CN106467563A

CN106467563A - The synthetic method of chondroitin sulfate tetrose and its midbody compound

Info

Publication number: CN106467563A
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: 2017-03-01
Anticipated expiration: 2036-08-19
Also published as: CN106467563B

Abstract

This application discloses a kind of synthetic method of chondroitin sulfate tetrose.With hyaluronate as initiation material; successively through enzymolysis; protection group is protected; protection group is changed; selectivity deprotection and hydroxyl oxidize obtain the midbody compound as shown in formula O with selective reduction; midbody compound synthesis chondroitin sulfate tetrose shown in recycling formula O, in formula O, being specifically defined of substituent group sees description.Additionally, additionally providing the midbody compound of synthesis chondroitin sulfate tetrose.The synthetic route that synthetic method provided herein relatively has document is easy, and without glycosylation, reaction yield is high, has a good application prospect.

Description

The synthetic method of chondroitin sulfate tetrose and its midbody compound

Technical field

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

Background technology

Chondroitin sulfate is a kind of glycosaminoglycans being widely present in vivo, and its structure is by glucuronic acid and N- acetylamino Galactose is constituted, and in its structure, hydroxyl there may be different degrees of sulphation, thus referred to as chondroitin sulfate.Chondroitin sulfate Element forms Dan Baiduotang proteoglycan PG with the albumen covalent bond in extracellular matrix, is distributed widely in intercellular substance, is a kind of particularly significant Glycosaminoglycans.Chondroitin sulfate take part in biochemical process much particularly important in vivo, for example：Take part in cell-ECM phase Interaction, the regeneration of suppression neural axon, the diffusion of impact cancerous cell and transfer, numerous cytokines combine with extracellular Deng.

In recent years it has been found that the different subtype of chondroitin sulfate, different sulphation modifications are had on specific hydroxyl Chondroitin sulfate A, C, E can mediate different physiologic functions.For example, chondroitin sulfate E can significance suppression Dorsal ganglion The growth of ganglion cell's aixs cylinder, and chondroitin sulfate A and C then no this effect.For the different sulfation sites of research and sulphation number Impact to chondroitin sulfate biological function, need to solve the chondroitin sulfate that these have specific sulphation pattern first can Obtain sex chromosome mosaicism.Traditional by extracting from natural origin, such as extract from the cartilage of animal, dissimilar sulfur can only be obtained The mixture of aching and limp ossein, further isolates and purifies and is difficult to.These extracts can meet a certain degree of medicine Learning research needs, for example, can be used for treatment of osteoarthritis etc..However, the mixture with different sulfation sites can not be expired Foot deeper medicineization research.Therefore, chemosynthesis are that current acquisition has clear and definite sulfation sites, and structure homogeneous can Carry out the only effective method of deeper biology and the chondroitin sulfate of medicineization research.

In prior art, synthesize chondroitin sulfate usually with glucuronic acid and N- acetylamino gala by chemical means Sugar or 2-Acetamido-2-deoxy-D-glucose monosaccharide are raw material, also have glycosylation to operate to carry out oligosaccharide by conventional protection group protection Assembling (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 many, and route is long, leads to the receipts of whole piece route due to being related to glycosylation operation Rate is low.At present, only synthesis chondroitin sulfate tetrose is accomplished by about 23 step reactions, and yield is less than 1%.Therefore, develop new sulfur The chemical synthesis process of aching and limp ossein oligosaccharide, significant for the medicineization research promoting chondroitin sulfate.

At present, it is both needed to be related to the post-synthetic phase in chondroitin sulfate to the exposed hydroxyl on the oligosaccharide backbone having been built up Base carries out sulfonated and finally entirely sugared deprotection.Therefore, how rapid build can be carried out in the middle of the oligosaccharide of sulphation modification Body is problem demanding prompt solution.An object of the application provides such a can directly carry out in the oligosaccharide of subsequent sulphuric acid modification The synthetic method of mesosome.

Content of the invention

Present inventor is to be easier to produce, to be easier to extract the hyaluronate sodium obtaining as raw material, by entering to raw material Row hydrolysis, it has been chemically modified to obtain and can be used for chondroitin sulfate A, the key intermediate of tri- kinds of tetroses synthesis of C, E, overcome existing The deficiency that technology exists.

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

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

In the embodiment of the application, there is provided a kind of synthetic method of chondroitin sulfate tetrose, methods described includes The synthesis of chondroitin sulfate tetrose key intermediate, comprises the 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 in the presence of enzyme；Here, described hyaluronidase derives from animal testis, vigor 400-1000IU/ mg；Preferably, selected from bull testis or sheep testicle；More preferably bull testis；

(2) formula B compound is in the alcohol R of mineral acid₁Carry out esterification in OH solution, the carboxyl of formula B compound is carried out Esterification；Then utilize anhydride R₂-O-R₂, in the basic conditions remaining all hydroxyl of formula B compound are carried out being acylated protection, obtain To formula C compound；

Here, anhydride 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 2-Acetamido-2-deoxy-D-glucose in organic solvent 1 hydroxyl, obtains formula D compound；

(4) formula D compound, under base catalysiss, is reacted with three haloacetonitrile, obtains the formula E compound that end group contains imines ester；

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

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

Here, alcohol R₄-R₃R in OH₃It is selected from-CH₂CH₂-(OCH₂CH₂) n-, n is 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, the amino replacing, aldehyde radical, thioether, described substituted amino refers to the amino protected by protection group, described Protection group is selected from benzyloxy carbonyl acyl group, tertiary butyloxycarbonyl acyl group, tribromo-acetyl base, trifluoroacetyl group；Preferably, R₄-R₃OH is N₃- CH₂-OCH₂CH₂-OH；

(7) formula G compound is in the alcohol R of acid or alkali₁In OH solution, slough all of R on formula G compound₂, obtain formula H chemical combination Thing；

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

Here, R₅CHO or R₅CHO contracting two fatty alcohol (such as R₅CH(OCH₃)₂) in R₅Selected from phenyl or substituted phenyl；

(9) compound of formula I is in carboxylic acid halides R₇X or anhydride R₇-O-R₇In the presence of, it is acylated in the basic conditions, obtained formula J Compound；

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

(10) under the existence condition of acid and reducing agent, exposed 4 hydroxyls of selectivity are so that 6 is R to formula J compound₅- CH₂- protection, obtains formula K compound；

(11) formula K compound, under oxidant existence condition, is aoxidized to 4,4 ' position hydroxyls, is obtained formula L compound；

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

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

Or, step (10)-(13) are replaced with following step (10a)-(14)：

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

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

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

(13a) formula M ' compound under reducing agent existence condition by the ketone carbonyl of 4 and 4 ' positions of 2-Acetamido-2-deoxy-D-glucose Stereoselective be reduced to axial bond hydroxyl, obtain formula N ' compound；

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

(15) utilize formula O compound synthesis chondroitin sulfate tetrose；

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 Pi-allyl；Here, described substituted benzyl refers to that phenyl ring is replaced by one or more substituent group, and described substituent group is selected from Halogen (such as fluorine, chlorine, bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl Base or the tert-butyl group), C1-C4 haloalkyl (such as trifluoromethyl), C1-C4 alkoxyl (such as methoxyl group, ethyoxyl, positive third oxygen Base, isopropoxy, n-butoxy, isobutoxy or tert-butoxy)；And, described substituent group is in the optional position of phenyl ring；Preferably Ground, R₁Selected from methyl, ethyl, pi-allyl, benzyl, 4- methoxy-benzyl, it 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, described aliphatic acyl radical refers to C2-C6 alkanoyl (such as acetyl group, propionyl Base, positive bytyry, isobutyryl, positive valeryl, valeryl or positive caproyl), described substituted benzoyl refers to phenyl ring Replaced by one or more substituent group, described substituent group is selected from halogen (such as fluorine, chlorine, bromine or iodine), nitro, C1- C4 alkyl (such as methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group or the tert-butyl group), C1-C4 haloalkyl are (for example Trifluoromethyl), C1-C4 alkoxyl (for example methoxyl group, ethyoxyl, positive propoxy, isopropoxy, n-butoxy, isobutoxy or Tert-butoxy)；And, described substituent group is in the optional 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, selected from-CH₂CH₂-O-CH₂-；R₄It is selected from Azido, alkynyl, biotin, the amino replacing, aldehyde radical, thioether；Amino that described substituted amino refers to be protected by protection group, Described protection group is selected from benzyloxy carbonyl acyl group, tertiary butyloxycarbonyl acyl group, tribromo-acetyl base, trifluoroacetyl group；Preferably ,-OR₃-R₄For- O-CH₂CH₂-O-CH₂-N₃；

