CN106432371A - Beta-1,2-D-oligomeric mannoprotein conjugates and preparation method and application thereof - Google Patents

Beta-1,2-D-oligomeric mannoprotein conjugates and preparation method and application thereof Download PDF

Info

Publication number
CN106432371A
CN106432371A CN201610928843.7A CN201610928843A CN106432371A CN 106432371 A CN106432371 A CN 106432371A CN 201610928843 A CN201610928843 A CN 201610928843A CN 106432371 A CN106432371 A CN 106432371A
Authority
CN
China
Prior art keywords
och
oligomerization
conjugate
mannoproteins
mannose
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201610928843.7A
Other languages
Chinese (zh)
Inventor
廖俊
何大为
刘超
吴也
付奔
潘炜华
吴秋业
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Second Military Medical University SMMU
Original Assignee
Second Military Medical University SMMU
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Second Military Medical University SMMU filed Critical Second Military Medical University SMMU
Priority to CN201610928843.7A priority Critical patent/CN106432371A/en
Publication of CN106432371A publication Critical patent/CN106432371A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
    • C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

The invention discloses beta-1,2-D-oligomeric mannoprotein conjugates shown in the formula I in the description and a preparation method and application thereof. In the formula, n is 0 or 1 or 2, and R is carrier protein KLH or HSA. According to the preparation method, phosphorylated beta-1,2-D-oligomeric mannan oligosaccharides are synthesized with alpha-D-glucose as the raw material and then are coupled with the carrier protein, and the mannoprotein conjugates are obtained. When the prepared compounds are used for immunizing mice as an anti-candida albicans vaccine, the result shows that the compounds can induce the body to make a strong immune response; the beta-1,2-D-manninotriose protein conjugate has the highest immunogenicity, generated immune serum can be used for specifically recognizing candida albicans cell surface antigens, and thus the purpose of preventing candida albicans infection can be achieved. The beta-1,2-D-oligomeric mannoprotein conjugates have high application value.

