i.e. chondroitin sulfate C tetrose

Take [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -6-O- (2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(tri--O- benzoyl-β-D- pyrans Portugal 2,3,4- Grape uronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane 300mg (1.0equiv.) and sulfur trioxide trimethylamine compound (5equiv.) is dissolved in anhydrous DMF (10mL).Reaction solution is at 40 DEG C Reaction solution is cooled to room temperature after reaction 48h, reaction solution is added to the glass column of Sephadex LH-20 filling with dropper Upper end (packing volume is (2.5cm × 140cm)), with methylene chloride: methanol=1:1 eluent, gained flow point is through silicon The ultraviolet 254nm monitoring of offset plate, has the flow point of fluorescence to merge concentration, obtains white foam.This foaming solid is dissolved in THF:H₂O= In the mixed solution (10mL) of 3:1, -10 DEG C are cooled to, the mixing of the aqueous solution and 35% hydrogen peroxide of the 1M LiOH of brand-new is added dropwise Solution (volume ratio 2:1,5mL), reaction solution, which stirs 1h at -10 DEG C and moves back to room temperature, to be continued to stir 8h.Reaction solution is cooling To 0 DEG C.It is added methanol (8mL), the aqueous solution (4mL) of 4M NaOH slowly moves to room temperature afterwards, and 12h is stirred at room temperature.Reaction It is neutralized to neutrality with resin cation, is filtered, multiple with distilled water rinse resin, merging filtrate, vacuum distillation is concentrated to dryness, will Residue is eluted with distilled water, Sephadex LH-20 column desalting and purifying, is collected the flow point for having product, is lyophilized after merging.? 150mg white foam solid, yield 69%.

(4- sulfate group -2- the deoxidation-of (beta d glucopyranosiduronic acid base)-(1 → 3)-embodiment 20:1- nitrine -6-O- 2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(beta d glucopyranosiduronic acid base)-(1 → 3)-(4- sulfuric ester Base -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane, i.e. chondroitin sulfate A (CSA) tetrose

Take [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -6-O- (2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(tri--O- benzoyl-β-D- pyrans Portugal 2,3,4- Grape uronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane 190mg (1.0equiv.) is stirred at room temperature for 24 hours in the presence of anhydrous pyridine with Cyanophenacyl (4.0equiv.).Under ice-water bath with It being concentrated to dryness after methanol quenching reaction, residue is precipitated with anhydrous ether, and precipitating is collected by filtration, is dissolved with ethyl acetate, It is successively washed with 1N aqueous hydrochloric acid solution, is saturated NaHCO₃Water-soluble washing, saturated common salt water washing, anhydrous sodium sulfate dry, filter, Weak yellow foam shape solid is concentrated under reduced pressure to obtain.This solid is dissolved in sulfur trioxide trimethylamine compound (10equiv.) anhydrous In DMF (2.5mL).Reaction solution is cooled to room temperature by reaction solution after reacting 12h at 50 DEG C, and reaction solution is added to one with dropper The upper end (packing volume is (2.5cm × 140cm)) of the glass column of Sephadex LH-20 filling, with methylene chloride: methanol= The eluent of 1:1, gained flow point are monitored through the ultraviolet 254nm of silica gel plate, are had the flow point of fluorescence to merge concentration, are obtained white bubble Foam.This foaming solid is dissolved in THF:H₂In the mixed solution (10mL) of O=3:1, -10 DEG C are cooled to, the 1M of brand-new is added dropwise The mixed solution (volume ratio 2:1,2mL) of the aqueous solution of LiOH and 35% hydrogen peroxide, reaction solution stirs 1h at -10 DEG C and moves back Continue to stir 8h to room temperature.Reaction solution is cooled to 0 DEG C.It is added methanol (4mL), the aqueous solution (2mL) of 4M NaOH slowly moves afterwards To room temperature, and 12h is stirred at room temperature.Reaction is neutralized to neutrality with resin cation, filtering, more with distilled water rinse resin Secondary, merging filtrate, vacuum distillation is concentrated to dryness, and residue is eluted with distilled water, Sephadex LH-20 column desalting and purifying, The flow point for having product is collected, is lyophilized after merging.Obtain 80mg white foam solid, yield 58%.