In Formulas I, formula J, formula K, formula L and formula M compound, R₅Selected from unsubstituted phenyl or substituted phenyl, here, described Replace phenyl refer to that phenyl ring is replaced by one or more substituent group, described substituent group be selected from halogen (for example fluorine, chlorine, Bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group or the tert-butyl group), C1- C4 haloalkyl (such as trifluoromethyl), C1-C4 alkoxyl (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, positive fourth Epoxide, isobutoxy or tert-butoxy)；And, described substituent group is in the optional position of phenyl ring；Preferably, selected from phenyl, 4- first Phenyl, 4- chlorophenyl, 4- bromo phenyl, it is highly preferred that be selected from phenyl or 4- methoxyphenyl；

Formula L ', formula M ' and formula N ' in compound, R₈And R₉It is each independently selected from C1-C4 alkyl, phenyl or substituted benzene Base；Here, described C1-C4 alkyl is selected from methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group or the tert-butyl group；Described Replace phenyl refer to that phenyl ring is replaced by one or more substituent group, described substituent group be selected from halogen (for example fluorine, chlorine, Bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group or the tert-butyl group), C1- C4 haloalkyl (such as trifluoromethyl), C1-C4 alkoxyl (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, positive fourth Epoxide, isobutoxy or tert-butoxy)；And, described substituent group is in the optional position of phenyl ring；Preferably, R₈And R₉Each independent Ground is selected from methyl, isopropyl or the tert-butyl group 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 is it is therefore preferable to sodium.

The application provide chondroitin sulfate tetrose synthetic method in, in described step (1), hyaluronidase plus Enter the 2.0-3.0% that amount is hyaluronate sodium quality, reaction temperature is 37 DEG C, the molecular weight of formula A compound is 1KDa-10, 000KDa is it is therefore preferable to 10KDa-1,000KDa；Described faintly acid refer to buffer solution pH value be 3.0-6.5 it is therefore preferable to 5.0-5.2；Described buffer solution is selected from acetic acid-sodium acetate buffer solution, acetic acid -50 mM, sodium dihydrogen phosphate-phosphoric acid hydrogen One kind of disodium buffer or potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer is it is therefore preferable to from acetic acid-sodium acetate buffer solution；Institute The concentration stating buffer elects 0.01M-1M as；Preferably, described buffer solution is the acetic acid-acetate buffer of 0.1M for concentration Liquid；Concentration in weak acid buffer solution for the sodium chloride is 0.05M-0.5M it is therefore preferable to 0.15M；Described hyaluronidase choosing From the hyaluronidase of animal testicular origin, the hyaluronic acid in the bull testis source for example bought from Sigma-Aldrich company Enzyme.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (2), the choosing of described mineral acid From hydrogen chloride, sulphuric acid, nitric acid, phosphoric acid it is preferable that being selected from hydrogen chloride or sulphuric acid；Alcohol R₁R in OH₁As mentioned above；Described alkalescence bar Part refers to selected from Sodium Acetate Trihydrate, pyridine, sodium dihydrogen phosphate, potassium carbonate, the condition that the alkali of triethylamine or piperidines exists it is preferable that Refer to the condition existing selected from the alkali of pyridine or Sodium Acetate Trihydrate.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (3), described alkali is selected from second two Amine, hydrazine hydrate or 3-N, N dimethyl amino propylamine is it is preferable that be selected from hydrazine hydrate or 3-N, N dimethyl amino propylamine；Described have Machine solvent selected from acetone, chloroform, oxolane, dichloromethane, DMF or N, N dimethyl acetamide, preferably Ground, selected from dichloromethane, oxolane or DMF.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (4), described alkali is selected from alkali gold The carbonate of genus, 1,8- diazabicylo 11 carbon -7- alkene (DBU), in acetate, borate, sodium hydroxide, potassium hydroxide A kind of；Preferably, selected from one of potassium carbonate or 1,8- diazabicylo 11 carbon -7- alkene (DBU)；Described three haloacetonitrile In halogen be selected from fluorine, chlorine, bromine it is therefore preferable to chlorine.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (5), described acid is selected from trifluoro Methanesulfonic acid alkyl silicone grease, boron trifluoride diethyl etherate, copper chloride, p-methyl benzenesulfonic acid, or camphorsulfonic acid.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (6), described acid is selected from trifluoro Methanesulfonic acid alkyl silicone grease, boron trifluoride diethyl etherate, copper chloride, p-methyl benzenesulfonic acid, camphorsulfonic acid；Preferably, selected from copper chloride, three The silicone grease that fluorine methanesulfonic acid C1-C4 trialkyl replaces；Described organic solvent is selected from dichloromethane, chloroform, oxolane, acetone or 1, One of 2- dichloroethanes；It is preferably chloroform.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (7), described acid is selected from chlorination One of hydrogen, sulphuric acid, nitric acid, phosphoric acid or boron trifluoride diethyl etherate are it is preferable that be selected from hydrogen chloride or boron trifluoride diethyl etherate；Described Alkali is selected from R₁ONa、R₁OK or NaOH is it is therefore preferable to R₁ONa, wherein, R₁Selected from unsubstituted benzyl or substituted benzyl, C1-C4 Alkyl or pi-allyl；Here, described substituted benzyl refers to that phenyl ring is replaced by one or more substituent group, described takes Dai Ji is selected from halogen (such as fluorine, chlorine, bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-pro-pyl, isopropyl, just Butyl, isobutyl group or the tert-butyl group), C1-C4 haloalkyl (such as trifluoromethyl), C1-C4 alkoxyl (such as methoxyl group, ethoxy Base, positive propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy)；And, described substituent group is any in phenyl ring Position；Preferably, R₁Selected from methyl, ethyl, pi-allyl, benzyl, 4- methoxy-benzyl, it is more preferably selected from methyl, ethyl, benzyl Base.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (8), described acid is selected to first Base benzenesulfonic acid or camphorsulfonic acid.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (9), described alkalescence condition refers to Be selected from Sodium Acetate Trihydrate, pyridine, sodium dihydrogen phosphate, potassium carbonate, triethylamine or piperidines alkali exist condition it is preferable that referring to It is the condition of the alkali presence selected from pyridine or Sodium Acetate Trihydrate.

In the synthetic method of the chondroitin sulfate that the application provides, in described step (10), described acid is selected from trifluoro second One of acid, acetic acid, boric acid are it is therefore preferable to trifluoroacetic acid；Described 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；Preferably Ground, silane (such as triethyl silicane, tri isopropyl silane, t-butyldimethyl silane, the phenyl diformazan replacing selected from trialkyl Base silane) or sodium cyanoborohydride.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (11), described oxidant choosing From Dai Si-Martin's oxidant, the combination of anhydride-dimethyl sulfoxide, the combination of the chloro- dimethyl sulfoxide-triethylamine of oxalyl, trifluoro second The combination of anhydride-dimethyl sulfoxide-triethylamine or manganese dioxide；Preferably, it is Dai Si-Martin's oxidant；

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (12), described reducing agent choosing From sodium borohydride, lithium borohydride, potassium borohydride, alkyl replace borohydride salts (for example：Triisobutyl potassium borohydride or three isobutyls Base lithium borohydride)；Preferably, selected from triisobutyl potassium borohydride or sodium borohydride.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (13), described oxidant choosing From DDQ or ammonium ceric nitrate；Described reducing agent is selected from combining or triethyl silicane and carbon monoxide of triethyl silicane and iodine With combining of cobalt octacarbonyl.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (10a), described acid is selected from three One of Fluoroethanoic acid, acetic acid, boric acid are it is preferable that be acetic acid.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (11a), described 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；Preferably, it is tert-butyl chloro-silicane；Described alkali is selected from Triethylamine, pyridine or DMAP are it is preferable that be pyridine.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (12a), described 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；It is preferably the high iodine reagent of Dai Si-Martin.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (13a), described reducing agent choosing From sodium borohydride, lithium borohydride, potassium borohydride or alkyl replace borohydride salts (for example：Triisobutyl potassium borohydride or three different Butyl lithium borohydride)；Preferably, selected from triisobutyl potassium borohydride or sodium borohydride.