Description

β -1,2-D- oligomerization Mannoproteins conjugate and its preparation method and application
Technical field
The present invention relates to antimycotic saccharide vaccines technical field is and in particular to β -1,2-D- oligomerization Mannoproteins conjugate and Its preparation method, the application in the prevention and treatment of infection by Candida albicans as vaccine of such glycoprotein conjugate.
Background technology
In recent years, the death rate that infection by Candida albicans causes constantly rises, and germ is serious for antibacterials generation Drug resistance, clinically needs effectively to prevent and treat medicine and the strategy of infection by Candida albicans.The immunization therapy of fungal infection Mainly take is Activeimmunotberapy, that is, utilize antigenic stimulus body to produce powerful immune response, by producing antibody Or differentiation effector cell reaches the fungi removing infection or the purpose of other microorganism, there is toxic and side effect little, evident in efficacy etc. Feature.β -1,2- mannosan is the T- cell dependent type polysaccharide antigen that there is candida albicans cell surface, generally by they with Carrier protein (BSA/TT) is coupled and forms glucoprotein vaccine, and research finds that this kind of vaccine can induce body to produce protection antibody, But the immune response causing is limited.β -1,2- mannobiose, trisaccharide and tetrose that we plan to synthesize pass through phosphoethanolamine base Group's (natural glycoprotein connected mode) and strongly immunogenic carrier protein keyhole worm Qi Xue azurin (KLH) are coupled and form sugared egg White conjugate, is desirably to obtain the glucoprotein vaccine of high immunogenicity.
Content of the invention
First purpose of the present invention is for deficiency of the prior art, provides a kind of β -1,2-D- oligomerization mannose egg White conjugate.
Second object of the present invention is to provide β -1 as described above, the preparation side of 2-D- oligomerization Mannoproteins conjugate Method.
Third object of the present invention is to provide β -1 as described above, the purposes of 2-D- oligomerization Mannoproteins conjugate.
Fourth object of the present invention is to provide a kind of antifungal fungi vaccine.
For realizing above-mentioned first purpose, the present invention adopts the technical scheme that：
A kind of β -1,2-D- oligomerization Mannoproteins conjugate, described β -1,2-D- oligomerization Mannoproteins conjugate structure Formula is as follows：
Wherein, n is carrier protein KLH or HSA for 0,1 or 2, R.
Further, the monosaccharide unit of described composition oligosaccharides is β -1,2-D- mannose.
Further, described β -1,2-D- oligomerization Mannoproteins conjugate by β -1 of phosphorylation, 2-D- oligomerization mannose with Carrier protein couplet is obtained.
Further, described β -1,2-D- oligomerization Mannoproteins conjugate is β -1 using glutaryl as phosphorylation, 2- D- oligomerization mannose and the linking group of carrier protein.
For realizing above-mentioned second purpose, the present invention adopts the technical scheme that：
β -1, the preparation method of 2-D- oligomerization Mannoproteins conjugate, comprise the steps as mentioned above：
The first step, synthesizes β -1,2-D- mannobiose acceptor 7, β -1,2-D- manninotriose acceptor 10：
Reaction condition：A) TMSOTf, CH2Cl2,Molecular sieve, -40 DEG C, 92.4%；b)CH3ONa, CH3OH, 94.2%； C) i) DMSO, Ac2O；Ii) L-selectride, THF, -78 DEG C, 79.5% (two step total recoverys)；D) TMSOTf, CH2Cl2, Molecular sieve, -40 DEG C, 90.3%；e)CH3ONa, CH3OH, 92.7%；F) i) DMSO, Ac2O；Ii) L-selectride, THF ,- 78 DEG C, 77.6% (two step total recoverys).
Second step, synthesizes phosphorylation β -1,2-D- oligomerization mannose 19a-c：
Reaction condition：A) TMSOTf, CH2Cl2,Molecular sieve, -40 DEG C, (12a, 84.3%；12b, 89.5%；12c, 81.5%)；b)CH3ONa, CH3OH, 91.3-95.5%；C) i) DMSO, Ac2O；Ii) L-selectride, THF, -78 DEG C, 79.6-84.5% (two step total recoverys)；D) BnBr, NaH, TBAI, 0 DEG C, 83.7-89.4%；E) TBAF, THF, 83.4- 88.2%；f)i)Tetrazole,CH2Cl2/CH3CN；Ii) tert-BuOOH, 79.3-87.5% (two step total recoverys)；G) DBU, CH2Cl2, 86.3-93.2%.
3rd step, synthesizes β -1,2-D- oligomerization Mannoproteins conjugate 1a-c, 2a-c：
Reaction condition:a)H2, Pd (OH)2/ C, CH2Cl2/MeOH(1：1, v/v), 24 hours, 88.7-93.5%；b)DMF/ PBS(4：1, v/v), 4 hours；C) PBS, 2.5 days.
Further, described phosphorylation β -1,2-D- oligomerization mannose preparation method is as follows：
(1) with alpha-D-glucose as raw material, preparation β -1 in the presence of reaction promoter, 2-D- sweet dew monose acceptor and 3, 4,6- tribenzyl -2- acetoxyl group glucose tri- chloroacetimidate donor；
(2) there is glycosylation system in the monose donor of above-mentioned preparation and monose acceptor under catalyst (TMSOTf) effect Standby disaccharides, prepares β -1,2-D- mannobiose acceptor by configuration reversal on 2 '-position of disaccharides；β -1,2-D- mannobiose is subject to Body and monose donor prepare β -1,2-D- manninotriose acceptor by preceding method；
(3) β -1,2-D- mannose receptor respectively with 6- tert-butyl diphenyl silicon substrate -3,4- dibenzyl -2- acetoxyl group Portugal Grape sugar tri- chloroacetimidate donor prepares corresponding β -1,2-D- oligomerization mannose through glycosylation；Oligomerization mannose is non-reduced React and phosphorylation β -1,2-D- oligomerization mannose is obtained with phosphorylation agent after TBDPS protection is taken off in end 6- position.
Further, described phosphorylation β -1,2-D- oligomerization mannose preparation method comprises the steps：
(1) β -1,2-D- oligomerization mannose of phosphorylation and hydroxyl Succinimidyl glutarate are reacted and β -1,2-D- is obtained Oligomerization mannose Acibenzolar；
(2) β -1,2-D- oligomerization mannose Acibenzolar preparing step (1) is even with carrier protein under weak basic condition Connection, obtains β -1,2-D- oligomerization Mannoproteins conjugate.
For realizing above-mentioned 3rd purpose, the present invention adopts the technical scheme that：
As above arbitrary described β -1,2-D- oligomerization Mannoproteins conjugate is in preparation prevention and/or treatment fungal infection Medicine in application.Described fungi is candida albicans.
For realizing above-mentioned 4th purpose, the present invention adopts the technical scheme that：
A kind of antifungal fungi vaccine, including β -1 of therapeutically effective amount as described above, 2-D- oligomerization Mannoproteins Conjugate and pharmaceutically acceptable auxiliary material or adjuvant composition.
The invention has the advantages that：
1st, using the compound of preparation as antifungal vaccine immune mouse, result shows that the compound of the present invention can lure Lead body and produce stronger immune response, wherein β -1, the immunogenicity of 2-D- manninotriose protein conjugate is the strongest and generation Immune serum can specific recognition candida albicans cell surface antigen, such that it is able to play prevention infection by Candida albicans work With, β -1 of the present invention, 2-D- oligomerization Mannoproteins conjugate has stronger using value.
Brief description
Accompanying drawing 1 is β -1,2-D- oligomerization Mannoproteins conjugate formula.
Accompanying drawing 2 is β -1,2-D- mannobiose, β -1,2-D- manninotriose acceptor synthetic route.Reaction condition：a) TMSOTf, CH2Cl2,Molecular sieve, -40 DEG C, 92.4%；b)CH3ONa, CH3OH, 94.2%；C) i) DMSO, Ac2O；ii)L- Selectride, THF, -78 DEG C, 79.5% (two step total recoverys)；D) TMSOTf, CH2Cl2,Molecular sieve, -40 DEG C, 90.3%；e)CH3ONa, CH3OH, 92.7%；F) i) DMSO, Ac2O；Ii) L-selectride, THF, -78 DEG C, 77.6% (two Step total recovery).
Accompanying drawing 3 is phosphorylation β -1,2-D- oligomerization mannose disaccharides, trisaccharide synthetic route.Reaction condition：A) TMSOTf, CH2Cl2,Molecular sieve, -40 DEG C, (12a, 84.3%；12b, 89.5%；12c, 81.5%)；b)CH3ONa, CH3OH, 91.3- 95.5%；C) i) DMSO, Ac2O；Ii) L-selectride, THF, -78 DEG C, 79.6-84.5% (two step total recoverys)；d) BnBr, NaH, TBAI, 0 DEG C, 83.7-89.4%；E) TBAF, THF, 83.4-88.2%；f)i)Tetrazole,CH2Cl2/ CH3CN；Ii) tert-BuOOH, 79.3-87.5% (two step total recoverys)；G) DBU, CH2Cl2, 86.3-93.2%.
Accompanying drawing 4 is β -1,2-D- oligomerization mannose disaccharides, trisaccharide, tetrose protein conjugate synthetic route.Reaction condition:a) H2, Pd (OH)2/ C, CH2Cl2/MeOH(1：1, v/v), 24 hours, 88.7-93.5%；b)DMF/PBS(4：1, v/v), 4 hours； C) PBS, 2.5 days.
Accompanying drawing 5 is the MALDI-TOF-MS spectrogram of compound 2a.
Accompanying drawing 6 is the MALDI-TOF-MS spectrogram of compound 2b.
Accompanying drawing 7 is the MALDI-TOF-MS spectrogram of compound 2c.
Accompanying drawing 8 is total antibody titer in detection serum after compound 1a-c immune mouse, and what wherein+Adj represented is to add Adjuvant group.
Accompanying drawing 9 is the immunofluorescence that gained serum is combined with candida albicans cell after compound 1b third time immune mouse Figure.
Accompanying drawing 10 is the fluidic cell that gained serum is combined with candida albicans cell after compound 1b third time immune mouse Figure.
Specific embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate this Bright rather than limit the scope of the present invention.In addition, it is to be understood that after having read the content of present invention record, art technology Personnel can make various changes or modifications to the present invention, and these equivalent form of values equally fall within the application appended claims and limited Fixed scope.
β -1,2-D- oligomerization Mannoproteins conjugate (general structure is as shown in Figure 1) of the present invention prepare reaction scheme (Fig. 2, Fig. 3, Fig. 4)：
The first step, synthesizes β -1,2-D- mannobiose (acceptor 7), β -1,2-D- manninotriose acceptor (acceptor 10)：
Second step, synthesizes phosphorylation β -1,2-D- oligomerization mannose 19a-c：
3rd step, synthesizes glycoprotein conjugate 1a-c, 2a-c：
The synthesis of embodiment 1 compound 5
By monose acceptor 3 (1.23g, 2.5mmol) (J.Org.Chem.2001,66,8411.), donor 4 (1.95g, 3.0mmol) (J.Carbohydr.Chem.1994,13,421.) and activationMolecular sieve (1.0g) is placed in 100mL round bottom Flask is simultaneously vacuum dried half an hour, then dissolves mixture 50mL anhydrous methylene chloride.Suspension argon gas protection under in Under room temperature stirring half an hour after be cooled to -40 DEG C, be then slowly added dropwise trimethylsilyl trifluoromethanesulfonate (35.0 μ L, 0.18mmol).After half an hour, the neutralization of reactant liquor triethylamine is quenched and is filtered with diatomite, and after concentration, gained head product is through silica gel Column chromatography (n-hexane/ethyl acetate, 12:1, v/v) obtain white foam solid (2.23g, 92.4%).
1HNMR(400MHz,CDCl3)δ7.40-7.19(m,30H,Ar),5.88(m,1H,OCH2CH=CH2),5.37(m, 1H,OCH2CH=CH 2),5.24-5.20(m,1H,OCH2CH=CH 2), 5.15 (dd, J=7.6Hz, 9.6Hz, 1H, H-2 '), 4.86-4.75(m,6H,H-1’,CH2Ph),4.56-4.43(m,7H,CH2Ph),4.42-4.39(m,1H,OCH 2CH=CH2), 4.34 (s, 1H, H-1), 4.28 (d, J=3.2Hz, 1H, H-2), 4.22-4.01 (m, 1H, OCH 2CH=CH2),3.82-3.74 (m, 3H, H-3, H-6, H-6 '), 3.73-3.57 (m, 5H, H-4, H-4 ', H-5 ', H-6 '), 3.52 (d, J=3.2Hz, H-3), 3.44(m,1H),1.97(s,3H,Ac).ESI-MS:calcd.for C59H64O12[M+Na]+m/z,987.4；found, 987.9.
The synthesis of embodiment 2 compound 6
Compound 5 (1.85g, 2.0mmol) is used 100mL absolute methanol to dissolve, addition sodium methoxide (11.0mg, 0.2mmol) juxtaposition is stirred overnight at room temperature, adds ion exchange resin IR 120 (H+form) neutralization, mistake after completion of the reaction Filter resin, after concentration gained head product through silica gel column chromatography (n-hexane/ethyl acetate, 4:1, v/v) obtain white foam Solid (1.74g, 94.2%).