Claims

1. a kind of synthetic method of chondroitin sulfate tetrose, described method includes following steps:

(1) formula A compound obtains formula Bization in the weak acid buffer solution containing sodium chloride under the action of hyaluronidase Close object；Here, the hyaluronidase derives from animal testis, vigor 400-1000IU/mg；The molecular weight of formula A compound For 1KDa-10,000KDa；

(2) alcohol R of the formula B compound in inorganic acid₁Esterification is carried out in OH solution, and the carboxyl of formula B compound is esterified； Then utilize acid anhydrides R₂-O-R₂, to formula B compound, remaining all hydroxyl carries out acylated protection under alkaline condition, obtains formula C Compound；

(3) formula C compound in the presence of a base, 1 of selectively removing reducing end N-acetylglucosamine in organic solvent Hydroxyl obtains formula D compound；

(5) formula E compound obtains formula F compound under acid catalysis

(7) formula G compound is in acid or the alcohol R of alkali₁In OH solution, R all on formula G compound is sloughed₂, obtain formula H compound；

(8) formula H compound is in R₅CHO or R₅CHO contracts in the presence of two fatty alcohols, the selective protection formula H compound under acid catalysis 4,6 dihydroxy and 4 ', 6 ' dihydroxy obtain compound of formula I；

(9) compound of formula I is in carboxylic acid halides R₇X or acid anhydrides R₇-O-R₇Under the action of, it is acylated under alkaline condition, obtains formula J chemical combination Object；

Here, carboxylic acid halides R₇X in X is selected from chlorine, bromine or iodine；

(10) formula J compound is under the existence condition of acid and reducing agent, exposed 4 hydroxyls of selectivity, so that 6 are R₅-CH₂It protects Shield, obtains formula K compound；

(12) formula L compound is under reducing agent existence condition, selective two ketone carbonyls by 4 and 4 ' positions in formula L compound It is reduced to alcohol, and two hydroxyl configurations of the configuration of two newly-generated hydroxyls and step (11) Chinese style K are on the contrary, obtain gala The formula M compound of sugared configuration；

(13) formula M compound removes two benzyls or substituted benzyl of 6 and 6 ' positions, obtains under oxidant or reducing agent effect To tetrol, i.e. formula O compound；

Alternatively, following step (10a)-(14) in step (10)-(13) are replaced:

(10a) formula J compound hydrolyzes two benzals or substituted benzal, obtains formula K ' chemical combination under the existence condition of acid Object；

(11a) formula K ' compound is in substituted chlorosilane R₈R₈R₉In the presence of SiCl and alkali, regioselectivity to two N- second The 6 of acylamino- glucose, 6 ' position hydroxyls carry out silanization, obtain formula L ' compound；

(12a) formula L ' compound is under oxidant existence condition, selectively by two N- acetylaminos in formula L ' compound The hydroxyl of the 4 of glucose and 4 ' positions is oxidized to ketone carbonyl, obtains formula M ' compound；

(13a) formula M ' compound is three-dimensional by the ketone carbonyl of the 4 of N-acetylglucosamine and 4 ' positions under reducing agent existence condition Selectivity is reduced to axial bond hydroxyl, obtains formula N ' compound；

(14) formula N ' compound removes 6 and 6 ' positions in N- acetylgalactosamine residues in the presence of acid or fluorine ion reagent Silicon ether obtains tetrol, i.e. formula O compound；

(15) formula O compound synthesis chondroitin sulfate tetrose is utilized；

Here, in formula A- formula O compound, Ac is acetyl group；

Formula C- formula O compound and R₁In OH, R₁Selected from unsubstituted benzyl or substituted benzyl, C1-C4 alkyl or allyl； Here, the substituted benzyl refers to phenyl ring replaced one or more substituent group, and the substituent group is selected from halogen, nitre Base, C1-C4 alkyl, C1-C4 halogenated alkyl, C1-C4 alkoxy；Also, the substituent group is in any position of phenyl ring；