In the synthetic method of the chondroitin sulfate tetrose that the application provides, in described step (14), described acid is selected from right Toluenesulfonic acid or trifluoroacetic acid, preferably p-methyl benzenesulfonic acid；Described fluorion reagent is selected from tetrabutyl ammonium fluoride, hydrogen fluoride pyridine Complex, fluohydric acid gas triethylamine complex or ammonium fluoride；It is preferably hydrogen fluoride pyridine complex.

The application provide chondroitin sulfate tetrose synthetic method, 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 complex of 20-100 equivalent to hydroxyl thereon Base carries out sulfonic acid esterification, then sloughs protection group in the basic conditions and obtains chondroitin sulfate E tetrose, i.e. formula CS-E compound；

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

(iii) formula O compound and Cyanophenacyl in the presence 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 complex to 4 with The hydroxyl of 4 ' positions carries out sulfonic acid esterification, then sloughs protection group in the basic conditions and obtain chondroitin sulfate A tetrose, i.e. formula CS-Aization Compound；

Here, step (i), the sulfur trioxide organic base complex in (ii) and (iii) are selected from sulfur trioxide pyridine and are combined Thing, sulfur trioxide triethylamine complex or sulfur trioxide trimethylamine complex are it is preferable that be sulfur trioxide trimethylamine complex. Described alkalescence condition refers to the condition that following alkali exists：Sodium hydroxide or its mixed solution with hydrogen peroxide, potassium hydroxide Or its mixed solution with hydrogen peroxide, Lithium hydrate or its mixed solution with hydrogen peroxide, it is preferably chosen from Lithium hydrate Or its mixed solution with hydrogen peroxide.

Additionally, in the synthetic method of the chondroitin sulfate tetrose that the application provides, after obtaining formula O compound, can join Examine the teaching (list of references of prior art：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.) prepare chondroitin sulfate tetrose A, C or E.

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

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

Formula D- formula N ' in compound, R₁Selected from unsubstituted benzyl or substituted benzyl, C1-C4 alkyl or pi-allyl；This In, described substituted benzyl refers to that phenyl ring is replaced by one or more substituent group, and described substituent group is selected from halogen (for example Fluorine, chlorine, bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group or tertiary fourth Base), C1-C4 haloalkyl (such as trifluoromethyl), C1-C4 alkoxyl (such as methoxyl group, ethyoxyl, positive propoxy, isopropyl oxygen Base, n-butoxy, isobutoxy or tert-butoxy)；And, optionally, described substituent group is in the optional position of phenyl ring；Preferably Ground, R₁Selected from methyl, ethyl, pi-allyl, benzyl, 4- methoxy-benzyl, it is more preferably selected from methyl, ethyl, benzyl；

Formula D- formula N ' in compound, R₂And R₇It is each independently selected from aliphatic acyl radical, unsubstituted benzoyl or replacement Benzoyl；Here, described aliphatic acyl radical refers to C2-C6 alkanoyl (such as acetyl group, propiono, positive bytyry, isobutyl Acyl group, positive valeryl, valeryl or positive caproyl)；Described substituted benzoyl refers to phenyl ring by one or more Substituent group replaced, described substituent group be selected from halogen (such as fluorine, chlorine, bromine or iodine), nitro, C1-C4 alkyl (such as methyl, Ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group or the tert-butyl group), C1-C4 haloalkyl (such as trifluoromethyl), C1-C4 alkane Epoxide (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy)；And, appoint Selection of land, described substituent group is in the optional 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；

Formula G- formula N ' in 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, selected from-CH₂CH₂-O-CH₂-；R₄ Selected from azido, alkynyl, biotin, the amino replacing, aldehyde radical, thioether；Described substituted amino refers to be protected by protection group Amino, described protection group is selected from benzyloxy carbonyl acyl group, tertiary butyloxycarbonyl acyl group, tribromo-acetyl base, trifluoroacetyl group；Preferably, OR₃-R₄ For-O-CH₂CH₂-O-CH₂-N₃；

In Formulas I, formula J, formula K, formula L and formula M compound, R₅Selected from unsubstituted phenyl or substituted phenyl, here, described Replace phenyl refer to that phenyl ring is replaced by one or more substituent group, described substituent group be selected from halogen (for example fluorine, chlorine, Bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group or the tert-butyl group), C1- C4 haloalkyl (such as trifluoromethyl), C1-C4 alkoxyl (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, positive fourth Epoxide, isobutoxy or tert-butoxy)；And, optionally, described substituent group is in the optional position of phenyl ring；Preferably, selected from benzene Base, 4- methoxyphenyl, 4- chlorophenyl, 4- bromo phenyl, it is highly preferred that be selected from phenyl, 4- methoxyphenyl；

Formula L ', formula M ' and formula N ' in compound, R₈And R₉It is each independently selected from C1-C4 alkyl, phenyl or substituted benzene Base；Here, described C1-C4 alkyl refers to methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group or the tert-butyl group；Described Replace phenyl refer to that phenyl ring is replaced by one or more substituent group, described substituent group be selected from halogen (for example fluorine, chlorine, Bromine or iodine), nitro, C1-C4 alkyl (such as methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group or the tert-butyl group), C1- C4 haloalkyl (such as trifluoromethyl), C1-C4 alkoxyl (such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, positive fourth Epoxide, isobutoxy or tert-butoxy)；And, optionally, described substituent group is in the optional position of phenyl ring；Preferably, R₈And R₉ It is each independently selected from methyl, isopropyl, the tert-butyl group.

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

The midbody compound for synthesizing chondroitin sulfate tetrose that the application provides, including：

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

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

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

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

[(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to methoxy benzal for 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- [(2,3,4- tri--O- benzoyl-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- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -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- is to methoxybenzyl -2- deoxidation -2-N- acetyl ammonia Base-β-D- glucopyranosyl)] butane；

1- nitrine -2- oxa- -4-O- [(2,3,4- tri--O- benzoyl-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- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- is to methoxybenzyl -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(2,3- bis--O- 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- [(2,3,4- tri--O- acetyl group-β-D-Glucose aldehydic acid methyl ester)-(1 → 3)-(4,6- bis--O- Acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D- Fructus Vitis viniferae Alduronic acid carbomethoxy)-(1 → 3)-(4,6- bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- glucopyranosyl)] 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- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4, 6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- benzoyl-β - 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- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- Deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- benzoyl-β-D- Glucopyranose. Aldehydic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)] hexane；

1- nitrine -6-O- [(2,3,4- tri--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)-(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；

1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-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- [(2,3,4- tri--O- 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)-(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- galactopyranosyl glycosyl)] hexane；With

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

The application be easier to produce, be easier to extract the hyaluronate sodium obtaining as raw material, by cheap animal testis Lai The hyaluronidase in source is hydrolyzed to raw material, obtains the tetrose skeleton that can be used for being chemically modified afterwards, then it is entered The conventional protection group operation of row is thus synthesis can be used for chondroitin sulfate A, the key intermediate of tri- kinds of tetrose synthesis of C, E, whole piece Route need not carry out glycosylation operation, and reaction scheme is short, high income, and can have azido, is easy to later-stage utilization click reaction It is marked, be produced on a large scale chondroitin sulfate and its derivant；On the other hand, by suitable condition optimizing and purification side Formula selects, and can obtain the higher sample of purity and be used for studying, and the medicineization being easy to chondroitin sulfate is explored.