1H NMR(400MHz,CDCl3)δ7.55-7.23(m,30H,Ar),6.05-5.93(m,1H,OCH2CH=CH2), 5.40(m,1H,OCH2CH=CH 2),5.29(m,1H,OCH2CH=CH 2), 5.17 (dd, J=11.2Hz, 1H, OCH2Ph), 5.05-4.93(m,4H,OCH2), Ph 4.90 (d, J=11.2Hz, 1H, OCH2Ph), 4.81 (d, J=7.6Hz, 1H, H-1 '), 4.72 (d, J=12.0Hz, 1H, OCH2Ph),4.67-4.46(m,10H,OCH2Ph,OCH 2CH=CH2, H-1), 4.36 (d, J= 3.2Hz,1H,H-2),4.17-4.10(m,1H,OCH 2CH=CH2), 4.01 (t, J=9.6Hz, 1H, H-4), 3.88-3.71 (m, 6H,H-2’,H-3’,H-6a’,H-6b’,H-6a,H-6b),3.70-3.57(m,3H,H-3,H-4’,H-5’),3.53-3.42 (m,1H,H-5).13C NMR(100MHz,CDCl3)δ139.13,138.52,138.33,138.27,138.16,138.11, 133.52,128.78,128.56,128.45,128.41,128.36,128.23,128.17,128.12,128.03,127.98, 127.93,127.87,127.80,127.72,127.70,127.66,127.54,117.86,104.11,99.39,85.17, 80.33,77.30,75.70,75.46,75.34,75.11,74.78,74.63,74.37,73.48,70.39,69.97, 69.81,69.23.ESI-MS:calcd.for C57H62O11[M+K]+m/z,962.2；found,930.5.
The synthesis of embodiment 3 compound 7
Compound 6 (1.38g, 1.5mmol) is placed in 100mL round-bottomed flask, is subsequently adding dimethyl sulfoxide (DMSO) (22.0mL) With aceticanhydride (11.0mL).Reactant liquor is extracted with ethyl acetate after being stirred at room temperature 18 hours, respectively use saturated sodium carbonate and Brine It, then uses anhydrous sodium sulfate drying, and the head product being concentrated to give after filtration toluene band revolves twice.Residue is used 40mL anhydrous tetrahydro furan dissolves and is cooled to -78 DEG C, is slowly added dropwise L-Selectride (1 under argon gas protectionMTHF,7.5mL) And stir 15 minutes, then remove cooling device juxtaposition and continue at room temperature to stir 15 minutes, methyl alcohol adds after reaction is quenched Enter 50mL dchloromethane, solution uses hydrogenperoxide steam generator (5%, 30mL), sodium hydroxide solution (1 respectivelyM, 30mL), sulphur Sodium thiosulfate solution (5%, 30mL) and saturated aqueous common salt (30mL) washing, after being dried through anhydrous magnesium sulfate, filter and concentrating Pale yellow oily liquid, gained head product through silica gel column chromatography (n-hexane/ethyl acetate, 4:1, v/v) obtain white foam solid Body (1.10g, 79.5%).
1H NMR(400MHz,CDCl3)δ7.55-7.23(m,30H,Ar),5.99-5.86(m,1H,OCH2CH=CH2), 5.27(m,1H,OCH2CH=CH 2),5.21(m,1H,OCH2CH=CH 2), 5.03 (s, 1H, H-1 '), 5.02 (dd, J= 10.8Hz,1H,OCH2), Ph 4.99 (dd, J=11.2Hz, 1H, OCH2), Ph 4.94 (dd, J=10.8Hz, 1H, OCH2Ph), 4.89 (dd, J=10.8Hz, 1H, OCH2), Ph 4.69 (dd, J=10.8Hz, 1H, OCH2Ph), 4.68 (dd, J=11.6Hz, 1H,OCH2Ph),4.64-4.59(m,2H,OCH2Ph), 4.57 (d, J=3.2Hz, 1H, H-2), 4.56-4.48 (m, 4H, OCH2Ph),4.47(s,1H,H-1),4.46-4.43(m,1H,OCH 2CH=CH2), 4.41 (d, J=3.2Hz, 1H, H-2 '), 4.11-4.06(m,1H,OCH 2CH=CH2), 3.98 (t, J=9.2Hz, 1H, H-4 '), 3.91-3.70 (m, 5H, H-6a, H-4, H-6a’,H-6b,H-6b’),3.67-3.54(m,3H,H-3’,H-3,H-5’),3.51-3.44(m,1H,H-5).13C NMR (100MHz,CDCl3)δ138.44,138.31,138.18,133.83,128.38,128.27,128.16,128.01, 127.95,127.78,127.72,127.65,117.18,100.15,99.35,81.42,80.34,75.6,75.14,74.44, 74.07,73.50,73.39,70.85,70.75,70.07,69.40,67.70.ESI-MS:calcd.for C57H62O11[M+ Li]+m/z,930.1；found,930.5.
The synthesis of embodiment 4 compound 8
By two saccharide acceptors 7 (0.95g, 1.0mmol), monose donor 4 (0.78g, 1.2mmol) and activationMolecule Sieve (0.25g) is placed in 50mL round-bottomed flask and is vacuum dried half an hour, then dissolves mixture 30mL anhydrous methylene chloride. Suspension is cooled to -40 DEG C after stirring half an hour at room temperature under argon gas protection, is then slowly added dropwise trimethyl silicon substrate trifluoro Methanesulfonates (14.0 μ L, 0.075mmol).After half an hour, the neutralization of reactant liquor triethylamine is quenched and is filtered with diatomite, concentrates Afterwards gained head product through silica gel column chromatography (n-hexane/ethyl acetate, 12:1, v/v) white foam solid (1.26g, 90.3%).
1H NMR(600MHz,CDCl3)δ7.42-7.02(m,45H,Ar),5.86(m,1H,OCH2CH=CH2),5.26 (d, J=8.0Hz, 1H, H-1 "), and 5.20-5.17 (m, 2H, H-2 ", OCH2CH=CH 2),5.11-5.07(m,1H,OCH2CH= CH 2),4.98-4.94(m,3H,OCH2Ph),4.85-4.80(m,2H,OCH2Ph),4.75-4.69(m,4H,H-1’, OCH2), Ph 4.63 (d, J=13.2Hz, 1H, OCH2Ph),4.56-4.47(m,9H,H-2’,OCH2), Ph 4.42 (d, J= 12.0Hz,1H,OCH2Ph),4.41(s,1H,H-1),4.38-4.35(m,1H,OCH2CH=CH2), 4.20 (d, J=3.0Hz, 1H, H-2), 3.91 (t, J=8.4Hz, 1H, H-3 "), and 3.82-3.66 (m, 8H, H-4 ", H-5 ', H-5 ", H-6a " and, H-6a, H- 6b, H-6a ', H-6b '), 3.62 (t, J=9.6Hz, 1H, H-4 '), 3.54-3.46 (m, 3H, H-3, H-3 ', H-6a "), 3.37 (m,1H,H-5).MALDI-TOF-MS:calcd.for C86H92O17[M+Na]+m/z,1419.6,found 1420.0.
The synthesis of embodiment 5 compound 9
Compound 8 (1.02g, 0.75mmol) is used 50mL absolute methanol to dissolve, addition sodium methoxide (11.0mg, 0.2mmol) juxtaposition is stirred overnight at room temperature, adds ion exchange resin IR 120 (H+form) neutralization, mistake after completion of the reaction Filter resin, after concentration gained head product through silica gel column chromatography (n-hexane/ethyl acetate, 6:1, v/v) obtain white foam Solid (0.94g, 92.7%).
1H NMR(400MHz,CDCl3)δ7.56-7.08(m,45H,Ar),6.01-5.89(m,1H,OCH2CH=CH2), 5.29(m,1H,OCH2CH=CH 2),5.24(m,1H,OCH2CH=CH 2),5.16-5.02(m,6H,OCH2Ph,H-1’),4.96 (d, J=10.8Hz, 1H, OCH2Ph), 4.88 (d, J=8.0Hz, 1H, H-1 "), 4.81-4.66 (m, 3H, OCH2Ph),4.65- 4.41(m,13H,OCH2Ph,H-2’,H-2,OCH 2CH=CH2,H-1),4.15-4.07(m,1H,OCH 2CH=CH2),4.01- 3.71(m,9H,H-2”,H-3”,H-4’,H-6b”,H-6a”,H-6a,H-6b,H-6a’,H-6b’),3.68-3.56(m,5H, 1H,H-5”,H-3’,H-4”,H-3,H-5),3.52-3.44(m,1H,H-5’).13C NMR(100MHz,CDCl3)δ139.21, 138.68,138.59,138.44,138.39,138.16,138.11,133.79,128.50,128.41,128.36,128.31, 128.25,128.20,128.11,128.06,127.92,127.86,127.83,127.73,127.69,127.64,127.59, 127.48,127.43,127.27,117.43,105.35,100.08,99.93,86.73,80.25,80.03,77.18, 75.52,75.44,75.31,75.25,74.99,74.83,74.72,74.62,74.09,73.66,73.46,73.33, 71.16,70.31,70.19,69.77,69.45.MALDI-TOF-MS:calcd.for C84H90O16[M+Na]+m/z, 1378.617；found,1378.115.
The synthesis of embodiment 6 compound 10
Compound 9 (678.0mg, 0.5mmol) is placed in 50mL round-bottomed flask, is subsequently adding dimethyl sulfoxide (DMSO) (7.0mL) With aceticanhydride (3.5mL).Reactant liquor is extracted with ethyl acetate after being stirred at room temperature 18 hours, uses saturated sodium carbonate and food respectively Salt water washing, then uses anhydrous sodium sulfate drying, and the head product being concentrated to give after filtration toluene band revolves twice.Residue is used 20mL anhydrous tetrahydro furan dissolves and is cooled to -78 DEG C, is slowly added dropwise L-Selectride (1 under argon gas protectionMTHF, 2.45mL) and stir 15 minutes, then remove cooling device juxtaposition and continue at room temperature to stir 15 minutes, methyl alcohol is quenched instead Should after add 20mL dchloromethane, solution respectively use hydrogenperoxide steam generator (5%, 20mL), sodium hydroxide solution (1M, 20mL), hypo solution (5%, 20mL) and saturated aqueous common salt (20mL) washing, through anhydrous magnesium sulfate be dried, filter and Pale yellow oily liquid after concentration, gained head product through silica gel column chromatography (n-hexane/ethyl acetate, 6:1, v/v) must be white Foaming solid (526.3mg, 77.6%).
1H NMR(400MHz,CDCl3)δ7.66-6.94(m,45H,Ar),5.95-5.84(m,1H,OCH2CH=CH2), 5.25(m,1H,OCH2CH=CH 2),5.20(s,1H,H-1”),5.19(m,1H,OCH2CH=CH 2),5.17(s,1H,H-1’), 5.06-4.96(m,4H,OCH2), Ph 4.90 (d, J=10.8Hz, 1H, OCH2Ph),4.75(br s,1H,H-2’),4.71- 4.64(m,2H,OCH2Ph,H-2),4.61-4.42(m,10H,OCH2Ph,OCH 2CH=CH2, H-1), 4.38 (d, J= 10.8Hz,1H,OCH2Ph),4.35(br s,1H,H-2”),4.28-4.21(m,1H,OCH2Ph),4.10-3.89(m,4H,H- 4”,H-4’,OCH2Ph),3.88-3.49(m,13H,H-6a”,H-6b”,H-5”,H-3”,H-6a’,H-6b’,H-5’,H-3’, H-6a,H-6b,H-5,H-4,H-3),3.47-3.41(m,1H,H-5).13C NMR(100MHz,CDCl3)δ138.56, 138.48,138.26,138.20,138.15,138.10,138.02,137.88,133.62,129.16,128.44,128.38, 128.35,128.29,128.25,128.19,128.11,128.05,127.94,127.79,127.74,127.69,127.65, 127.60,127.51,127.45,127.34,127.18,117.42,100.69,100.06,83.12,80.48,80.35, 75.42,75.35,75.22,75.17,75.07,74.78,74.43,74.34,73.52,73.44,73.36,71.38, 70.34,70.09,70.03,69.95,69.80,69.67,69.49,69.42,68.93,67.29.MALDI-TOF-MS: calcd.for C84H90O16[M+Na]+m/z,1378.617；found,1379.306.
The synthesis of embodiment 7 compound 12a
By monose acceptor 3 (245mg, 0.5mmol), donor 11 (471.0mg, 0.6mmol) (Eur.J.Org.Chem.1999,2523.) and activationMolecular sieve (200mg) is placed in 25mL round-bottomed flask and vacuum is done Dry half an hour, then mixture 10mL anhydrous methylene chloride is dissolved.Suspension stirs half under argon gas protection at room temperature It is cooled to -40 DEG C after hour, be then slowly added dropwise trimethylsilyl trifluoromethanesulfonate (6.35 μ L, 0.035mmol).Half an hour The neutralization of reactant liquor triethylamine is quenched and is filtered with diatomite afterwards, and after concentration, gained head product is through silica gel column chromatography (n-hexane/second Acetoacetic ester, 18:1, v/v) obtain white foam solid (478.5mg, 84.3%).
1H NMR(600MHz,CDCl3)δ7.71-7.01(m,35H,Ar),5.99-5.91(m,1H,OCH2CH=CH2), 5.36(m,1H,OCH2CH=CH 2),5.21(m,1H,OCH2CH=CH 2), 5.16 (dd, J=9.0,8.0Hz, 1H, H-2 '), 5.02 (d, J=8.0Hz, 1H, H-1 '), 4.97-4.91 (m, 2H, OCH2Ph),4.81-4.73(m,3H,OCH2Ph),4.59(br s,2H),4.49-4.41(m,5H,OCH2Ph,OCH 2CH=CH2, H-1), 4.38 (d, J=3.6Hz, 1H, H-2), 4.08-4.01 (m,2H,OCH 2CH=CH2, H-6a '), 3.65-3.60 (m, 1H, H-6b '), 3.79 (br d, J=9.6Hz, 1H, H-6a), 3.75 (t, J=9.0Hz, 1H, H-3 '), and 3.64-3.59 (m, 2H, H-6b, H-4), 3.56-3.49 (m, 4H, H-3, H-4 ', H- 5’),3.48-3.43(m,1H,H-5),1.95(s,3H),1.04(s,9H).13C NMR(150MHz,CDCl3)δ169.9, 138.5,138.4,137.9,137.7,135.7,135.6,133.9,133.4,133.3,129.7,129.6,128.3, 128.28,128.25,128.2,128.0,127.9,127.8,127.7,127.69,127.65,127.6,127.5,127.4, 116.5,100.9,99.8,83.2,79.8,77.9,76.6,75.5,75.2,74.8,74.7,74.6,73.5,73.2,71.7, 70.4,69.41,69.37,64.0,26.9,21.4,19.3.HRMS(ESI):calcd.for C68H76O12Si[M+Na]+m/z, 1135.5004；found,1135.4979.
The synthesis of embodiment 8 compound 12b
Prepared by compound 7 and compound 11, concrete operation step is with the synthesis of compound 12a, yield 89.5%.
1H NMR(600MHz,CDCl3)δ7.64-6.87(m,50H,Ar),5.87-5.78(m,1H,OCH2CH=CH2), 5.36 (d, J=8.4Hz, 1H, H-1 "), 5.21 (dd, J=9.0,8.4Hz, 1H, H-2 "), 5.20 (br d, J=10.8Hz, 1H,OCH2CH=CH 2), 5.09 (br d, J=10.2Hz, 1H, OCH2CH=CH 2),4.98-4.89(m,3H,OCH2Ph),4.85 (d, J=12.0Hz, 1H, OCH2Ph),4.77-4.72(m,2H,OCH2Ph, H-1 '), 4.68 (q, J=11.4Hz, 2H, OCH2), Ph 4.63 (d, J=12.8Hz, 1H, OCH2Ph),4.59-4.44(m,8H,OCH2Ph,H-2’),4.43-4.36(m, 3H,OCH2Ph,OCH 2CH=CH2, H-1), 4.23 (d, J=2.7Hz, 1H, H-2), 4.02-3.95 (m, 3H, OCH 2CH=CH2, H-6a ', H-4), 3.91 (t, J=9.0Hz, 1H, H-3 "), and 3.88-3.84 (m, 1H, H-6b "), 3.79-3.58 (m, 7H, H- 6a’,H-6b’,H-6a,H-6b,H-5’,H-4”,H-4’),3.54-3.45(m,3H,H-3’,H-3,H-5”),3.37-3.33 (m,1H,H-5),1.96(s,3H),1.02(s,9H).13C NMR(150MHz,CDCl3)δ170.6,138.6,138.5, 138.3,138.25,138.22,138.2,138.1,138.0,135.7,135.5,133.9,133.5,133.1,116.8, 102.2,100.8,100.1,83.9,80.4,79.7,77.9,76.6,75.4,75.2,74.9,74.7,74.6,74.4, 73.5,73.2,69.7,69.6,69.2,68.9,63.9,26.9,21.2,19.2.HRMS(ESI):calcd.for C95H104O17Si[M+Na]+m/z,1567.6942；found,1567.6924.
The synthesis of embodiment 9 compound 12c
Prepared by compound 10 and compound 11, concrete operation step is with the synthesis of compound 12a, yield 81.5%.