Formula C- formula O compound, formula J- formula O compound, acid anhydrides R₂-O-R₂, carboxylic acid halides R₇X and acid anhydrides R₇-O-R₇In, R₂And R₇Respectively Independently selected from aliphatic acyl radical, unsubstituted benzoyl or substituted benzoyl；Here, the aliphatic acyl radical refers to C2-C6 alkanoyl；The substituted benzoyl refers to replaced substituent group of the phenyl ring by one or more, the substitution Base is selected from halogen, nitro, C1-C4 alkyl, C1-C4 halogenated alkyl, C1-C4 alkoxy；Also, the substituent group is appointed phenyl ring Meaning position；

Formula G- formula O compound and alcohol R₄-R₃In OH, R₃Selected from-(CH₂CH₂O)n-CH₂CH₂, n is 1-100 here；Aliphatic alkane Hydrocarbon subunit；Aliphatic olefin subunit；Or-(CH₂CH₂O)m-CH₂, m is 1-100 here；R₄Selected from azido, alkynyl, biology Element, the amino replaced, aldehyde radical, thioether；The substituted amino refers to that the amino protected by protecting group, the protecting group are selected from benzyl Oxygen carbonyl acyl group, tertiary butyloxycarbonyl acyl group, trichloroacetyl, trifluoroacetyl group；

Formulas I, formula J, formula K, formula L, formula M compound, R₅CHO and R₅CHO contracts in two fatty alcohols, R₅Selected from unsubstituted phenyl or Substituted phenyl, here, the substituted phenyl refer to phenyl ring replaced one or more substituent group, the substituent group Selected from halogen, nitro, C1-C4 alkyl, C1-C4 halogenated alkyl, C1-C4 alkoxy；Also, the substituent group is in any of phenyl ring Position；

In formula L ', formula M ' and formula N ' compound, R₈And R₉It is each independently selected from C1-C4 alkyl, phenyl or substituted phenyl, Here, the substituted phenyl refers to phenyl ring replaced one or more substituent group, and the substituent group is selected from halogen, nitre Base, C1-C4 alkyl, C1-C4 halogenated alkyl, C1-C4 alkoxy；Also, the substituent group is in any position of phenyl ring；

In formula E compound, X is selected from fluorine or chlorine；

In formula A- formula B compound, M is sodium, potassium, lithium or calcium.

2. the method for claim 1, wherein animal testis described in the step (1) is bull testis or sheep testicle.

3. the method for claim 1, wherein

Formula C- formula O compound and R₁In OH, R₁Selected from methyl, ethyl, allyl, benzyl, 4- methoxy-benzyl；

Formula C- formula O compound, formula J- formula O compound, acid anhydrides R₂-O-R₂, carboxylic acid halides R₇X and acid anhydrides R₇-O-R₇In, R₂And R₇Respectively Independently selected from acetyl group, benzoyl, 4- chlorobenzoyl base, 4- bromobenzoyl base；

Formula G- formula O compound and alcohol R₄-R₃In OH ,-OR₃-R₄For-O-CH₂CH₂-O-CH₂-N₃；

Formulas I, formula J, formula K, formula L, formula M compound, R₅CHO and R₅CHO contracts in two fatty alcohols, R₅Selected from phenyl, 4- methoxybenzene Base, 4- chlorophenyl or 4- bromo phenyl；

In formula L ', formula M ' and formula N ' compound, R₈And R₉It is each independently selected from methyl, isopropyl or tert-butyl；

In formula E compound, X is chlorine；

In formula A- formula B compound, M is sodium.