Specific embodiment

The exploitativeness of the application to be further described, not the limit to the application protection domain below by embodiment System.

Detecting instrument：

Nuclear-magnetism：Bruker AV-400 type nuclear magnetic resonance analyser, solvent is CDCl₃,CD₃OD, TMS are internal standard.

Mass spectrum：Bruker APEX IV type mass spectrograph.

Abbreviation：

CH₂Cl₂For dichloromethane

EtOAc is ethyl acetate

MeOH is methanol

DMF is N,N-dimethylformamide

DMP:Dai Si-Martin reagent

HRMS:High resolution mass spectrum

ESI:Electrospray Mass Spectrometry

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 the hyaluronic acid fully swelling at room temperature (about needing 72h) be with vigorous stirring Colourless, homogeneous sticky colloidal liquid.Make this colloidal liquid be warming up to 37.0 DEG C using water bath with thermostatic control, now add 1.0g transparent Matter acid enzyme dry powder (buying from Sigma-Aldrich company, article No. H-3506, Lot#SLBL1922V), and stir at 37.0 DEG C Two weeks.Reaction terminates for reactant liquor to be warming up to boiling, and stirs 15min at 105 DEG C, does not continue to after muddiness, it is rapid after solution It is cooled to room temperature, and vacuum distillation after mixing with the dehydrated alcohol of 1/20 volume, it is concentrated to dryness to obtain white syrup.Afterwards by this sugar Slurry is distilled water dissolution and is slowly dropped into precipitation white precipitate in the absolute ethanol that 3000mL is stirred vigorously with a small amount of, with filtration under diminished pressure Method be less than in 50% environment in relative air humidity and collect separated out precipitation, and washed with absolute ethanol cold on a small quantity Wash filter cake, then to constant weight, obtain collecting the pressed powder obtaining drying under reduced pressure in the exsiccator fill anhydrous calcium chloride 96.0g white amorphous powder shape solid.This crude product can be directly used for lower step synthesis, need not be further purified.Anti- for determining The product structure answered and yield, 200mg crude product is pure through LH-20 sephadex column (pillar height × column diameter=100cm × 2cm) Change, to distill water elution, obtain title tetrose product 60mg, with document P.Litant et al.Carbohydr.Res.237 (1992) 271-281 is provided¹H NMR spectra is more identical.

Embodiment 2：(2,3,4- tri--O- acetyl group-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6- bis-- O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D- pyrans Glucuronic acid methyl ester base)-(1 → 3) -1,4,6- three-O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid D- Glucopyranose. conjunction Become

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 new refrigeration of 0.08M.Then reactant liquor is stood 96h in 4 DEG C.Reaction knot After bundle, reactant liquor triethylamine is neutralized to neutrality, then vacuum distillation, and is concentrated to dryness.Add pyridine 405mL, be cooled to 0 DEG C, after being slowly added dropwise acetic anhydride 230mL, reactant liquor is slowly increased to room temperature, and stirs 24h at room temperature.After question response terminates, Reactant liquor is cooled down in ice-water bath, is slowly added dropwise methanol 170mL, and stir 1h at a temperature of this, by reactant liquor vacuum distillation It is concentrated to give brown color syrup.This syrup is dissolved in 2.5L EtOAc, filtration under diminished pressure removes insoluble matter, organic faciess are used successively The 1M aqueous hydrochloric acid solution of 400mL washs 3 times, and aqueous phase is extracted three times with 400mL EtOAc.Merge organic faciess, and use 600mL successively Saturated sodium bicarbonate aqueous solution washs 2 times, 600mL saturated common salt water washing 1 time.By organic faciess anhydrous sodium sulfate drying, mistake Filter, vacuum distillation, gained crude product dichloromethane:Ethyl acetate:Methanol=1:1:0.06 to 1:1:0.14 (eluant contains 0.1% triethylamine) Flash silica column chromatography purification, obtain 32.6g white, amorphous solid.Hyaluronate sodium 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 α：β=3:1 mixture.

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

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

Take (2,3,4- tri--O- acetyl group-β-D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(4,6- bis--O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- Glucopyranose .)-(1 → 4)-(2,3- bis--O- acetyl group-β-D-Glucose aldehydic acid methyl ester Base)-(1 → 3) -1,4,6- three-O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid D- Glucopyranose. 30.2g (24.6mmol) is molten In 240mL oxolane, it is slowly added dropwise 15.5mL (90.7mmol) 3-N under agitation, N- dimethyl-amino propylamine, and in 3h is stirred under room temperature.Reactant liquor is diluted with 650mL chloroform, organic faciess are washed 3 times with the 1M hydrochloric acid solution of 150mL, aqueous phase is used 150mL chloroform extraction 3 times, merges organic faciess, is washed with 230mL saturated sodium bicarbonate aqueous solution 1 time, saturated common salt water washing 1 Secondary, organic faciess anhydrous sodium sulfate drying, filters, concentrating under reduced pressure obtains the faint yellow amorphous powder of 25.5g, yield 88%.Thick product Thing need not be further purified, you can for following reaction.

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

Take dry (2,3,4- tri--O- acetyl group-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6- bis-- O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D- pyrans Glucuronic acid methyl ester base)-(1 → 3) -4,6- two-O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid D- Glucopyranose. crude product 7.0g (6.0mmol) is dissolved in 60mL dry methylene chloride, adds Tritox 6.7mL (60mmol), by this mixture in argon Cool down in ice-water bath under gas atmosphere.It is slowly added dropwise 1,8- diazabicylo 11 carbon -7- alkene (DBU) 149 μ L afterwards (0.9mmol).Reactant liquor stirs 2.5h at 0 DEG C.By reactant liquor concentrating under reduced pressure after the completion of question response, and with silica gel quick post color Spectrum purification (dichloromethane：Ethyl acetate：Methanol=3：1：0.10, containing 0.1% triethylamine) obtain 5.5g white, amorphous solid, receive 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]⁺Value of calculation 1326.2768, measured value [M+H]⁺：1326.2738.

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

Take dry O- (((2,3,4- tri--O- acetyl group-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6- Two-O- acetyl group -2- '-deoxy-ns-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D- Glucopyranosiduronic acid carbomethoxy)-(1 → 3) -)-O-4,6- two-O- acetyl group -2- '-deoxy-n-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 gas atmosphere Middle cooling.It is slowly added dropwise Trimethylsilyl trifluoromethanesulfonate 182 μ L (1.02mmol).Reactant liquor stirs 0.5h at 0 DEG C.Treat anti- After the completion of answering, Deca 150 μ L triethylamine is quenched reaction.Reactant liquor is evaporated to dry, and uses silica gel flash column chromatography (dichloromethane：Ethyl acetate：Methanol=3：1：0.15, containing 0.1% triethylamine) obtain 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):Value of calculation [M+H]⁺:1165.3566, measured value [M+H]⁺：1165.3561.

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

Take dry O- (2,3,4- tri--O- acetyl group-β-D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(4,6- bis--O- Acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D-Glucose Aldehydic acid carbomethoxy)-(1 → 3) -2- methyl-(4,6- bis--O- acetyl group -1,2- dideoxy-α-D- Glucopyranose .) [2,1-d] 2- oxazoline 5.0g (4.3mmol) is dissolved in the chloroform that 35mL is dried, and adds (its preparation of 1- nitrine -2- oxa- -4 butanol 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).Reactant liquor Add 10mL distilled water after backflow 10h at 65 DEG C and continue the 1h that flows back.Reactant liquor is cooled down at room temperature, adds unsaturated carbonate Hydrogen sodium water solution is simultaneously stirred vigorously.Aqueous phase is extracted 3 times with 10mL dichloromethane, merges organic faciess, is washed with saturation NaCl aqueous solution Wash, anhydrous sodium sulfate drying, filter, be concentrated to give colourless syrup.This syrup is scattered in 30mL absolute ether and is stirred vigorously 12h, it is seen that a large amount of white solid separates out, is collected by filtration solid, by solid silica gel flash column chromatography (dichloromethane：Second Acetoacetic ester：Methanol=1:1:0.10, containing 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):Value of calculation [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)-(1 → 4)-(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(2- takes off Oxygen -2-N- acetylaminohydroxyphenylarsonic acid α-D- glucopyranosyl)] butane