1H NMR(600MHz,CDCl3)δ7.46-6.74(m,65H,Ar),5.79-5.73(m,1H,OCH2CH=CH2), 5.56 (d, J=7.8Hz, 1H, H-1 " '), 5.23 (t, J=9.0,7.8Hz, 1H, H-2 " '), 5.16 (s, 1H, H-1 "), 5.16- 5.09(m,3H,OCH2CH=CH 2,H-1”,H-1’),5.06(m,1H,OCH2CH=CH 2), 5.02 (d, J=11.4Hz, 2H, OCH2), Ph 4.95 (d, J=12.0Hz, 1H, OCH2), Ph 4.91 (d, J=10.8Hz, 1H, OCH2), Ph 4.85 (d, J= 10.8Hz,1H,OCH2Ph), 4.83 (d, J=3.0Hz, 1H, H-2 "), 4.80-4.73 (m, 3H, OCH2), Ph 4.66 (q, J= 11.4Hz,2H,OCH2Ph),4.57-4.60(m,3H,OCH2Ph,H-2’),4.52-4.36(m,10H,OCH2Ph,H-2), 4.34-4.30(m,1H,OCH 2CH=CH2),4.27-4.24(m,2H,OCH2Ph,H-1),4.01-3.97(m,2H,H-4’,H- 4”’),3.90-3.85(m,2H,OCH 2CH=CH2,H-3”’),3.82-3.60(m,14H),3.56-3.54(m,1H,H-3’), 3.44-3.38(m,2H,H-3,H-5’),3.31-3.28(m,1H,H-5),1.94(s,3H),0.94(s,9H).13C NMR (150MHz,CDCl3)δ170.6,138.67,138.6,138.5,138.4,138.3,138.2,137.9,137.8,137.3, 135.64,135.62,133.5,133.1,132.8,129.4,129.3,128.4,128.3,128.3,128.22,128.20, 128.17,128.11,128.1,127.9,127.88,127.86,127.8,127.7,127.6,127.5,127.49, 127.44,127.40,127.36,127.33,127.2,126.9,117.2,101.8,100.7,100.5,100.4,84.0, 81.90,80.6,77.4,75.8,75.5,75.3,75.2,75.1,75.00,74.9,74.8,74.7,74.5,74.4, 73.65,73.4,73.3,73.1,72.7,71.2,70.6,70.2,69.6,69.4,69.3,69.0,68.1,62.8,26.9, 21.2,19.2.HRMS(ESI):calcd.for C122H132O22Si[M+Na]+m/z,1999.8877；found,1999.8931.
The synthesis of embodiment 10 compound 13a
Compound 12a (470mg, 4.22mmol) is used 15mL absolute methanol to dissolve, addition sodium methoxide (4.5mg, 0.84mmol) juxtaposition is stirred overnight at room temperature, adds ion exchange resin IR 120 (H+form) neutralization, mistake after completion of the reaction Filter resin, after concentration gained head product through silica gel column chromatography (n-hexane/acetone, 14:1, v/v) obtain white foam solid (417.5g, 92.3%).
1H NMR(600MHz,CDCl3)δ7.77-7.06(m,35H,Ar),6.02-5.95(m,1H,OCH2CH=CH2), 5.39(m,1H,OCH2CH=CH 2),5.28(m,1H,OCH2CH=CH 2), 5.13 (dd, J=11.4Hz, 1H, OCH2Ph),4.99 (d, J=10.8Hz, 1H, OCH2), Ph 4.95 (d, J=12.0Hz, 1H, OCH2Ph), 4.90 (d, J=12.0Hz, 1H, OCH2Ph), 4.88 (d, J=7.6Hz, 1H, H-1 '), 4.85 (d, J=11.4Hz, 1H, OCH2Ph), 4.71 (d, J=12.0Hz, 1H,OCH2Ph),4.62-4.50(m,6H,OCH2Ph,OCH 2CH=CH2, H-1), 4.43 (d, J=3.6Hz, 1H, H-2), 4.15-4.10(m,1H,OCH 2CH=CH2), 4.06 (d, J=10.8Hz, 1H, H-6a '), 4.02 (t, J=9.6Hz, 1H, H- 4),3.96-3.92(m,1H,H-6b’),3.86-3.80(m,3H,H-6a,H-6b,H-2’),3.76-3.71(m,1H,H-3’), 3.65-3.52(m,3H,H-3,H-4’,H-5’),3.50-3.46(m,1H,H-5),1.09(s,9H).13C NMR(150MHz, CDCl3)δ139.2,138.5,138.25,138.21,137.8,135.7,135.6,133.57,133.51,133.5, 129.64,129.60,128.4,128.34,128.29,128.1,128.0,127.99,127.8,127.73,127.70, 127.6,127.57,127.5,117.8,103.7,99.6,85.2,80.3,76.9,75.7,75.2,74.9,74.8,74.3, 73.9,73.5,70.1,70.0,69.1,63.9,26.9,19.3.HRMS(ESI):calcd.for C66H74O11Si[M+Na]+ m/z,1093.4898；found,1093.4884.
The synthesis of embodiment 11 compound 13b
By compound 12b and sodium methoxide preparation, concrete operation step is with the synthesis of compound 13a, yield 95.5%.
1H NMR(600MHz,CDCl3)δ7.71-6.93(m,50H,Ar),5.92-5.86(m,1H,OCH2CH=CH2), 5.22(m,1H,OCH2CH=CH 2),5.16(m,1H,OCH2CH=CH 2), 5.04 (d, J=11.4Hz, 1H, OCH2Ph),4.95 (d, J=10.8Hz, 1H, OCH2Ph),4.98-4.91(m,4H,OCH2Ph, H-1 '), 4.84 (d, J=7.8Hz, 1H, H-1 "), 4.78 (d, J=10.8Hz, 1H, OCH2), Ph 4.63 (d, J=12.6Hz, 1H, OCH2Ph), 4.57 (d, J=11.4Hz, 2H, OCH2Ph), 4.54 (d, J=3.0Hz, 1H, H-2 '), 4.51 (d, J=3.0Hz, 1H, H-2), 4.49-4.35 (m, 9H, OCH2Ph,OCH 2CH=CH2,H-1),4.12-4.02(m,3H,OCH 2CH=CH2,H-6a”,H-4),3.93-3.87(m,1H,H- 6b”),3.83-3.72(m,5H,H-2”,H-4’,H-6a,H-6b,H-6a’),3.71-3.64(m,2H,H-6a’,H-3”), 3.62-3.58(m,1H,H-5’),3.57-3.39(m,5H,H-3’,H-3,H-5’,H-4”,H-5),0.99(s,9H).13C NMR (150MHz,CDCl3)δ139.1,138.57,138.54,138.1,137.9,135.6,133.7,133.5,133.4,129.6, 128.8,128.4,128.3,128.29,128.23,128.22,128.17,128.13,128.01,128.0,127.77, 127.72,127.7,127.56,127.52,127.4,127.3,127.2,117.3,105.1,100.03,100.01,86.6, 80.2,79.5,75.5,75.4,75.2,74.9,74.8,74.6,74.5,74.1,73.5,73.4,71.0,70.2,70.1, 69.6,69.5,64.5,27.0,19.2.HRMS(ESI):calcd.for C93H102O16Si[M+Na]+m/z,1525.6835； found,1525.6777.
The synthesis of embodiment 12 compound 13c
By compound 12c and sodium methoxide preparation, concrete operation step is with the synthesis of compound 13a, yield 91.3%.
1H NMR(600MHz,CDCl3)δ7.64-6.86(m,65H,Ar),5.83-5.76(m,1H,OCH2CH=CH2), 5.31(s,1H,H-1”),5.15(m,1H,OCH2CH=CH 2),5.09(m,1H,OCH2CH=CH 2), 5.06 (d, J= 11.4Hz,1H,OCH2), Ph 5.01 (d, J=10.8Hz, 1H, OCH2Ph), 4.98 (s, 1H, H-1 '), 4.97 (d, J= 10.8Hz,1H,OCH2), Ph 4.90 (d, J=11.4Hz, 1H, OCH2Ph), 4.85 (d, J=3.0Hz, 1H, H-2 '), 4.84- 4.78(m,3H,OCH2Ph), 4.72 (d, J=7.8Hz, 1H, H-1 " '), 4.62 (d, J=12.0Hz, 1H, OCH2Ph),4.59 (d, J=11.4Hz, 2H, OCH2Ph),4.52-4.39(m,10H,OCH2Ph,H-2’),4.37-4.32(m,3H,OCH2Ph, OCH 2CH=CH2, H-1), 4.22 (d, J=12.0Hz, 1H, OCH2), Ph 4.18 (d, J=10.8Hz, 1H, OCH2Ph),4.02 (t, J=9.6Hz, 1H, H-4 "), 3.95-3.88 (m, 4H, OCH 2CH=CH2,H-4’,H-6a”’,H-6b”’),3.85-3.80 (m,2H,H-6a”,H-6b”),3.75-3.55(m,10H),3.54-3.46(m,4H,H-3,H-2,H-5”’,H-5”),3.33- 3.30(m,1H,H-5),0.96(s,9H).13C NMR(150MHz,CDCl3)δ139.1,138.8,138.6,138.5, 138.34,138.30,138.2,138.1,138.0,135.7,135.6,133.7,133.5,133.3,129.5,128.4, 128.4,128.3,128.26,128.24,128.2,128.14,128.09,128.0,127.97,127.93,127.8, 127.77,127.73,127.6,127.5,127.49,127.4,127.36,127.3,127.2,127.0,117.2,105.7, 101.4,100.3,99.7,86.2,81.0,80.3,79.8,76.6,75.8,75.7,75.6,75.5,75.4,75.34, 75.32,75.2,74.8,74.7,74.5,73.4,72.2,70.2,70.1,70.0,69.99,69.91,69.7,69.4, 69.2,63.9,26.9,19.3.HRMS(ESI):calcd.for C120H130O21Si[M+Na]+m/z,1957.8772；found, 1957.8856.
The synthesis of embodiment 13 compound 14a
Compound 13a (380.0mg, 0.36mmol) is placed in 25mL round-bottomed flask, is subsequently adding dimethyl sulfoxide (DMSO) (5.0mL) with aceticanhydride (2.5mL).Reactant liquor is extracted with ethyl acetate after being stirred at room temperature 18 hours, uses saturated carbon respectively Sour sodium and brine It, then use anhydrous sodium sulfate drying, and the head product being concentrated to give after filtration toluene band revolves twice.Residual Stay thing 20mL anhydrous tetrahydro furan to dissolve and be cooled to -78 DEG C, under argon gas protection, be slowly added dropwise L-Selectride (1M THF, 1.75mL) and stir 15 minutes, then remove cooling device juxtaposition and continue at room temperature to stir 15 minutes, methyl alcohol is quenched Go out and after reacting, add 20mL dchloromethane, solution uses hydrogenperoxide steam generator (5%, 20mL), sodium hydroxide solution respectively (1M, 20mL), hypo solution (5%, 20mL) and saturated aqueous common salt (20mL) washing, through anhydrous magnesium sulfate be dried, mistake Filter and concentrate after pale yellow oily liquid, head product through silica gel column chromatography (n-hexane/ethyl acetate, 6:1, v/v) must be white Foaming solid (323.5mg, 84.5%).
1H NMR(600MHz,CDCl3)δ7.69-7.07(m,35H,Ar),5.92-5.84(m,1H,OCH2CH=CH2), 5.26(m,1H,OCH2CH=CH 2),5.16(m,1H,OCH2CH=CH 2),5.12(br s,1H,H-1’),4.96-4.90(m, 2H,OCH2Ph),4.88-4.82(m,2H,OCH2Ph), 4.69 (d, J=3.6Hz, 1H, H-2), 4.67-4.62 (m, 2H, OCH2Ph) 4.57 (d, J=12.0Hz, 1H, OCH2Ph),4.51-4.37(m,5H,OCH2Ph,OCH 2CH=CH2,H-1),4.36 (d, J=3.0Hz, 1H, H-2 '), 4.09-4.03 (m, 2H, OCH 2CH=CH2,H-6a’),3.91-3.79(m,4H,H-6b’,H- 4,H-4’,H-6a),3.76-3.72(m,1H,H-6b),3.64-3.57(m,2H,H-3’,H-3),3.51-3.43(m,2H,H- 5’,H-5),1.03(s,9H).13C NMR(150MHz,CDCl3)δ139.1,138.6,138.4,138.24,138.20, 133.9,128.8,128.49,128.42,128.4,128.2,128.12,128.10,127.8,127.76,127.72, 127.6,127.5,127.4,117.2,102.3,100.1,81.7,80.7,75.9,75.5,75.24,75.20,74.46, 74.42,74.3,73.8,73.6,73.4,70.8,70.4,70.0,69.3,62.5.HRMS(ESI):calcd.for C66H74O11Si[M+Na]+m/z,1093.4898；found,1093.4882.
The synthesis of embodiment 14 compound 14b
Prepared through aceticanhydride/dimethylsulfoxide oxidation, L-Selectride reduction reaction by compound 13b, concrete operation step With the synthesis of compound 14a, yield 81.5%.
1H NMR(600MHz,CDCl3)δ7.65-6.89(m,50H,Ar),5.88-5.82(m,1H,OCH2CH=CH2), 5.29(s,1H,H-1”),5.20(m,1H,OCH2CH=CH 2),5.16(m,1H,OCH2CH=CH 2),5.12(s,1H,H-1’), 4.94 (d, J=10.8Hz, 2H, OCH2), Ph 4.90 (d, J=12.0Hz, 1H, OCH2Ph), 4.85 (d, J=10.8Hz, 1H, OCH2), Ph 4.77 (d, J=10.8Hz, 1H, OCH2Ph), 4.73 (d, J=3.0Hz, 1H, H-2 '), 4.63 (d, J=3.0Hz, 1H,H-2),4.51-4.42(m,6H,OCH2Ph,H-1),4.41-4.37(m,3H,OCH2Ph,OCH 2CH=CH2),4.34(d,J =10.8Hz, 1H, OCH2Ph), 4.32 (d, J=2.4Hz, 1H, H-2 "), 4.22 (d, J=10.8Hz, 2H, OCH2Ph),4.06- 3.97(m,3H,OCH2Ph,OCH 2CH=CH2,H-6a”),3.92-3.86(m,2H,H-4”,H-4’),3.80-3.72(m,2H, H-6a’,H-6b’),3.69-3.55(m,5H,H-6a,H-6b,H-4,H-3’,H-3),3.54-3.47(m,3H,H-5”,H-5’, H-3”),3.41-3.37(m,1H,H-5).13C NMR(150MHz,CDCl3)δ138.5,138.4,138.2,138.1,138.1, 138.07,138.03,137.9,135.63,135.61,133.6,133.5,133.4,129.5,128.9,128.5,128.3, 128.3,128.2,128.2,128.1,128.0,127.9,127.85,127.82,127.7,127.69,127.65,127.63, 127.5,127.4,127.4,127.2,127.1,117.3,100.6,100.3,99.6,83.1,80.4,79.9,76.6, 75.4,75.3,75.1,74.8,74.8,74.5,74.1,73.4,70.8,70.3,70.2,70.1,69.9,69.6,69.3, 69.0,67.5,64.1,26.9,19.3.HRMS(ESI):calcd.for C93H102O16Si[M+Na]+m/z,1525.6835； found,1525.6805.
The synthesis of embodiment 15 compound 14c
Prepared through aceticanhydride/dimethylsulfoxide oxidation, L-Selectride reduction reaction by compound 13c, concrete operation step With the synthesis of compound 14a, yield 79.6%.
1H NMR(600MHz,CDCl3)δ7.69-6.95(m,65H,Ar),5.85-5.78(m,1H,OCH2CH=CH2), 5.47(s,1H,H-1”’),5.26(s,1H,H-1”),5.19(m,1H,OCH2CH=CH 2),5.11(m,1H,OCH2CH= CH 2), 5.01 (s, 1H, H-1 '), 4.98 (d, J=10.8Hz, 1H, OCH2Ph),4.94-4.91(m,2H,OCH2Ph,H-2’), 4.94-4.91(m,4H,OCH2), Ph 4.68 (d, J=12.0Hz, 1H, OCH2Ph),4.61-4.57(m,3H,OCH2Ph,H- 2”’),4.50-4.39(m,7H,OCH2Ph), 4.38 (d, J=3.0Hz, 1H, H-2 "), 4.36-4.31 (m, 4H, OCH2Ph,H- 2,H-1,OCH 2CH=CH2),4.24-4.21(m,3H,OCH2Ph),4.09-4.01(m,5H,OCH2Ph,H-6a”’,H-6b”’, H-4”’,H-4”),3.95-3.92(m,1H,OCH 2CH=CH2),3.62-3.60(m,10H),3.56-3.49(m,5H,H-3”, H-6a,H-5’,H-5”,H-3),3.35-3.32(m,1H,H-5),1.03(s,9H).13CNMR(150MHz,CDCl3)δ138.9, 138.69,138.49,138.4,138.24,138.20,138.1,138.04,138.01,138.0,135.7,135.6, 133.7,133.6,133.4,129.48,129.5,129.0,128.4,128.33,128.32,128.30,128.27, 128.22,128.20,128.1,128.0,127.9,127.8,127.79,127.76,127.73,127.64,127.62, 127.5,127.45,127.42,127.3,127.2,127.1,127.0,117.1,101.8,100.4,100.2,99.8, 83.1,80.7,80.6,79.7,76.6,75.5,75.44,75.3,75.2,75.1,74.8,74.7,73.9,73.5,73.4, 72.2,72.0,70.3,70.0,69.9,69.88,69.8,69.7,69.1,69.0,68.7,67.6,63.9,26.9, 19.3.HRMS(ESI):calcd.for C120H130O21Si[M+Na]+m/z,1957.8772；found,1957.8750.
The synthesis of embodiment 16 compound 15a
Compound 14a (85.0mg, 0.08mmol) is dissolved in 25mL N ' dinethylformamide and adds tetrabutyl iodate Ammonium (1.0mg, 0.003mmol) and cylite (19 μ L, 0.16mmol), reactant liquor be cooled to 0 DEG C after add sodium hydride (4.8mg, 0.12mmol) and continue reaction one hour under argon gas protection, methyl alcohol adds 50mL dchloromethane, then after reaction is quenched With 30mL saturated common salt water washing, organic layer be dried through anhydrous magnesium sulfate, filter and concentrate after head product, head product is through silica gel Column chromatography (n-hexane/acetone, 18:1, v/v) obtain white foam solid (77.0mg, 83.7%)
1H NMR(600MHz,CDCl3)δ7.79-7.10(m,40H,Ar),6.02-5.93(m,1H,OCH2CH=CH2), 5.36(m,1H,OCH2CH=CH 2),5.25(m,1H,OCH2CH=CH 2), 5.24 (br s, 1H, H-1 '), 5.21 (d, J= 10.8Hz,1H,OCH2Ph),5.03-4.91(m,3H,OCH2Ph), 4.79 (d, J=3.0Hz, 1H, H-2), 4.71-4.58 (m, 4H,OCH2Ph,H-1),4.57-4.53(m,1H,OCH 2CH=CH2),4.52-4.44(m,1H,OCH2), Ph 4.30 (d, J= 3.0Hz,1H,H-2’),4.18-4.13(m,2H,OCH 2CH=CH2,H-6a’),4.07-4.03(m,1H,OCH 2CH=CH2,H- 6b '), 3.95-3.84 (m, 3H, H-4, H-4 ', H-6a), 3.80-3.76 (m, 1H, H-6b), 3.73 (d, J=9.2,3.6Hz, 1H, H-3), 3.67 (d, J=9.2,3.2Hz, 1H, H-3 '), and 3.65-3.60 (m, 1H, H-5 '), 3.58-3.54 (m, 1H, H- 5),1.12(s,9H).13C NMR(150MHz,CDCl3)δ139.5,138.53,138.51,138.5,138.4,135.7, 135.6,133.9,133.7,133.5,129.6,129.58,128.7,128.6,128.37,128.35,128.3,128.2, 128.04,128.02,127.9,127.72,127.70,127.6,127.5,127.4,127.1,117.1,100.3,100.8, 82.3,80.5,77.4,75.6,75.2,75.1,75.0,74.2,74.1,73.7,73.5,70.7,70.6,70.2,69.6, 69.5,64.4,26.9,19.3.HRMS(ESI):calcd.forC73H80O11Si[M+Na]+m/z,1183.5368；found, 1183.5321.
The synthesis of embodiment 17 compound 15b
By compound 14b and cylite reaction preparation, concrete operation step is with the synthesis of compound 15a, yield 89.4%.