4. method as claimed in claim 3, wherein

Formula C- formula O compound and R₁In OH, R₁Selected from methyl, ethyl, benzyl；

Formula C- formula O compound, formula J- formula O compound, acid anhydrides R₂-O-R₂, carboxylic acid halides R₇X and acid anhydrides R₇-O-R₇In, R₂And R₇Respectively It independently is acetyl group or benzoyl；

Formulas I, formula J, formula K, formula L, formula M compound, R₅CHO and R₅CHO contracts in two fatty alcohols, R₅For phenyl or 4- methoxybenzene Base；

In formula L ', formula M ' and formula N ' compound, R₈And R₉It is each independently methyl, isopropyl or tert-butyl；

In formula E compound, X is chlorine；

In formula A- formula B compound, M is sodium.

5. according to the method described in claim 1, wherein, in the step (1), the additional amount of hyaluronidase is formula A chemical combination The 2.0-3.0% of amount of substance, reaction temperature are 37 DEG C, and the molecular weight of formula A compound is 1KDa-10,000KDa；The faintly acid The pH value for referring to buffer solution is 3.0-6.5；The buffer solution is selected from acetic acid-sodium acetate buffer solution, acetic acid-potassium acetate buffering Liquid, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer or potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer one kind；The buffering is molten The concentration of liquid is 0.01M-1M；Concentration of the sodium chloride in weak acid buffer solution is 0.05M-0.5M.

6. according to the method described in claim 5, wherein, the molecular weight of formula A compound described in the step (1) is 10KDa- 1,000KDa；The buffer solution is the acetic acid-sodium acetate buffer solution that concentration is 0.1M；Concentration of the sodium chloride in buffer solution For 0.15M.

7. according to the method described in claim 1, wherein, in the step (2), the inorganic acid is selected from hydrogen chloride, sulfuric acid, nitre Acid, phosphoric acid；The alkaline condition is referred to selected from sodium acetate, pyridine, sodium dihydrogen phosphate, potassium carbonate, the alkali of triethylamine or piperidines Existing condition.

8. according to the method described in claim 7, wherein, in the step (2), the inorganic acid is selected from hydrogen chloride or sulfuric acid； The alkaline condition refers to condition existing for the alkali selected from pyridine or sodium acetate.

9. according to the method described in claim 1, wherein, in the step (3), the alkali is selected from ethylenediamine, hydrazine hydrate or 3- N, N dimethyl amino propylamine；The organic solvent is selected from acetone, chloroform, tetrahydrofuran, methylene chloride, N, N dimethyl formyl Amine or N, N dimethyl acetamide.

10. according to the method described in claim 9, wherein, in the step (3), the alkali is selected from hydrazine hydrate or 3-N, N diformazan Base amino propylamine；The organic solvent is selected from methylene chloride, tetrahydrofuran or N, N-dimethylformamide.

11. in the step (4), the alkali is selected from the carbonate of alkali metal, 1 according to the method described in claim 1, wherein, 11 carbon -7- alkene of 8- diazabicylo, one of acetate, borate, sodium hydroxide, potassium hydroxide；In three haloacetonitriles Halogen be selected from fluorine, chlorine, bromine.

12. according to the method for claim 11, wherein in the step (4), the alkali is selected from potassium carbonate, 1,8- phenodiazine One of miscellaneous two rings, 11 carbon -7- alkene；Halogen in three haloacetonitriles is chlorine.

13. according to the method described in claim 1, wherein, in the step (5), the acid is selected from trifluoromethanesulfonic acid alkyl silicon Rouge, boron trifluoride ether, copper chloride, p-methyl benzenesulfonic acid or camphorsulfonic acid.

14. according to the method described in claim 1, wherein, in the step (6), the acid is selected from trifluoromethanesulfonic acid alkyl silicon Rouge, boron trifluoride ether, copper chloride, p-methyl benzenesulfonic acid, camphorsulfonic acid；The organic solvent is selected from methylene chloride, chloroform, four One of hydrogen furans, acetone or 1,2- dichloroethanes.

15. according to the method for claim 14, wherein in the step (6), the acid copper chloride, trifluoromethanesulfonic acid C1- The silicone grease that C4 trialkyl replaces；The organic solvent is chloroform.