Take 1- nitrine -2- oxa- -4-O- [(2,3,4- tri--O- acetyl group-β-D-Glucose aldehydic acid methyl ester)-(1 → 3) - (4,6- bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl Base-β-D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(4,6- bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- pyrans Glucosyl group)] butane 4.0g (3.1mmol) is dissolved in the hydrogen chloride methanol solution of 30mL 0.25M.Reactant liquor stirs at room temperature Mix 36h, after the completion of question response, use " Amberlite " IRA-410OH^-Type anion exchange resin neutralization reaction liquid is to neutrality.Cross Filter off and remove resin, resin methanol washs 3 times, cleaning mixture is merged with filtrate and is evaporated to dry, and is filling anhydrous calcium chloride Exsiccator in be dried, obtain 2.8g white, amorphous solid, yield 100%.Product need not be further purified, you can for follow-up 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 ester 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 15mL and is dried in DMF, Add (+)-camphorsulfonic acid (0.22mmol) and P-methoxybenzal-dehyde dimethyl acetal (22mmol), reactant liquor is at reduced pressure conditions (0.5 atmospheric pressure) stirs 4h in 50 DEG C.Add triethylamine that reaction is quenched, reactant liquor evaporated under reduced pressure is obtained yellow syrup.Should Syrup is scattered in 20mL absolute ether and is stirred vigorously 12h it is seen that a large amount of white solid separates out, and solid is collected by filtration, solid is used The cold diethyl ether solution washing containing 20% methanol, obtains 1.9g crude product, yield 75% on a small quantity.This crude product need not be further purified, that is, Can be used for following reaction.

R_f=0.40 (SiO₂,CH₂Cl₂:MeOH=15:1)；

Embodiment 9：1- nitrine -2- oxa- -4-O- [(2,3,4- tri--O- benzoyl-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)-(2,3- bis--O- benzoyl-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

[(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to methoxy to take 1- nitrine -2- oxa- -4-O- Benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(beta d glucopyranosiduronic acid methyl ester 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, Benzenecarbonyl chloride. is previously dissolved in 1mL pyridine, is then slowly added dropwise in reactant liquor simultaneously Stir 3h in 0 DEG C.Reaction is quenched with saturated sodium bicarbonate aqueous solution, by reactant liquor evaporated under reduced pressure, with diluted ethyl acetate, organic Mutually washed with the aqueous hydrochloric acid solution of 0.5N successively, saturated sodium bicarbonate aqueous solution washs, saturation NaCl solution washing, anhydrous sulfur Sour sodium is dried, and filters, and concentrates.Concentrate precipitates to obtain 2.3g white, amorphous solid, yield through ether：85%.

R_f=0.63 (SiO₂,CH₂Cl₂:MeOH=25:1).

HRMS:Value of calculation [M+Na]⁺:1696.5277, measured value [M+Na]⁺：1696.5290.

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

Take dry O- (2,3,4- tri--O- acetyl group-β-D-Glucose aldehydic acid carbomethoxy)-(1 → 3)-(4,6- bis--O- Acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D-Glucose Aldehydic acid carbomethoxy)-(1 → 3) -2- methyl-(4,6- bis--O- acetyl group -1,2- dideoxy-α-D- Glucopyranose .) [2,1-d] 2- oxazoline 10.0g (8.6mmol) is dissolved in the chloroform that 90mL is dried, and (its synthetic method is joined to add 6- nitrine -1- hexanol 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).Reactant liquor is cooled to room temperature under an argon atmosphere after 65 DEG C of backflow 10h, adds unsaturated carbonate hydrogen Sodium water solution is simultaneously stirred vigorously.Aqueous phase is extracted 3 times with 10mL dichloromethane, merges organic faciess, with saturation NaCl solution washing, Anhydrous sodium sulfate drying, filters, is concentrated to give colourless syrup.This syrup is scattered in 60mL absolute ether and is stirred vigorously 12h, can See that a large amount of white solids separate out, filtration under diminished pressure collects separated out pressed powder, filter cake is washed with absolute ether, obtain in vain after being dried 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]⁺:Value of calculation 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- [(2,3,4- tri--O- acetyl group-β-D-Glucose aldehydic acid methyl ester)-(1 → 3)-(4,6- bis- - O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D- Portugal Grape alduronic acid carbomethoxy)-(1 → 3)-(4,6- bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- Glucopyranose. Base)] hexane 4.0g (3.1mmol) is dissolved in the hydrogen chloride methanol solution of 30mL 0.25M.Reactant liquor stirs 36h at room temperature, " Amberlite " IRA-410OH is used after the completion of question response^-Type anion exchange resin neutralization reaction liquid is to neutrality.Filter and remove Resin, resin methanol washs 3 times, and cleaning mixture is merged with filtrate and is evaporated to dry, and is filling the drying of anhydrous calcium chloride It is dried in device, obtain 2.8g white, amorphous solid, yield 100%.Product need not be further purified, you can for subsequent reactions.

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 68mL and is dried in DMF, add (+)- Camphorsulfonic acid 1.27g (5.5mmol) and P-methoxybenzal-dehyde dimethyl acetal 18.6mL (124mmol), reactant liquor is in decompression bar Under part, (0.5 atmospheric pressure) is stirred overnight in 50 DEG C.Add triethylamine that reaction is quenched, reactant liquor evaporated under reduced pressure is obtained yellow sugar Slurry.This syrup is scattered in 10mLCH₂Cl₂In, after be slowly added to 120mL absolute ether with vigorous stirring it is seen that in a large number white Solid separates out, and filtration under diminished pressure collects solid, and solid is washed with methanol cold on a small quantity with water successively, obtains 7.12g crude product, yield 84%.This crude product need not be further purified, you can for following reaction.

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]⁺:Value of calculation C₅₀H₆₈N₅O₂₃, 1106.4300, measured value 1106.4318；[M+Na]⁺:Meter Calculation value C₅₀H₆₇N₅NaO₂₃, 1128.4119, measured value 1128.4127；[M+K]⁺:Value of calculation C₅₀H₆₇N₅KO₂₃, 1144.3858, real Measured value 1144.3830.

Embodiment 13：1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-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

Take 1- nitrine -6-O- [(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- 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) it is dissolved in 60mL pyrido and is cooled to -15 DEG C, Benzenecarbonyl chloride. 2.8mL (23.9mmol) is slowly added dropwise into reactant liquor In and in 0 DEG C stir 3h.Reaction is quenched with saturated sodium bicarbonate aqueous solution, by reactant liquor evaporated under reduced pressure, with diluted ethyl acetate, Organic faciess are washed with the aqueous hydrochloric acid solution of 0.5N successively, and saturated sodium bicarbonate aqueous solution washs, saturation NaCl solution washing, no Aqueous sodium persulfate is dried, and filters, and concentrates.Concentrate precipitates to obtain 6.3g white, amorphous solid, yield through ether：86%.

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]⁺:Value of calculation C₈₅H₈₈N₅O₂₈, 1626.5610, measured value 1626.5626.

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

Take 1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) - (4,6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- benzoyl 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 will react after stirring 1h at a temperature of this Liquid is cooled to room temperature.With toluene together vacuum distillation 3 times.Residue is washed with absolute ether, filtration under diminished pressure, collects filter cake, does 6.0g white, amorphous solid, yield is obtained after dry：85%.

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]⁺:Value of calculation 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- [(2,3,4- tri--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)-(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

Take 1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) - (2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- benzoyl-β-D- glucopyra Alduronic 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 reactant liquor is stirred at room temperature overnight under an argon atmosphere.Treat thin layer After analysis Chromatogram display reaction completely, reactant liquor is cooled down in ice-water bath, Deca methanol is quenched reaction.Reactant liquor is reduced pressure dense Contracting, residue with Ethyl acetate dilutes, and uses 1M aqueous hydrochloric acid solution, saturated sodium bicarbonate aqueous solution, saturated common salt water washing successively. Organic faciess anhydrous sodium sulfate drying, filters, and concentrates, and 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]⁺:Value of calculation C₈₃H₁₀₈N₅O₂₈Si₂, 1678.6714, measured value 1678.6676.