1H NMR(600MHz,CDCl3)δ7.69-6.93(m,55H,Ar),5.93-5.85(m,1H,OCH2CH=CH2), 5.41(s,1H,H-1”),5.24(m,1H,OCH2CH=CH 2),5.21-5.15(m,3H,OCH2CH=CH 2,OCH2Ph,H-1’), 4.94 (d, J=11.4Hz, 1H, OCH2), Ph 4.96 (d, J=10.8Hz, 1H, OCH2Ph),4.90-4.84(m,4H, OCH2Ph),4.81-4.75(m,2H,OCH2Ph, H-2 '), 4.65 (d, J=3.0Hz, 1H, H-2), 4.57 (d, J=11.4Hz, 1H,OCH2), Ph 4.52 (d, J=12.0Hz, 1H, OCH2Ph),4.47-4.30(m,9H,H-1,OCH2Ph),4.20(br s, 1H,H-2”),4.17-4.12(m,1H,OCH2Ph),4.08-3.98(m,4H,H-6a”,H-6b”,OCH 2CH=CH2, OCH2Ph), 3.91 (t, J=10.8,9.6Hz, 1H, H-4 "), 3.84-3.76 (m, 2H, H-4 ', H-6a), 3.73-3.66 (m, 4H,H-4,H-6b,H-6a’,H-6b’),3.65-3.57(m,4H,H-3’,H-3,H-3”,H-5”),3.56-3.51(m,1H,H- 5’),3.45-3.41(m,1H,H-5),1.01(s,9H).13C NMR(150MHz,CDCl3)δ140.0,138.8,138.6, 138.53,138.50,138.3,138.2,138.1,137.9,135.7,133.8,133.7,133.6,129.4,128.6, 128.4,128.3,128.24,128.20,128.17,128.15,128.1,127.99,127.95,127.8,127.7, 127.66,127.60,127.56,127.54,127.51,127.46,127.3,127.2,127.0,126.8,117.2, 101.5,100.7,83.4,80.6,80.2,75.37,75.34,75.2,75.1,74.9,74.8,74.8,74.6,74.4, 74.3,73.4,73.3,70.8,70.5,70.2,70.18,69.9,69.7,69.1,68.9,64.4,26.9,19.3.HRMS (ESI):calcd.for C100H108O16Si[M+Na]+m/z,1615.7340；found,1615.7274.
The synthesis of embodiment 18 compound 15c
By compound 14c and cylite reaction preparation, concrete operation step is with the synthesis of compound 15a, yield 87.9%.
1H NMR(600MHz,CDCl3)δ7.69-6.97(m,70H,Ar),5.85-5.78(m,1H,OCH2CH=CH2), 5.49(s,1H,H-1”’),5.40(s,1H,H-1”),5.25(m,1H,OCH2CH=CH 2),5.18(m,1H,OCH2CH= CH 2), 5.11 (d, J=10.8Hz, 1H, OCH2Ph), 5.03 (s, 1H, H-1 '), 4.99 (d, J=11.4Hz, 1H, OCH2Ph), 4.94-4.91(m,2H,OCH2Ph,H-2’),4.87-4.77(m,6H,OCH2Ph), 4.65 (d, J=3.0Hz, 1H, H-2 " '), 4.59 (d, J=12.0Hz, 1H, OCH2Ph),4.54-4.34(m,13H,OCH2Ph,OCH 2CH=CH2,H-2,H-1),4.32 (d, J=10.8Hz, 1H, OCH2Ph), 4.26 (d, J=3.0Hz, 1H, H-2 "), 4.24 (d, J=10.8Hz, 1H, OCH2Ph), 4.13-4.17(m,2H,OCH2Ph),4.10-4.04(m,4H,OCH2Ph,H-6a”’,H-6b”’,H-4”),3.97-3.92(m, 1H,OCH2Ph), 3.86 (t, J=9.6Hz, 1H, H-4 '), 3.80-3.59 (m, 11H), 3.58-3.48 (m, 4H, H-6a, H- 5”’,H-5”,H-3),3.37-3.33(m,1H,H-5),0.98(s,9H).13C NMR(150MHz,CDCl3)δ140.3, 138.9,138.9,138.7,138.6,138.4,138.3,138.2,138.1,138.0,137.98,137.92,135.74, 135.70,133.8,133.7,133.4,129.4,129.3,128.7,128.5,128.38,128.4,128.3,128.29, 128.27,128.24,128.20,128.0,127.96,127.92,127.90,127.87,127.86,127.77,127.73, 127.7,127.62,127.60,127.56,127.50,127.4,127.3,127.2,127.1,127.0,126.9,126.7, 117.2,101.8,100.4,100.3,83.5,80.9,80.7,79.5,75.4,75.39,75.33,75.2,75.0,74.9, 74.8,74.7,74.5,74.4,73.4,73.39,73.36,71.9,70.4,70.0,69.9,69.7,69.7,69.1,68.9, 68.7,64.0,26.9,19.3.HRMS(ESI):calcd.for C127H136O21Si[M+Na]+m/z,2047.9241；found, 2047.9266.
The synthesis of embodiment 19 compound 16a
Compound 15a (183.0mg, 0.16mmol) is dissolved in 5mL oxolane, is subsequently adding tetrabutyl ammonium fluoride (0.63mL,1M), stirring under room temperature added 30mL dchloromethane after one day, then uses 30mL saturated common salt water washing, has Machine layer through anhydrous magnesium sulfate be dried, filter and concentrate after head product, head product through silica gel column chromatography (n-hexane/acetone, 10: 1, v/v) obtain white foam solid (77.0mg, 83.4%)
1H NMR(600MHz,CDCl3)δ7.63-7.15(m,30H,Ar),5.97-5.88(m,1H,OCH2CH=CH2), 5.28(m,1H,OCH2CH=CH 2),5.21(m,1H,OCH2CH=CH 2), 5.15 (d, J=12.0Hz, 1H, OCH2Ph),5.04- 4.96(m,5H,OCH2Ph, H-1 '), 4.93 (d, J=10.8Hz, 1H, OCH2Ph), 4.71 (d, J=10.8Hz, 1H, OCH2Ph),4.66-4.60(m,2H,OCH2Ph),4.58-4.47(m,5H,OCH2Ph, H-1), 4.43 (d, J=12.0Hz, 1H, OCH2), Ph 4.39 (d, J=3.0Hz, 1H, H-2), 4.29 (d, J=3.0Hz, 1H, H-2), 4.10-4.07 (m, 1H, OCH 2CH =CH2), 4.06 (t, J=9.6Hz, 1H, H-4 '), 3.95 (t, J=9.6Hz, 1H, H-4), 3.92-3.84 (m, 2H, H-6a ', H-6b '), 3.82 (d, J=10.8Hz, 1H, H-6a), 3.76 (dd, J=10.8,5.0Hz, 1H, H-6b), 3.66 (dd, J= 9.3,2.9Hz, 1H, H-3), 3.59 (dd, J=9.6,3.0Hz, 1H, H-3 '), 3.52-3.47 (m, 1H, H-5), 3.44-3.41 (m,1H,H-5’),13C NMR(150MHz,CDCl3)δ139.10,138.56,138.36,138.24,138.20,133.89, 128.78,128.49,128.42,128.35,128.18,128.12,128.10,127.83,127.76,127.72,127.61, 127.52,127.37,117.20,102.25,100.15,81.69,80.68,77.42,77.21,77.00,75.98,75.46, 75.24,75.20,74.46,74.42,74.33,73.80,73.56,73.43,70.81,70.42,70.00,69.30, 62.49.HRMS(ESI):calcd.forC57H62O11[M+Na]+m/z,945.4190；found,945.4161.
The synthesis of embodiment 20 compound 16b
By compound 15b and tetrabutyl ammonium fluoride reaction preparation, concrete operation step is with the synthesis of compound 16a, yield 88.2%.
1H NMR(600MHz,CDCl3)δ7.51-6.95(m,45H,Ar),5.92-5.84(m,1H,OCH2CH=CH2), 5.29(s,1H,H-1”),5.24(m,1H,OCH2CH=CH 2),5.19-5.15(m,2H,OCH2CH=CH 2,H-1’),5.07 (d, J=12.0Hz, 1H, OCH2), Ph 4.98 (d, J=11.4Hz, 1H, OCH2Ph), 4.93 (d, J=10.8Hz, 1H, OCH2Ph),4.89-4.84(m,3H,OCH2), Ph 4.71 (d, J=12.0Hz, 1H, OCH2Ph), 4.71 (d, J=3.0Hz, 1H, H-2 '), 4.65 (d, J=12.0Hz, 1H, OCH2), Ph 4.61 (d, J=3.0Hz, 1H, H-2), 4.56 (d, J=10.8Hz, 1H,OCH2Ph),4.46-4.40(m,6H,OCH2Ph,H-1,OCH 2CH=CH2), 4.29 (d, J=10.8Hz, 1H, OCH2Ph), 4.15 (d, J=3.0Hz, 1H, H-2 "), 4.13 (d, J=11.4Hz, 1H, OCH2Ph), 4.07 (d, J=11.4Hz, 1H, OCH2Ph),4.03-3.98(m,1H,OCH 2CH=CH2), 3.94 (t, J=9.6Hz, 1H, H-4 "), 3.86 (t, J=9.6Hz, 1H,H-4’),3.76-3.80(m,2H,H-6a”,H-6a’),3.75-3.67(m,5H,H-6b”,H-4,H-6a,H-6b,H- 6b’),3.66-3.63(m,1H,H-3’),3.63-3.58(m,1H,H-3),3.55-3.51(m,2H,H-3”,H-5’),3.42- 3.35(m,2H,H-5,H-5”).13C NMR(150MHz,CDCl3)δ139.6,138.7,138.5,138.4,138.4,138.2, 138.1,138.0,137.7,133.6,128.6,128.41,128.40,128.4,128.3,128.29,128.23,128.2, 128.04,128.00,127.8,127.7,127.69,127.65,127.61,127.6,127.4,127.1,127.1,117.4, 101.6,100.7,100.4,83.0,80.7,80.6,75.5,75.4,75.2,75.1,74.7,74.66,74.62,74.6, 74.5,73.6,73.4,71.6,70.5,70.4,69.9,69.8,69.6,68.7,62.4.HRMS(ESI):calcd.for C84H90O16[M+Na]+m/z,1377.6127；found,1377.6112.
The synthesis of embodiment 21 compound 16c
By compound 15c and tetrabutyl ammonium fluoride reaction preparation, concrete operation step is with the synthesis of compound 16a, yield 87.2%.
1H NMR(500MHz,CDCl3)δ7.52-6.02(m,60H,Ar),5.92-5.83(m,1H,OCH2CH=CH2), 5.53(s,1H,H-1”’),5.32(s,1H,H-1”),5.24(m,1H,OCH2CH=CH 2),5.18(m,1H,OCH2CH= CH 2), 5.15 (s, 1H, H-1 '), 4.99 (d, J=12.0Hz, 1H, OCH2Ph),5.00-4.88(m,5H,OCH2Ph,H-2’), 4.83 (d, J=10.8Hz, 1H, OCH2), Ph 4.75 (d, J=12.0Hz, 1H, OCH2Ph),4.87-4.77(m,15H, OCH2Ph,H-2,H-2”’,OCH 2CH=CH2), 4.25 (d, J=3.0Hz, 1H, H-2 "), 4.18-4.10 (m, 2H, OCH2Ph), 4.05-3.96(m,3H,OCH 2CH=CH2,OCH2Ph),3.87-3.54(m,15H),3.48-3.40(m,2H,H-5,H-5”) .13C NMR(125MHz,CDCl3)δ139.6,138.75,138.72,138.6,138.5,138.4,138.2,138.16, 138.1,137.9,137.8,133.6,128.62,128.60,128.5,128.45,128.4,128.3,128.32,128.3, 128.25,128.22,128.21,128.2,128.1,128.04,128.02,128.0,127.8,127.72,127.7, 127.6,127.5,127.4,127.3,127.1,127.0,117.3,101.7,101.2,100.7,100.2,82.9,81.8, 80.7,80.4,75.6,75.5,75.3,75.2,75.1,75.0,74.8,74.8,74.5,74.4,73.6,73.5,73.45, 72.6,70.5,70.5,70.3,70.1,69.8,69.6,69.6,69.5,69.3,68.8,62.4.MALDI-TOF-MS: calcd.for C111H118O21[M+Na]+m/z,1811.10；found,1810.76.
The synthesis of embodiment 22 compound 18a
By compound 16a (50.0mg, 0.06mmol) and phosphorylation agent 17 (138mg, 0.26mmol) (Synthesis.1992,1269.) is placed in 10mL round-bottomed flask and adds 3.0mL dichloromethane to dissolve, after solution is cooled to 0 DEG C Dropping 1H-TETRAZOLE solution (0.6mL, 0.45M), the lower reaction of argon gas protection is removed cooling device after 15 minutes and is continued at room temperature Stirring two hours, after being subsequently cooled to -40 DEG C dropping tert-Butanol peroxide (0.07mL, 5.5M) solution, remove cooling device and continue After stirring three hours under room temperature, add 15mL 10% hypo solution that reaction is quenched, reactant liquor is extracted with 30mL dichloromethane Take, then washed with saturated sodium bicarbonate solution (30mL) and saline solution (30mL) successively, organic layer through anhydrous magnesium sulfate be dried, Filter and concentrate after head product, head product through silica gel column chromatography (n-hexane/ethyl acetate, 3:2, v/v) obtain diastereo-isomerism Mixture (66.7mg, 82.0%).
Selected signals:1H NMR(500MHz,CDCl3)δ4.97(br s,1H),4.41(br s,1H).13C NMR(125MHz,CDCl3)δ102.14,102.09,100.31,100.26.31P NMR(160MHz,CDCl3)δ-0.61,- 0.76.HRMS(ESI):calcd.for C81H84NO16P[M+Na]+m/z,1380.5425；found,1380.5381.
The synthesis of embodiment 23 compound 18b
Preparation is reacted by compound 16b and phosphorylation agent 17, concrete operation step is with the synthesis of compound 18a, yield 87.5%.
Selected signals:1HNMR(500MHz,CDCl3)δ5.40(br s,1H),5.18(br s,1H),4.46 (br s,1H).13C NMR(125MHz,CDCl3)δ101.62,101.52,100.79,100.70,100.65,100.54.31P NMR(160MHz,CDCl3)δ-0.50,-0.70.HRMS(ESI):calcd.for C108H112NO21P[M+Na]+m/z, 1812.7362；found,1812.7319.
The synthesis of embodiment 24 compound 18c
Preparation is reacted by compound 16c and phosphorylation agent 17, concrete operation step is with the synthesis of compound 18a, yield 79.3%.
NMR signals for 18c(isomer 1):1H NMR(600MHz,CDCl3)δ7.73-6.93(m,73H, Ar),5.85-5.78(m,1H,OCH2CH=CH2),5.48(s,1H,H-1”’),5.30(s,1H,H-1”),5.19(m,1H, OCH2CH=CH 2),5.14-5.10(m,1H,OCH2Ph),5.09(s,1H,H-1’),5.05-5.00(m,1H,OCH2Ph), 4.97-4.82(m,9H,OCH2Ph, H-2 '), 4.71 (d, J=12.0Hz, 1H, OCH2Ph), 4.62 (d, J=12.0Hz, 1H, OCH2), Ph 4.59 (d, J=12.0 Hz, 1H, OCH2), Ph 4.56 (d, J=10.8 Hz, 1H, OCH2), Ph 4.57 (d, J= 12.0 Hz,1H,OCH2Ph),4.53-4.44(m,6H,OCH2Ph, H-1, H-2 " '), 4.41-4.32 (m, 8H), 4.29 (d, J= 3.6 Hz, 1H, H-2 "), 4.26 (d, J=10.8 Hz, 1H, OCH2Ph),4.15-4.06(m,4H),4.03-3.93(m,4H), 3.92-3.75 (m, 5H), 3.72-3.58 (m, 8H), 3.55 (dd, J=9.6,3.6 Hz, 1H, OCH2Ph),3.52-3.43(m, 2H),3.37-3.32(m,1H),3.26-3.18(m,1H),3.08-2.99(m,1H).13C NMR(150 MHz,CDCl3)δ 156.18,144.08,143.92,141.16,141.12,139.49,138.64,138.52,138.43,138.34,138.21, 138.05,138.02,137.99,137.84,137.75,135.80,136.76,133.59,128.64,128.57,128.52, 128.45,128.32,128.28,128.26,128.22,128.18,128.16,128.12,128.04,128.02,127.97, 127.96,127.88,127.85,127.81,127.70,127.64,127.56,127.54,127.50,127.43,127.39, 127.24,127.11,127.06,126.97,126.95,125.21,125.12,119.77,117.31,102.08,101.31, 100.55,100.29,83.10,81.97,80.53,80.35,75.52,75.31,75.27,75.12,75.05,74.69, 74.58,74.24,74.20,73.61,73.56,73.41,73.21,70.92,70.54,70.19,70.18,69.69, 69.60,69.51,69.36,69.15,69.12,68.77,66.89,66.85,66.57,66.45,47.04,40.95.31P NMR(160 MHz,CDCl3)δ-0.67.MALDI-TOF-MS:calcd.for C108H112NO21P[M+H]+m/z,2222.947； found,2223.236.NMR signals for 18c(isomer 2):1H NMR(600 MHz,CDCl3)δ7.73-6.93 (m,73H,Ar),5.87-5.78(m,1H,OCH2CH=CH2),5.48(s,1H,H-1”’),5.32(s,1H,H-1”),5.19 (m,1H,OCH2CH=CH 2),5.15-5.11(m,2H,OCH2Ph),5.07(s,1H,H-1’),5.06-5.00(m,3H, OCH2Ph),4.97-4.82(m,8H,OCH2Ph, H-2 '), 4.74 (d, J=11.4 Hz, 1H, OCH2), Ph 4.69 (d, J= 12.0 Hz,1H,OCH2), Ph 4.66 (d, J=12.0 Hz, 1H, OCH2), Ph 4.62 (d, J=10.8 Hz, 1H, OCH2Ph), 4.57 (d, J=12.0 Hz, 1H, OCH2), Ph 4.54 (d, J=12.0 Hz, 1H, OCH2Ph),4.51(br s,1H,H-1), 4.49 (d, J=10.8 Hz, 2H, OCH2Ph),4.47-4.44(m,3H,OCH2Ph,H-2”’),4.41-4.34(m,8H),4.27 (m, 2H), 4.26 (d, J=3.0 Hz, 1H, H-2 "), 4.22 (d, J=10.8 Hz, 2H, OCH2Ph),4.13-4.04(m, 3H), 4.03-3.93 (m, 3H), 3.87-3.83 (m, 3H), 3.68-3.58 (m, 9H), 3.55 (dd, J=9.6,3.6 Hz, 1H, OCH2Ph),3.52-3.47(m,2H),3.37-3.34(m,1H),3.14-3.08(m,2H).13CNMR(150 MHz,CDCl3)δ 156.22,144.01,143.97,141.19,139.59,138.68,138.60,138.33,138.26,138.11,138.08, 138.04,137.88,137.78,136.08,136.04,133.61,128.67,128.52,128.46,128.42,128.37, 128.30,128.29,128.23,128.22,128.17,128.09,128.05,128.01,127.98,127.96,127.92, 127.79,127.72,127.66,127.63,127.58,127.53,127.40,127.21,127.18,127.09,127.05, 127.00,126.57,125.16,119.81,117.27,101.97,101.47,100.52,100.27,83.12,81.73, 80.62,80.31,75.47,75.35,75.28,75.23,75.09,75.01,74.97,74.94,74.70,74.51, 74.30,74.26,73.60,73.53,73.40,73.37,72.86,71.25,70.43,70.15,70.12,69.70, 69.63,69.59,69.57,69.42,69.27,69.24,68.83,66.70,66.66,66.59,47.11,41.16, 41.12.31P NMR(160MHz,CDCl3)δ-0.52.HRMS(ESI):calcd.for C108H112NO21P[M+Na]+m/z, 2244.9293；found,2244.9475.
The synthesis of embodiment 25 compound 19a