16. according to the method described in claim 1, wherein, in the step (7), the acid selected from hydrogen chloride, sulfuric acid, nitric acid, One of phosphoric acid or boron trifluoride ether；The alkali is selected from R₁ONa、R₁OK, NaOH, wherein R₁Selected from unsubstituted benzyl or Substituted benzyl, C1-C4 alkyl or allyl；Here, the substituted benzyl refers to that phenyl ring is replaced by one or more Replaced base, the substituent group is selected from halogen, nitro, C1-C4 alkyl, C1-C4 halogenated alkyl, C1-C4 alkoxy；Also, institute Substituent group is stated in any position of phenyl ring；

In the step (8), the acid is selected from p-methyl benzenesulfonic acid or camphorsulfonic acid；

In the step (9), the alkaline condition is referred to selected from sodium acetate, pyridine, sodium dihydrogen phosphate, potassium carbonate, triethylamine Or condition existing for the alkali of piperidines；

In the step (10), the acid is selected from one of trifluoroacetic acid, acetic acid, boric acid；The reducing agent is selected from trialkyl Substituted silane, sodium cyanoborohydride, borine；

In the step (11), the oxidant is selected from Dai Si-Martin's oxidant, the combination of acid anhydrides-dimethyl sulfoxide, oxalyl Combination, trifluoroacetic anhydride-dimethyl sulfoxide-triethylamine combination or the manganese dioxide of chloro- dimethyl sulfoxide-triethylamine；

In the step (12), the reducing agent is selected from sodium borohydride, lithium borohydride, potassium borohydride or alkyl-substituted hydroboration Salt；

In the step (13), the oxidant is selected from DDQ or ammonium ceric nitrate；The reducing agent is selected from triethylsilane and simple substance The combination of iodine or the combination of triethylsilane and carbon monoxide and cobalt octacarbonyl；

In the step (10a), the acid is selected from one of trifluoroacetic acid, acetic acid, boric acid；

In the step (11a), the substituted chlorosilane is selected from tert-butyl diphenyl chlorosilane, tert-butyldimethylsilyl chloride silicon Alkane, tri isopropyl chlorosilane, chlorotriethyl silane, 3,5-dimethylphenyl chlorosilane or trim,ethylchlorosilane；The alkali is selected from three second Amine, pyridine or 4-dimethylaminopyridine；

In the step (12a), the oxidant is selected from the combination of the chloro- dimethyl sulfoxide-triethylamine of oxalyl, acetic anhydride-diformazan Combination, manganese dioxide or the high iodine reagent of Dai Si-Martin of base sulfoxide；

In the step (13a), the reducing agent is selected from sodium borohydride, lithium borohydride, potassium borohydride or alkyl-substituted boron hydrogen Salt dissolving；

In the step (14), the acid is selected from p-methyl benzenesulfonic acid or trifluoroacetic acid；The fluorine ion reagent is selected from tetrabutyl fluorine Change ammonium, hydrogen fluoride pyridine complex compound, hydrogen fluoride triethylamine complex or ammonium fluoride.

17. according to the method for claim 16, wherein in the step (7), the acid is selected from hydrogen chloride or boron trifluoride Ether；The alkali is R₁ONa, wherein R₁Selected from methyl, ethyl, allyl, benzyl, 4- methoxy-benzyl；

In the step (9), the alkaline condition refers to condition existing for the alkali selected from pyridine or sodium acetate；

In the step (10), the acid is trifluoroacetic acid；The reducing agent is selected from the silane or cyano boron hydrogen that trialkyl replaces Change sodium；

In the step (11), the oxidant is Dai Si-Martin's oxidant；

In the step (12), the reducing agent is selected from triisobutyl potassium borohydride or sodium borohydride；

In the step (10a), the acid is acetic acid；

In the step (11a), the substituted chlorosilane is tert-butyl chloro-silicane；The alkali is pyridine；

In the step (12a), the oxidant is the high iodine reagent of Dai Si-Martin；

In the step (13a), the reducing agent is selected from triisobutyl potassium borohydride or sodium borohydride；

In the step (14), the acid is p-methyl benzenesulfonic acid；The fluorine ion reagent is hydrogen fluoride pyridine complex compound.