Embodiment 16：1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-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)-(2,3- bis--O- benzoyl-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 1- nitrine -6-O- [(2,3,4- tri--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)-(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 dichloromethane (18mL), Deca Dai Si-horse The dichloromethane solution 9.0mmol of the high iodine reagent of fourth, 18h is stirred in reaction at room temperature.With containing 10% sulfur after the completion of question response The aqueous solution of sodium thiosulfate and 5% sodium bicarbonate is quenched reaction, organic faciess saturated common salt water washing, anhydrous sodium sulfate drying, Filter, concentrating under reduced pressure, residue passes through a short silicagel column (CH₂Cl₂:MeOH=10:1 eluting) and concentrate eluant to obtain 2.9g white Color amorphous solid, yield 95%.Through nuclear-magnetism and Mass Spectrometric Identification gained white, amorphous solid be product with its monohydrate and The mixture of dihydrate, by mixture, under high vacuum condition, thermal dehydration can be changed into corresponding product.

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]⁺:Value of calculation C₈₃H₁₀₇N₆O₂₈Si₂, 1691.6629, measured value 1691.6666.

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

Take 1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-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)-(2, 3- bis--O- benzoyl-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 tetrahydrochysene furan In muttering, and it is cooled to -78 DEG C under argon gas atmosphere, be slowly added dropwise the tetrahydrofuran solution of commercially available 1M triisobutyl lithium borohydride 2.2mL (2.2mmol), and react 30min under -78 DEG C of argon gas atmosphere.Reactant liquor glacial acetic acid is quenched, and then subtracts reactant liquor Pressure concentrates, and dissolves residue with ethyl acetate (40mL).Organic faciess use saturated sodium bicarbonate aqueous solution successively, and saturated common salt is washed Wash, aqueous phase is extracted with dichloromethane, merge organic faciess, with anhydrous sodium sulfate drying, filter, be concentrated to give white amorphous powder. This powder is dissolved in a small amount of dichloromethane, Deca ether is to separating out white precipitate under fast stirring, and continues Deca ether extremely Precipitation completely, is collected by centrifugation and precipitates and washed with a small amount of ether, is dried.

This precipitation is placed in 50mL plastic centrifuge tube, adds 10mL oxolane and 10mL pyrido to be cooled to 0 DEG C.Slow Slow Deca HF Pyridine 1mL, after the completion of reactant liquor is slowly increased to room temperature and 24h.TLC display reaction is stirred at room temperature, Reactant liquor is dissolved in Excess ethyl acetate, and uses 10%CuSO successively₄Aqueous solution, 0.01N EDETATE SODIUM saline solution with full And brine It, organic faciess with anhydrous sodium sulfate drying, filter, obtain white, amorphous solid.Gained solid is used no successively 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]⁺:Value of calculation 1450.4983, measured value 1450.5001.

Embodiment 18:1- nitrine -6-O- (beta d glucopyranosiduronic acid base)-(1 → 3)-(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 1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) - (2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(2,3,4- tri--O- benzoyl-β-D- pyrans Portugal Grape alduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane 300mg (1.0equiv.) it is dissolved in dry DMF (4mL) with sulfur trioxide trimethylamine complex (20equiv.).Reactant liquor is at 50 DEG C DEG C After lower reaction 48h, reactant liquor is cooled to room temperature, reactant liquor dropper is added to the glass column of a Sephadex LH-20 filling Upper end (packing volume be (2.5cm × 140cm)), with dichloromethane：Methanol=1：1 eluent, gained flow point warp Silica gel plate ultraviolet 254nm is monitored, and the flow point having fluorescence merges concentration, obtains white foam.This foaming solid is dissolved in THF：H₂O =3：In 1 mixed solution (10mL), be cooled to -10 DEG C, the aqueous solution of 1M LiOH of Deca brand-new and 35% hydrogen peroxide mixed (volume ratio is 2 to close solution：1,5mL), move to room temperature after reactant liquor stirring 1h at -10 DEG C and continue stirring 8h.Reactant liquor is cold But to 0 DEG C.Add methanol (8mL), the aqueous solution (4mL) of 4M NaOH slowly moves to room temperature afterwards, and 12h is stirred at room temperature.Instead Application cationic resin is neutralized to neutrality, filters, with distilled water rinse resin repeatedly, merging filtrate, vacuum distillation is concentrated to dryness, By residue with distilled water eluting, Sephadex LH-20 post desalting and purifying, collect the flow point having product, lyophilizing after merging.? 200mg white foam solid, yield 80%.

Embodiment 19:1- nitrine -6-O- (beta d glucopyranosiduronic acid base)-(1 → 3)-(6- sulfate group -2- deoxidation - 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 1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) - (2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(2,3,4- tri--O- benzoyl-β-D- pyrans Portugal Grape alduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane 300mg (1.0equiv.) it is dissolved in dry DMF (10mL) with sulfur trioxide trimethylamine complex (5equiv.).Reactant liquor is at 40 DEG C After reaction 48h, reactant liquor is cooled to room temperature, reactant liquor dropper is added to the glass column of a Sephadex LH-20 filling Upper end (packing volume is (2.5cm × 140cm)), with dichloromethane：Methanol=1：1 eluent, gained flow point is through silicon Offset plate ultraviolet 254nm is monitored, and the flow point having fluorescence merges concentration, obtains white foam.This foaming solid is dissolved in THF：H₂O= 3：In 1 mixed solution (10mL), it is cooled to -10 DEG C, the aqueous solution of 1M LiOH of Deca brand-new and the mixing of 35% hydrogen peroxide (volume ratio is 2 to solution：1,5mL), move to room temperature after reactant liquor stirring 1h at -10 DEG C and continue stirring 8h.Reactant liquor is cooled down To 0 DEG C.Add 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 cationic resin, filter, with distilled water rinse resin repeatedly, merging filtrate, vacuum distillation is concentrated to dryness, will Residue, with distilled water eluting, Sephadex LH-20 post desalting and purifying, collects the flow point having product, lyophilizing after merging.? 150mg white foam solid, yield 69%.

Embodiment 20:1- nitrine -6-O- (beta d glucopyranosiduronic acid base)-(1 → 3)-(4- sulfate group -2- deoxidation - 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 tetrose

Take 1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) - (2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(2,3,4- tri--O- benzoyl-β-D- pyrans Portugal Grape alduronic acid carbomethoxy)-(1 → 3)-(2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane 190mg (1.0equiv.), 24h is stirred at room temperature in the presence of anhydrous pyridine with Cyanophenacyl (4.0equiv.).Under ice-water bath with Methanol is evaporated to dry, residue absolute ether precipitation after reaction is quenched, precipitation is collected by filtration, with ethyl acetate dissolving, Washed with 1N aqueous hydrochloric acid solution successively, saturation NaHCO₃Water-soluble washing, saturated common salt water washing, anhydrous sodium sulfate drying, filter, Concentrating under reduced pressure obtains weak yellow foam shape solid.This solid and sulfur trioxide trimethylamine complex (10equiv.) are dissolved in anhydrous In DMF (2.5mL).Reactant liquor is cooled to room temperature after reacting 12h at 50 DEG C by reactant liquor, and reactant liquor dropper is added to one The upper end (packing volume is (2.5cm × 140cm)) of the glass column of Sephadex LH-20 filling, with dichloromethane：Methanol= 1：1 eluent, gained flow point is monitored through silica gel plate ultraviolet 254nm, and the flow point having fluorescence merges concentration, obtains white bubble Foam.This foaming solid is dissolved in THF：H₂O=3：In 1 mixed solution (10mL), it is cooled to -10 DEG C, the 1M of Deca brand-new (volume ratio is 2 to the mixed solution of the aqueous solution of LiOH and 35% hydrogen peroxide：1,2mL), reactant liquor moves after stirring 1h at -10 DEG C Continue stirring 8h to room temperature.Reactant liquor is cooled to 0 DEG C.Add methanol (4mL), the aqueous solution (2mL) of 4M NaOH slowly moves afterwards To room temperature, and 12h is stirred at room temperature.Reaction cationic resin is neutralized to neutrality, filters, many with distilled water rinse resin Secondary, merging filtrate, vacuum distillation is concentrated to dryness, by residue with distilled water eluting, Sephadex LH-20 post desalting and purifying, Collect the flow point having product, lyophilizing after merging.Obtain 80mg white foam solid, yield 58%.