Compound 18a (50.0mg, 0.037mmol) is placed in 5mL eggplant type bottle and adds 0.8mL dichloromethane to dissolve, room Stir under temperature and drip 51,8- diazabicylos 11 carbon -7- alkene (DBU), after two minutes, add 5 acetic acid neutralization reactions, Under low-temp low-pressure remove solvent after gained head product through silica gel column chromatography (methylene chloride/methanol, 30:1, v/v) obtain diastereomeric different Structure mixture (38.0mg, 90.4%).
Selected signals:1H NMR(600MHz,CDCl3)δ4.96(br s,1H),4.40(br s,1H).13C NMR(125MHz,CDCl3)δ101.84,101.69,100.14,99.86.HRMS(ESI):calcd.for C66H75NO14P [M+H]+m/z,1136.4920；found,1136.4925.
The synthesis of embodiment 26 compound 19b
Preparation is reacted by compound 18b and DBU, concrete operation step is with the synthesis of compound 19a, yield 93.2%.
Selected signals:1H NMR(600MHz,CDCl3)δ5.31(s,1H),5.13(s,1H),4.44(s, 1H).13CNMR(150MHz,CDCl3)δ101.27,100.78,100.67,100.55,100.22,100,08.HRMS(ESI): calcd.for C93H102NO19P[M-H]-m/z,1566.6711；found,1566.6705.
The synthesis of embodiment 27 compound 19c
Preparation is reacted by compound 18c and DBU, concrete operation step is with the synthesis of compound 19a, yield 86.3%.
NMR signals for 19c(isomer 1):1H NMR(600MHz,CDCl3)δ7.50-6.92(m,65H, Ar),5.85-5.77(m,1H,OCH2CH=CH2),5.46(s,1H,H-1”’),5.31(s,1H,H-1”),5.18(m,1H, OCH2CH=CH 2),5.04(s,1H,H-1’),5.03-4.97(m,5H,OCH2Ph),4.90-4.78(m,5H,OCH2Ph), 4.67-4.60(m,3H),4.59-4.45(m,6H,OCH2Ph),4.43-4.33(m,8H,OCH2Ph,H-1),4.31-4.19(m, 3H), 4.09 (d, J=11.2Hz, 1H, OCH2), Ph 4.04 (d, J=11.2Hz, 1H, OCH2Ph),3.98-3.88(m,3H), 3.84-3.68 (m, 6H), 3.67-3.56 (m, 6H), 3.54 (dd, 1H, J=9.6,3.0Hz), 3.51-3.45 (m, 1H), 3.44-3.40(m,1H),3.37-3.32(m,1H).13C NMR(150MHz,CDCl3)δ138.78,138.49,138.24, 138.16,138.11,138.05,137.99,137.87,137.71,133.59,128.76,128.56,128.50,128.42, 128.40,128.31,128.30,128.24,128.17,128.16,128.11,128.07,128.04,128.00,127.98, 127.90,127.85,127.76,127.73,127.69,127.64,127.51,127.40,127.20,127.15,127.10, 117.25,101.65,101.59,100.40,100.10,83.33,81.50,80.66,80.26,75.46,75.36,75.27, 75.23,75.03,74.99,74.95,74.73,74.59,74.35,73.58,73.51,73.38,70.25,70.08, 69.78,69.52,69.48,69.40,69.03,68.83,66.60.NMRsignals for 19c(isomer 2):1H NMR (600MHz,CDCl3)δ7.55-6.93(m,65H,Ar),5.85-5.79(m,1H,OCH2CH=CH2),5.53(s,1H,H- 1”’),5.38(s,1H,H-1”),5.18(m,1H,OCH2CH=CH 2),5.15-5.11(m,2H,OCH2Ph,H-1’),5.10 (s,1H,H-1’),5.01-4.91(m,5H,OCH2), Ph 4.89 (d, J=3.0Hz, 1H), 4.85-4.78 (m, 3H), 4.66 (d, J=10.8Hz, 1H, OCH2Ph),4.61-4.57(m,2H),4.47-4.37(m,14H,OCH2Ph,H-2,H-1),4.29 (d, J=10.8Hz, 1H, OCH2), Ph 4.25-4.14 (m, 3H), 4.07 (d, J=11.2Hz, 1H, OCH2Ph),3.98-3.75 (m,7H),3.71-3.55(m,9H),3.51-3.44(m,2H),3.39-3.34(m,2H).13C NMR(150MHz,CDCl3)δ 138.94,138.20,138.13,138.05,138.02,138.00,137.98,137.91,137.84,137.71,133.61, 129.19,128.93,128.62,128.56,128.52,128.46,128.37,128.34,128.30,128.25,128.21, 128.07,128.04,128.02,127.95,127.93,127.92,127.87,127.84,127.74,127.67,127.60, 127.55,127.50,127.35,127.24,126.92,117.26,101.32,101.15,100.46,99.78,82.15, 80.61,80.54,75.58,75.39,75.30,75.20,75.09,75.02,74.93,74.82,74.65,74.49, 73.61,73.48,73.43,71.13,70.38,70.30,70.12,70.04,69.33,69.16,69.09,69.05, 68.89,66.31,60.03.HRMS(ESI):calcd.for C120H131NO24P[M+H]+m/z,2000.8793；found, 2000.8799.
The synthesis of embodiment 28 compound 20a
Compound 19a (20.0mg) and hydroxide palladium carbon (10Wt.%, 10.0mg) are placed in 5mL test tube and add first Alcohol/dichloromethane mixed solution (2mL, 1:1,v:V), then test tube is put in hydrogenation reaction cauldron and shake under 40 atmospheric pressure Swing reaction one day, suspension is filtered with diatomite after completion of the reaction, filter cake is washed with methyl alcohol, merge cleaning solution and concentrate, residual Thing obtains white solid (8.2mg, 91.5%) through LH-20 column chromatography after being washed with ethyl acetate.
1H NMR(600MHz,CDCl3)δ4.84(s,1H,H-1’),4.58(s,1H,H-1),4.26-4.21(m,1H,), 4.14-4.07(m,4H,H-2),4.05-4.02(m,1H,H-2’),3.90-3.84(m,2H,OCH 2CH2CH3),3.76-3.68 (m,2H,H-4’),3.57-3.45(m,3H,OCH 2CH2CH3),3.42-3.38(m,1H,H-3’),3.33(s,1H,),3.30- 3.27(m,3H,),3.24-3.12(m,4H,H-5’),1.64-1.57(m,2H,OCH2 CH 2CH3), 0.94 (t, J=7.8Hz, 1H,OCH2CH2 CH 3).13C NMR(150MHz,CDCl3)δ101.33,100.47,78.16,76.91,75.69,73.53, 72.83,70.86,70.61,66.99,66.12,64.48,61.82,61.11,40.05,22.59,9.60.HRMS(ESI): calcd.forC17H33NO14P[M-H]-m/z,506.1639；found,506.1628
The synthesis of embodiment 29 compound 20b
Prepared through hydroxide palladium carbon catalytic reduction reaction by compound 19b, concrete operation step is with the conjunction of compound 20a Become, yield 88.7%.
1H NMR(600MHz,CDCl3)δ5.02(s,1H,H-1”),4.81(s,1H,H-1’),4.57(s,1H,H-1), 4.31-4.25(m,1H,H-6”a),4.22(br s,1H,H-2’),4.20-4.15(m,1H,H-6”a),4.14-4.10(m, 3H,H-2”,CH2OP), 4.05 (d, J=3.0Hz, 1H, H-2), 3.89-3.84 (m, 3H, H-6a ', OCH 2CH2CH3),3.76- 3.68(m,3H,H-6b’,H-4”,H-4’),3.55-3.42(m,6H),3.26-3.14(m,4H,H-5”,H-5,CH 2NH2), 1.64-1.59(m,2H,OCH2 CH 2CH3), 0.94 (t, J=7.8Hz, 1H, OCH2CH2 CH 3).13C NMR(150MHz,CDCl3)δ 101.4,101.3,100.3,79.4,77.7,76.86,75.4,75.3,73.3,72.8,70.8,70.4,67.7,66.4, 66.1,64.7,64.6,61.9,61.1,60.8,40.0,22.6,9.6.HRMS(ESI):calcd.for C23H44NO19P[M- H]-m/z,668.2167；found,668.2166.
The synthesis of embodiment 30 compound 20c
Prepared through hydroxide palladium carbon catalytic reduction reaction by compound 19c, concrete operation step is with the conjunction of compound 20a Become, yield 93.5%.
1H NMR(600MHz,CDCl3)δ5.06(s,1H,H-1”’),5.01(s,1H,H-1”),4.75(s,1H,H-1’), 4.57 (s, 1H, H-1), 4.49 (d, J=3.0Hz, 1H, H-2 "), 4.39-4.33 (m, 1H, H-6a " '), 4.18-4.06 (m, 5H,H-6b”’,H-2’,H-2”’,CH2OP), 4.05 (d, J=3.0Hz, 1H, H-2 "), 3.89-3.81 (m, 5H), 3.76- 3.39(m,14H),3.37-3.29(m,2H,),3.24-3.11(m,5H,).13C NMR(150MHz,CDCl3)δ102.4, 102.0,100.9,100.4,81.3,80.1,77.1,76.9,75.4,75.4,73.5,72.8,72.7,72.6,70.8, 70.68,67.4,67.3,66.4,64.5,64.49,61.7,61.6,61.1,60.6,40.1,40.0,22.7,9.7.HRMS (ESI):calcd.for C29H54NO24P[M-H]-m/z,830.2695；found,830.2676.
The synthesis of embodiment 31 compound 22a
Compound 20a and Succinimidyl glutarate 21 (DSG, 15equiv) are dissolved in DMF/PBS (4:1,0.1M PBSbuffer) mixed solution, stirs four hours under room temperature, removes solvent and revolve twice with toluene band under low-temp low-pressure, residual Thing adds Excess ethyl acetate repeatedly to wash, and obtains Acibenzolar sterling, yield 78.2% after being dried.
HRMS(ESI):calcd.for C26H42N2O19P[M-H]-m/z,717.2125,found,717.2119.
The synthesis of embodiment 32 compound 22b
Preparation is reacted by compound 20b and Succinimidyl glutarate 21, concrete operation step is with the conjunction of compound 22a Become, yield 81.5%.
HRMS(ESI):calcd.for C32H52N2O24P[M-H]-m/z,879.2653,found,879.2656.
The synthesis of embodiment 33 compound 22c
Preparation is reacted by compound 20c and Succinimidyl glutarate 21, concrete operation step is with the conjunction of compound 22a Become, yield 82.3%.
HRMS(ESI):calcd.for C38H62N2O29P[M-H]-m/z,1041.3187,found,1041.3176.
The synthesis of embodiment 34 glycoprotein conjugate (1a-c, 2a-c)
Compound 22 and carrier protein KLH or HAS are 30 according to mol ratio:1 (Acibenzolar：Carrier protein) it is dissolved in 0.1M PBS cushioning liquid, is stirred at room temperature two and half under argon gas protection.Reactant liquor is through Biogel A 0.5 column chromatography (0.1MPBS delays Rush liquid, pH=7.8), deionized water dialysis and freeze-drying after sterling glycoprotein conjugate 1a-c and 2a-c (molecular weight is shown in attached Fig. 5-7).
Embodiment 35 the compounds of this invention immunocompetence is tested
First, experiment material and source
Test compound：Prepared glycoprotein compound 1a-c in the embodiment of the present invention 34.
2nd, test method
(1) mouse immunization protocol
1) select 6-8 week female Balb/c mouse (being purchased from Chinese Academy of Sciences's Shanghai Experimental Animal Center), according to vaccine+assistant Agent and being only grouped with Vaccination Protocols, control group is only adjuvant group, every group of 6 mouse；
2) by glycoprotein compound 1a-c and its corresponding interpolation adjuvant group (being dissolved in PBS) suspension hypodermic injection mouse, often The glycoprotein of secondary immunity contains the sugar of 2 μ g, uses same method booster immunization again every 2 weeks, altogether immunity 4 times.Before immunity, Latter 13 days of 2nd immunity, latter 13 days of third time immunity and the 4th immunity take blood for latter 14 days, separate serum and are placed in -80 DEG C of ice Case is frozen to be measured.
(2) Serum Antibody titer determination before and after mouse immune
Each every mouse pooled serum titre of experimental group is all detected using ELISA method.
1) wrapper sheet：Compound 2a-c is diluted to 5ug/ml with PH 9.6 carbonate buffer solution respectively and is coated in enzyme mark row's batten On.37 DEG C of combinations 2 hours, use 4 DEG C of confining liquid (15% skim milk) 200ul/ hole overnight afterwards；
2) after PBST washs 3 times, every hole adds 100ul doubling dilution each group mice serum, and 37 DEG C are incubated 2 hours；
3), after PBST washs 5 times, add 100ul 1:Rabbit anti-mouse IgG bis- of the HRP mark after 2000 dilutions resists (Invitrogen), 37 DEG C are incubated 45 minutes；
4), after PBST washs 5 times, after TMB colour developing, ELIASA (MultiscanMK3, Thermo) reads A450 numerical value, is higher than The definition that 3 times of negative control is positive serum.
(3) after mouse immune taken serum and candida albicans cell combination
1) picking candida albicans SC5314 monoclonal bacterium colony YPD fluid nutrient medium increases bacterium 24 hours, now right for growing The yeast like cell of number phase.Candida albicans hyphal phase abductive approach：Logarithmic phase candida albicans will be grown and utilize the resuspended washing 2 of PBS Secondary, it is placed in containing 10% calf serum, 37 DEG C of quiescent culture of 1640 culture medium 90 minutes；
2) by the candida albicans of Hyphal form or Yeast Phase resuspended 2 times with PBS, 10 points are fixed for -20 DEG C with 4% paraformaldehyde Clock, uses resuspended 3 times of PBS again after centrifugation；
3) experimental group and control group serum are pressed 1 with the PBS containing 10% calf serum:100 dilution after respectively with fixation Thalline afterwards is incubated 40 minutes altogether, thereafter with PBS centrifugation, resuspended 3 times；
4) be incubated 30 minutes altogether with sheep anti-mouse igg antibodies and the thalline of 488 marks, thereafter with PBS centrifugation, resuspended 3 times；
5) resuspended bacterium is placed in 96 orifice plates and stands 10 minutes, using fluorescence microscope in 488 observed under fluorescent light；
6) resuspended bacterium is placed in direct flow cytometer detection after fluidic cell centrifuge tube, exciting light selects FITC, profit Returned to zero with negative control candida albicans, again the bacterium after being incubated altogether with fluorescence antibody is carried out upper machine testing, each loading Read 10000 cells.
3rd, experimental result
The KLH- glycoprotein conjugate 1a-c subcutaneous inoculation mouse of present invention preparation, all can induce body to produce stronger exempting from Epidemic disease response, the wherein immunogenicity of compound 1b are the strongest and immune serum that produce being capable of specific recognition candida albicans cell table Face polysaccharide antigen, experimental result is shown in accompanying drawing 8-10.
The above is only the preferred embodiment of the present invention it is noted that ordinary skill people for the art Member, on the premise of without departing from the inventive method, can also make some improvement and supplement, these improve and supplement also should be regarded as Protection scope of the present invention.