18. the midbody compound for synthesizing chondroitin sulfate tetrose, including following formula D- formula N ' compound:

Wherein, R₁Selected from unsubstituted benzyl or substituted benzyl, C1-C4 alkyl or allyl；Here, the substituted benzyl Refer to phenyl ring replaced one or more substituent group, the substituent group is selected from halogen, nitro, C1-C4 alkyl, C1-C4 Halogenated alkyl, C1-C4 alkoxy；Also, the substituent group is in any position of phenyl ring；

R₂And R₇It is each independently selected from aliphatic acyl radical, unsubstituted benzoyl or substituted benzoyl；Here, described Aliphatic acyl radical refers to C2-C6 alkanoyl；The substituted benzoyl refers to substituent group of the phenyl ring by one or more Replaced, the described substituent group is selected from halogen, nitro, C1-C4 alkyl, C1-C4 halogenated alkyl, C1-C4 alkoxy；Also, it is described Substituent group is in any position of phenyl ring；

R₃Selected from-(CH₂CH₂O)n-CH₂CH₂, n is 1-100 here；Aliphatic alkane subunit；Aliphatic olefin subunit；Or- (CH₂CH₂O)m-CH₂, m is 1-100 here；R₄Selected from azido, alkynyl, biotin, substituted amino, aldehyde radical, thioether；Institute It states substituted amino and refers to that the amino protected by protecting group, the protecting group are selected from benzyloxy carbonyl acyl group, tertiary butyloxycarbonyl acyl group, trichlorine Acetyl group, trifluoroacetyl group；

R₅Selected from unsubstituted phenyl or substituted phenyl, here, the substituted phenyl refers to phenyl ring by one or more Replaced substituent group, the substituent group is selected from halogen, nitro, C1-C4 alkyl, C1-C4 halogenated alkyl, C1-C4 alkoxy；And And the substituent group is in any position of phenyl ring；

R₈And R₉It is each independently selected from C1-C4 alkyl, phenyl or substituted phenyl, here, the substituted phenyl refers to benzene For ring replaced one or more substituent group, the substituent group is selected from halogen, nitro, C1-C4 alkyl, C1-C4 alkyl halide Base, C1-C4 alkoxy；Also, the substituent group is in any position of phenyl ring；

X is selected from fluorine or chlorine.

19. midbody compound as claimed in claim 18, wherein

R₁Selected from methyl, ethyl, allyl, benzyl, 4- methoxy-benzyl；

R₂And R₇It is each independently selected from acetyl group, benzoyl, 4- chlorobenzoyl base, 4- bromobenzoyl base；

-OR₃-R₄For-O-CH₂CH₂-O-CH₂-N₃；

R₅Selected from phenyl, 4- methoxyphenyl, 4- chlorophenyl or 4- bromo phenyl；

R₈And R₉It is each independently selected from methyl, isopropyl or tert-butyl；

In formula E compound, X is chlorine.

20. midbody compound as claimed in claim 19, wherein

R₁Selected from methyl, ethyl, benzyl；

R₂And R₇It is each independently acetyl group or benzoyl；

-OR₃-R₄For-O-CH₂CH₂-O-CH₂-N₃；

R₅For phenyl or 4- methoxyphenyl；

R₈And R₉It is each independently methyl, isopropyl or tert-butyl；

X is chlorine.