Claims

1. a kind of synthetic method of chondroitin sulfate tetrose, methods described comprises the steps：

(1) formula A compound, in the weak acid buffer solution containing sodium chloride, obtains formula Bization in the presence of hyaluronidase Compound；Here, described hyaluronidase derives from animal testis, vigor 400-1000IU/mg；Preferably, selected from bull testis Or sheep testicle；It is highly preferred that being bull testis；

(2) formula B compound is in the alcohol R of mineral acid₁Carry out esterification in OH solution, the carboxyl of formula B compound is esterified； Then utilize anhydride R₂-O-R₂, in the basic conditions remaining all hydroxyl of formula B compound are carried out being acylated protection, obtain formula C Compound；

(3) formula C compound in the presence of a base, 1 of selectively removing reducing end 2-Acetamido-2-deoxy-D-glucose in organic solvent Hydroxyl, obtains formula D compound；

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

Here, alcohol R₄-R₃R in OH₃It is selected from-CH₂CH₂-(OCH₂CH₂) n-, n is 1-100, aliphatic alkane subunit, aliphatic olefin Subunit, or-CH₂-(OCH₂CH₂) m-, m is 1-100；Preferably, R₃For-CH₂-O-CH₂CH₂-；R₄Selected from azido, alkynyl, life Thing element, the amino replacing, aldehyde radical, thioether, described substituted amino refers to the amino protected by protection group, and described protection group is selected from Benzyloxy carbonyl acyl group, tertiary butyloxycarbonyl acyl group, tribromo-acetyl base, trifluoroacetyl group；Preferably, R₄-R₃OH is N₃-CH₂-OCH₂CH₂- OH；

(7) formula G compound is in the alcohol R of acid or alkali₁In OH solution, slough all of R on formula G compound₂, obtain formula H compound；

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

Here, R₅CHO or R₅The R that CHO contracts in two fatty alcohol₅Selected from phenyl or substituted phenyl；

(9) compound of formula I is in carboxylic acid halides R₇X or anhydride R₇-O-R₇In the presence of, it is acylated in the basic conditions, obtained formula J chemical combination Thing；

Here, carboxylic acid halides R₇X or anhydride 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 therefore preferable to chlorine or bromine；

(10) under the existence condition of acid and reducing agent, exposed 4 hydroxyls of selectivity are so that 6 is R to formula J compound₅-CH₂- protect Shield, obtains formula K compound；

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

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

Or, step (10)-(13) are replaced with following step (10a)-(14)：

(10a) formula J compound, under sour existence condition, hydrolyzes two benzals or substituted benzal, obtains formula K ' chemical combination Thing；

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

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

(13a) formula M ' compound is three-dimensional by the ketone carbonyl of 4 and 4 ' positions of 2-Acetamido-2-deoxy-D-glucose under reducing agent existence condition Optionally it is reduced to axial bond hydroxyl, obtain formula N ' compound；

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

(15) utilize formula O compound synthesis chondroitin sulfate tetrose；

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 pi-allyl； Here, described substituted benzyl refers to that phenyl ring is replaced by one or more substituent group, and described substituent group is selected from halogen, nitre Base, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxyl；And, described substituent group is in the optional position of phenyl ring；Preferably Ground, R₁Selected from methyl, ethyl, pi-allyl, benzyl, 4- methoxy-benzyl, it is more preferably selected from methyl, ethyl, benzyl；

In formula C- formula O compound, formula J- formula O compound, R₂And R₇It is each independently selected from aliphatic acyl radical, unsubstituted benzene first Acyl group or substituted benzoyl；Here, described aliphatic acyl radical refers to C2-C6 alkanoyl；Described substituted benzoyl is Finger phenyl ring is replaced by one or more substituent group, described substituent group is selected from halogen, nitro, C1-C4 alkyl, C1-C4 Haloalkyl, C1-C4 alkoxyl；And, described substituent group is in the optional position of phenyl ring；Preferably, R₂And R₇Select independently of one another From acetyl group, benzoyl, 4- chlorobenzoyl base, 4- bromobenzoyl base, it is more preferably selected from acetyl group or benzoyl Base；

In formula G- formula O compound, R₃Selected from-(CH₂CH₂O)n-CH₂CH₂-, n is 1-100 here；Aliphatic alkane subunit；Fat Race's alkene subunit；Or-(CH₂CH₂O)m-CH₂-, m is 1-100 here；Preferably, selected from-CH₂CH₂-O-CH₂-；R₄Selected from nitrine Base, alkynyl, biotin, the amino replacing, aldehyde radical, thioether；Amino that described substituted amino refers to be protected by protection group, described Protection group is selected from benzyloxy carbonyl acyl group, tertiary butyloxycarbonyl acyl group, tribromo-acetyl base, trifluoroacetyl group；Preferably ,-OR₃-R₄For-O- CH₂CH₂-O-CH₂-N₃；

Formulas I, formula J, formula K, in formula L and formula M compound, R₅Selected from unsubstituted phenyl or substituted phenyl, here, described replacement Phenyl refer to that phenyl ring is replaced by one or more substituent group, described substituent group be selected from halogen, nitro, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxyl；And, described substituent group is in the optional position of phenyl ring；Preferably, selected from phenyl, 4- Methoxyphenyl, 4- chlorophenyl or 4- bromo phenyl, it is highly preferred that be selected from phenyl or 4- methoxyphenyl；

Formula L ', formula M ' and formula N ' in compound, R₈And R₉It is each independently selected from C1-C4 alkyl, phenyl or substituted phenyl, Here, described substituted phenyl refers to that phenyl ring is replaced by one or more substituent group, and described substituent group is selected from halogen, nitre Base, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxyl；And, described substituent group is in the optional position of phenyl ring；Preferably Ground, R₈And R₉It is each independently selected from methyl, isopropyl or the tert-butyl group；

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

2. method according to claim 1, wherein, in described step (1), the addition of hyaluronidase is formula A chemical combination The 2.0-3.0% of amount of substance, 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 pH value that described faintly acid refers to buffer solution is 3.0-6.5；Described buffer solution is selected from acetic acid-acetic acid Sodium buffer, acetic acid -50 mM, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer or potassium dihydrogen phosphate-dipotassium hydrogen phosphate One kind of buffer is it is therefore preferable to acetic acid-sodium acetate buffer solution；The concentration of described buffer solution is for 0.01M-1M it is preferable that institute Stating buffer solution for concentration is the acetic acid-sodium acetate buffer solution of 0.1M；Concentration in weak acid buffer solution for the sodium chloride is 0.05M-0.5M is it is preferable that be 0.15M.

3. method according to claim 1, wherein, in described step (2), described mineral acid is selected from hydrogen chloride, sulphuric acid, nitre Acid, phosphoric acid are it is preferable that be selected from hydrogen chloride or sulphuric acid；Described alkalescence condition refers to selected from Sodium Acetate Trihydrate, pyridine, sodium dihydrogen phosphate, The condition that the alkali of potassium carbonate, triethylamine or piperidines exists is it is preferable that refer to the bar existing selected from the alkali of pyridine or Sodium Acetate Trihydrate Part.

4. method according to claim 1, wherein, in described step (3), described alkali is selected from ethylenediamine, hydrazine hydrate or 3- N, N dimethyl amino propylamine is it is preferable that be selected from hydrazine hydrate or 3-N, N dimethyl amino propylamine；Described organic solvent, selected from third Ketone, chloroform, oxolane, dichloromethane, DMF or N, N dimethyl acetamide is it is preferable that be selected from dichloromethane Alkane, oxolane or N, N-dimethylformamide.

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

6. forwarding method according to claim 1, wherein, in described step (5), described acid is selected from trifluoromethanesulfonic acid alkyl silicon Fat, boron trifluoride diethyl etherate, copper chloride, p-methyl benzenesulfonic acid, or camphorsulfonic acid.

7. method according to claim 1, wherein, in described step (6), described acid is selected from trifluoromethanesulfonic acid alkyl silicon Fat, boron trifluoride diethyl etherate, copper chloride, p-methyl benzenesulfonic acid, camphorsulfonic acid；Preferably, selected from copper chloride, trifluoromethanesulfonic acid C1- The silicone grease that C4 trialkyl replaces；Described organic solvent is selected from dichloromethane, chloroform, oxolane, acetone or 1,2- dichloroethanes One of；Preferably, it is chloroform.

8. method according to claim 1, wherein, in described step (7), described acid selected from hydrogen chloride, sulphuric acid, nitric acid, One of phosphoric acid or boron trifluoride diethyl etherate are it is preferable that be selected from hydrogen chloride or boron trifluoride diethyl etherate；Described alkali is selected from R₁ONa、 R₁OK, NaOH are it is therefore preferable to R₁ONa, wherein, R₁Selected from unsubstituted benzyl or substituted benzyl, C1-C4 alkyl or allyl Base；Here, described substituted benzyl refers to that phenyl ring is replaced by one or more substituent group, and described substituent group is selected from halogen Element, nitro, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxyl；And, described substituent group is in the optional position of phenyl ring； Preferably, R₁Selected from methyl, ethyl, pi-allyl, benzyl, 4- methoxy-benzyl, it is more preferably selected from methyl, ethyl, benzyl；

Preferably, in described step (8), described acid is selected from p-methyl benzenesulfonic acid or camphorsulfonic acid；

Preferably, in described step (9), described alkalescence condition refers to selected from Sodium Acetate Trihydrate, pyridine, sodium dihydrogen phosphate, potassium carbonate, The condition that the alkali of triethylamine or piperidines exists is it is preferable that refer to the condition existing selected from the alkali of pyridine or Sodium Acetate Trihydrate；

Preferably, in described step (10), described acid is selected from one of trifluoroacetic acid, acetic acid, boric acid it is preferable that being trifluoro Acetic acid；Described reducing agent be selected from trialkyl replace silane, sodium cyanoborohydride, borine it is preferable that selected from trialkyl replace Silane or sodium cyanoborohydride；

Preferably, in described step (11), described oxidant is selected from Dai Si-Martin's oxidant, the group of anhydride-dimethyl sulfoxide Conjunction, the combination of the chloro- dimethyl sulfoxide-triethylamine of oxalyl, the combination of trifluoroacetic anhydride-dimethyl sulfoxide-triethylamine or titanium dioxide Manganese；Preferably, it is Dai Si-Martin's oxidant；

Preferably, in described step (12), described reducing agent is selected from sodium borohydride, lithium borohydride, potassium borohydride or alkyl and replaces Borohydride salts；Preferably, selected from triisobutyl potassium borohydride or sodium borohydride；

Preferably, in described step (13), described oxidant is selected from DDQ or ammonium ceric nitrate；Described reducing agent is selected from triethyl group silicon Combining of alkane and iodine or combining of triethyl silicane and carbon monoxide and cobalt octacarbonyl；

Preferably, in described step (10a), described acid is selected from one of trifluoroacetic acid, acetic acid, boric acid it is preferable that being second Acid；

Preferably, in described step (11a), described substituted chlorosilane is selected from tert-butyl diphenyl chlorosilane, tert-butyl group diformazan Base chlorosilane, tri isopropyl chlorosilane, chlorotriethyl silane, 3,5-dimethylphenyl chlorosilane or trim,ethylchlorosilane；Preferably, it is Tert-butyl chloro-silicane；Described alkali is selected from triethylamine, pyridine or DMAP；Preferably, it is pyridine；

Preferably, in described step (12a), described oxidant is selected from the combination of the chloro- dimethyl sulfoxide-triethylamine of oxalyl, acetic acid The combination of acid anhydride-dimethyl sulfoxide, manganese dioxide or the high iodine reagent of Dai Si-Martin；It is preferably the high iodine reagent of Dai Si-Martin；

Preferably, in described step (13a), described reducing agent is selected from sodium borohydride, lithium borohydride, potassium borohydride or alkyl and replaces Borohydride salts；Preferably, selected from triisobutyl potassium borohydride or sodium borohydride；

Preferably, in described step (14), described acid is selected from p-methyl benzenesulfonic acid or trifluoroacetic acid it is therefore preferable to p-methyl benzenesulfonic acid； Described fluorion reagent is selected from tetrabutyl ammonium fluoride, hydrogen fluoride pyridine complex, fluohydric acid gas triethylamine complex or ammonium fluoride；Excellent Selection of land is hydrogen fluoride pyridine complex.

9. it is used for synthesizing the midbody compound of chondroitin sulfate tetrose, including following formula D- formula N ' compound：

Wherein, R₁,R₂,R₃,R₄,R₅,R₇,R₈,R₉Definition as described in the appended claim 1.

10. midbody compound as claimed in claim 9, including：

(2,3,4- tri--O- acetyl group-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6- bis--O- acetyl group -2- takes off Oxygen-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-beta d glucopyranosiduronic acid first Ester group)-(1 → 3) -4,6- two-O- acetyl group -2- '-deoxy-n-acetylaminohydroxyphenylarsonic acid D- Glucopyranose.；

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

(4,6- bis--O- acetyl group -2- takes off O- (2,3,4- tri--O- acetyl group-β-D-Glucose aldehydic acid carbomethoxy)-(1 → 3) - Oxygen-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D-Glucose aldehydic acid methyl ester Base)-(1 → 3) -2- methyl-(4,6- bis--O- acetyl group -1,2- dideoxy-α-D- Glucopyranose .) [2,1-d] 2- oxazoline；

1- nitrine -2- oxa- -4-O- [(2,3,4- tri--O- acetyl group-β-D-Glucose aldehydic acid methyl ester)-(1 → 3)-(4,6- bis- - O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D- Portugal Grape alduronic acid carbomethoxy)-(1 → 3)-(4,6- bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- Glucopyranose. 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；

[(beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- is to ar-methoxy-benzylidene -2- for 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；

1- nitrine -2- oxa- -4-O- [(2,3,4- tri--O- benzoyl-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- bis--O- benzene 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；

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

1- nitrine -2- oxa- -4-O- [(2,3,4- tri--O- benzoyl-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- bis--O- 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；

1- nitrine -2- oxa- -4-O- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3) - (6-O- is to methoxybenzyl -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)-(1 → 4)-(2,3- bis--O- benzoyl 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- [(2,3,4- tri--O- acetyl group-β-D-Glucose aldehydic acid methyl ester)-(1 → 3)-(4,6- bis--O- acetyl Base -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- acetyl group-β-D-Glucose aldehyde Sour carbomethoxy)-(1 → 3)-(4,6- bis--O- acetyl group -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid α-D- glucopyranosyl)] 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- [(2,3,4- tri--O- benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(4,6-O- Benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- glucopyranosyl)-(1 → 4)-(2,3- bis--O- benzoyl-β-D- pyrrole Glucopyranoside aldehydic acid carbomethoxy)-(1 → 3)-(4,6-O- benzal -2- deoxidation -2-N- acetylaminohydroxyphenylarsonic acid β-D- Glucopyranose. Base)] hexane；

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

1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-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)-(2,3- bis--O- benzene first 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- [(2,3,4- tri--O- benzoyl-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)-(2,3- bis--O- Benzoyl-beta d glucopyranosiduronic acid carbomethoxy)-(1 → 3)-(6-O- dimethyl tertiary butyl silicon ether -4- ketone -2- takes off Oxygen -2-N- acetylaminohydroxyphenylarsonic acid β-D- galactopyranosyl glycosyl)] hexane；

1- nitrine -6-O- [(2,3,4- tri--O- benzoyl-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)-(2,3- bis--O- benzene first 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

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