Claims (10)

1. a kind of β -1,2-D- oligomerization Mannoproteins conjugate is it is characterised in that described β -1,2-D- oligomerization Mannoproteins Conjugate structure formula is：
Wherein, n is carrier protein KLH or HSA for 0,1 or 2, R.
2. β -1 described in 1 is required according to power, 2-D- oligomerization Mannoproteins conjugate is it is characterised in that form the monose of oligosaccharides Unit is β -1,2-D- mannose.
3. β -1 requiring described in 1 according to power, 2-D- oligomerization Mannoproteins conjugate it is characterised in that by phosphorylation β -1, 2-D- oligomerization mannose is obtained with carrier protein couplet.
4. β -1 requiring described in 3 according to power, 2-D- oligomerization Mannoproteins conjugate it is characterised in that using glutaryl as β -1,2-D- oligomerization the mannose of phosphorylation and the linking group of carrier protein.
5. β -1 described in claim 1, the preparation method of 2-D- oligomerization Mannoproteins conjugate is it is characterised in that include as follows Step：
The first step, synthesizes β -1,2-D- mannobiose acceptor 7, β -1,2-D- manninotriose acceptor 10：
Second step, synthesizes phosphorylation β -1,2-D- oligomerization mannose 19a-c：
3rd step, synthesizes β -1,2-D- oligomerization Mannoproteins conjugate 1a-c, 2a-c：
6. β -1 according to claim 5, the preparation method of 2-D- oligomerization Mannoproteins conjugate is it is characterised in that described Phosphorylation β -1,2-D- oligomerization mannose preparation method is as follows：
(1) with alpha-D-glucose as raw material, β -1,2-D- sweet dew monose acceptor and 3,4,6- are prepared in the presence of reaction promoter Tribenzyl -2- acetoxyl group glucose tri- chloroacetimidate donor；
(2) there is glycosylation preparation two in the monose donor of above-mentioned preparation and monose acceptor under catalyst (TMSOTf) effect Sugar, prepares β -1,2-D- mannobiose acceptor by configuration reversal on 2 '-position of disaccharides；β -1,2-D- mannobiose acceptor and Monose donor prepares β -1,2-D- manninotriose acceptor by preceding method；
(3) β -1,2-D- mannose receptor respectively with 6- tert-butyl diphenyl silicon substrate -3,4- dibenzyl -2- acetoxyl group glucose Tri- chloroacetimidate donor prepares corresponding β -1,2-D- oligomerization mannose through glycosylation；Oligomerization mannose non-reducing end 6- React and phosphorylation β -1,2-D- oligomerization mannose is obtained with phosphorylation agent after TBDPS protection is taken off in position.
7. β -1 according to claim 5, the preparation method of 2-D- oligomerization Mannoproteins conjugate is it is characterised in that include Following steps：
(1) β -1,2-D- oligomerization mannose of phosphorylation and hydroxyl Succinimidyl glutarate are reacted and β -1,2-D- oligomerization is obtained Mannose Acibenzolar；
(2) β -1 prepared by step (1), 2-D- oligomerization mannose Acibenzolar under weak basic condition with carrier protein couplet, obtain To β -1,2-D- oligomerization Mannoproteins conjugate.
8. the arbitrary described β -1,2-D- oligomerization Mannoproteins conjugate of claim 1-4 is true in preparation prevention and/or treatment Application in the medicine of bacterium infection.
9. application according to claim 8 is it is characterised in that described fungi is candida albicans.
10. a kind of antifungal fungi vaccine, including arbitrary described β -1 of claim 1-4 of therapeutically effective amount, 2-D- is few Poly- Mannoproteins conjugate and pharmaceutically acceptable auxiliary material or adjuvant.
CN201610928843.7A 2016-10-31 2016-10-31 Beta-1,2-D-oligomeric mannoprotein conjugates and preparation method and application thereof Pending CN106432371A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610928843.7A CN106432371A (en) 2016-10-31 2016-10-31 Beta-1,2-D-oligomeric mannoprotein conjugates and preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610928843.7A CN106432371A (en) 2016-10-31 2016-10-31 Beta-1,2-D-oligomeric mannoprotein conjugates and preparation method and application thereof

Publications (1)

Publication Number Publication Date
CN106432371A true CN106432371A (en) 2017-02-22

Family

ID=58178082

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610928843.7A Pending CN106432371A (en) 2016-10-31 2016-10-31 Beta-1,2-D-oligomeric mannoprotein conjugates and preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN106432371A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110724189A (en) * 2019-11-04 2020-01-24 济南山目生物医药科技有限公司 (1-5) -Gal- (1-6) -Man glycoprotein and preparation method and application thereof
JP2021528371A (en) * 2018-05-09 2021-10-21 ザ・ユニバーシティ・オブ・シカゴＴｈｅ Ｕｎｉｖｅｒｓｉｔｙ Ｏｆ Ｃｈｉｃａｇｏ Compositions and Methods for Immune Tolerance

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1401786A (en) * 2001-11-15 2003-03-12 青岛海洋大学 Process for preparing beta-D-mannuronic acid oligosaccharide monoclonal antibody
CN1743009A (en) * 2005-09-20 2006-03-08 中国人民解放军第二军医大学 Medicine carrier pseudoglucoprotein nano particle of double-target anti-liver-fibrosis and its preparing method
CN102382154A (en) * 2011-11-09 2012-03-21 中国人民解放军第二军医大学 Saponin compound, preparation method for same, and application thereof in immunologic adjuvant preparation
CN103432566A (en) * 2008-12-16 2013-12-11 建新公司 Oligosaccharide-protein conjugates

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1401786A (en) * 2001-11-15 2003-03-12 青岛海洋大学 Process for preparing beta-D-mannuronic acid oligosaccharide monoclonal antibody
CN1743009A (en) * 2005-09-20 2006-03-08 中国人民解放军第二军医大学 Medicine carrier pseudoglucoprotein nano particle of double-target anti-liver-fibrosis and its preparing method
CN103432566A (en) * 2008-12-16 2013-12-11 建新公司 Oligosaccharide-protein conjugates
CN102382154A (en) * 2011-11-09 2012-03-21 中国人民解放军第二军医大学 Saponin compound, preparation method for same, and application thereof in immunologic adjuvant preparation

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HONG XIN，等: "Synthetic glycopeptide vaccines combining β-mannan and peptide epitopes induce protection against candidiasis", 《PNAS》 *
廖俊: "β-[1,2]-寡聚甘露糖肽化合物的合成及白念珠菌疫苗的免疫活性研究", 《第二军医大学博士学位论文》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2021528371A (en) * 2018-05-09 2021-10-21 ザ・ユニバーシティ・オブ・シカゴＴｈｅ Ｕｎｉｖｅｒｓｉｔｙ Ｏｆ Ｃｈｉｃａｇｏ Compositions and Methods for Immune Tolerance
CN110724189A (en) * 2019-11-04 2020-01-24 济南山目生物医药科技有限公司 (1-5) -Gal- (1-6) -Man glycoprotein and preparation method and application thereof
WO2021087709A1 (en) * 2019-11-04 2021-05-14 济南山目生物医药科技有限公司 (1-5)-gal-(1-6)-man glycoprotein, preparation method therefor and use thereof
CN110724189B (en) * 2019-11-04 2022-01-25 济南山目生物医药科技有限公司 (1-5) -Gal- (1-6) -Man glycoprotein and preparation method and application thereof

Similar Documents

Publication Publication Date Title
CA2606890C (en) Immunogens for meningitidis-a vaccines
EP0276516B1 (en) New oligosaccharides, immunogens and vaccines, and methods for preparing such oligosaccharides, immunogens and vaccines
JP7208995B2 (en) Method for preparing the outer core octacarbon sugar of Helicobacter pylorilipopolysaccharide
KR20160055154A (en) Protein and peptide-free synthetic vaccines against steptococcus pneumoniae type 3
CA3140411C (en) Saponin conjugate and vaccine or pharmaceutical composition comprising the same
CA2961694A1 (en) Vaccines against streptococcus pneumoniae serotype 8
Sanders et al. Synthesis of sulfated trisaccharide ligands for the selectins
CN106432371A (en) Beta-1,2-D-oligomeric mannoprotein conjugates and preparation method and application thereof
EP3274358B1 (en) Vaccine against carbapenem-resistantklebsiella pneumoniae
DK2164521T3 (en) GLYCOCONJUGATES AND USE THEREOF AS VACCINE AGAINST SHIGELLA FLEXNERI SEROTYPE 3A AND X
JP6622804B2 (en) Vaccine against Streptococcus pneumoniae serotype 4
CN106366163A (en) Beta-1,2-D-oligomeric mannose peptide-protein conjugate and preparation method and application thereof
CN100365007C (en) New type trisaccharide and penta saccharid oligo saccharide antigen, their synthesis method and application in preparation of medicine for inhibiting exclusion reaction
WO2022106703A1 (en) Protected disaccharides, their process of preparation and their use in the synthesis of zwitterionic oligosaccharides, and conjugates thereof
US20220152183A1 (en) Carbocyclic derivatives and conjugated derivatives thereof, and their use in vaccines
CN106084037B (en) Bacillus anthracis capsule surface trisaccharide conjugate and preparation method and application thereof
AU2003248927A1 (en) Solid-phase and solution-phase synthesis of glycosylphosphatidylinositol glycans
CN113166186A (en) Stable vaccine against clostridium difficile
WO2014067970A1 (en) Glycoconjugates and their use as potential vaccines against infection by shigella flexneri
US20230346905A1 (en) Pentavalent vaccine against neisseria meningitidis comprising a synthetic men a antigen
Auberger Synthesis of Neisseria meningitidis serogroup A carba analogues as hydrolytically stable antigens for antimeningococcal glycoconjugate vaccines
JPH0680702A (en) Acidic sugar derivative
Tang Automated solution-phase synthesis of β-mannans
WO2020135898A1 (en) Synthetic oligosaccharide derivatives as a vaccine against bordetella pertussis
CN1334268A (en) Process for synthesizing active tetrose fragment of arabogalactan of medical purpose

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20170222

WD01 Invention patent application deemed withdrawn after publication