21. midbody compound as claimed in claim 18, comprising:

(bis--O- acetyl group -2- of 4,6- is de- for (tri--O- acetyl group of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) - Oxygen-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group of 2,3--beta d glucopyranosiduronic acid first Ester group) two-O- acetyl group -2- '-deoxy-n of-(1 → 3) -4,6--acetylaminohydroxyphenylarsonic acid D- glucopyranose；

O- (((tri--O- acetyl group of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group of 4,6-- 2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β-D- glucopyra alditol of 2,3- Sour carbomethoxy)-(1 → 3) -) two-O- acetyl group -2- '-deoxy-n of-O-4,6--acetylaminohydroxyphenylarsonic acid D- glucopyranosyl) three chloroethenes Imide ester；

(bis--O- acetyl group -2- of 4,6- is de- for (tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-O- Oxygen-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β of 2,3--D-Glucose aldehydic acid methyl esters Base)-(1 → 3) -2- methyl-(bis--O- acetyl group -1,2- dideoxy-α-D- glucopyranose of 4,6-) [2,1-d] 2- oxazoline；

1- nitrine -2- oxa- -4-O- [(tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid methyl esters)-(1 → 3)-(4,6- bis- - O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis- Portugal-O- acetyl group-β-D- 2,3- Grape uronic acid carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- glucopyranose of 4,6- Base)] butane；

1- nitrine -2- oxa- -4-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetyl ammonia Base-β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- second Acylamino--α-D- glucopyranosyl)] butane；

[(4,6-O- is to ar-methoxy-benzylidene -2- for (beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-by 1- nitrine -2- oxygen -4-O- Deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) - (4,6-O- is to benzylidene -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] butane；

[(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -2- oxa- -4-O- (4,6-O- is to ar-methoxy-benzylidene -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzene of 2,3- Formoxyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- deoxidation -2-N- acetyl ammonia Base-β-D- glucopyranosyl)] butane；

[(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -2- oxa- -4-O- (6-O- is to methoxybenzyl -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl of 2,3- Base-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- Glucopyranosyl)] butane；

[(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -2- oxa- -4-O- (6-O- is to methoxybenzyl -4- ketone -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- of 2,3- Benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -4- ketone -2- deoxidation -2-N- second Acylamino--β-D- glucopyranosyl)] butane；

[(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) -1- nitrine -2- oxa- -4-O- (6-O- is to methoxybenzyl -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(bis--O- benzoyl of 2,3- Base-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- Galactopyranosyl glycosyl)] butane；

1- nitrine -6-O- [(tri--O- acetyl group-β of 2,3,4--D-Glucose aldehydic acid methyl esters)-(1 → 3)-(bis--O- acetyl of 4,6- Base -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- acetyl group-β of 2,3--D-Glucose aldehyde Sour carbomethoxy)-(1 → 3)-(bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- glucopyranosyl of 4,6-)] hexane；

1- nitrine -6-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- pyrrole Glucopyranoside base)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α - D- glucopyranosyl)] hexane；

1- nitrine -6-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- benzal -2- deoxidation -2-N- second Acylamino--β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- benzal Base -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane；

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- Benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl-β-D- pyrrole of 2,3- Glucopyranoside aldehydic acid carbomethoxy)-(1 → 3)-(4,6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranose Base)] hexane；

[(2- is de- for (tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-by 1- nitrine -6-O- Oxygen -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzoyl-β-D- glucopyra alditol of 2,3- Sour carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane；

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- bis- Methyl tertbutyl silicon ether -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(bis--O- benzene first of 2,3- Acyl-beta-D- glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -2- deoxidation -2-N- acetyl Amino-beta-D- glucopyranosyl)] hexane；

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- bis- Methyl tertbutyl silicon ether -4- ketone -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(bis--O- of 2,3- Benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -4- ketone -2- is de- Oxygen -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane；

1- nitrine -6-O- [(tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- bis- Methyl tertbutyl silicon ether -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(bis--O- benzene first of 2,3- Acyl-beta-D- glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -2- deoxidation -2-N- acetyl Amino-beta-D- galactopyranosyl glycosyl)] hexane；With

[(2- is de- for (tri--O- benzoyl of 2,3,4--beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-by 1- nitrine -6-O- Oxygen -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(bis--O- benzoyl-β-D- glucopyra alditol of 2,3- Sour carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane.