CN102617662B - Amino pentosaccharide related with nitrogen fixation activity of plants, and preparation method and application thereof - Google Patents
Amino pentosaccharide related with nitrogen fixation activity of plants, and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses an amino pentosaccharide compound related with nitrogen fixation activity of plants, and a preparation method and application thereof. The structural formula of the compound is shown in formula I. The preparation method comprises respectively enabling acyl glucose and acyl mannose to serve as a glycosyl donor, enabling rhamnose of 1-O-P-methoxyphenyl to serve as a glycosyl receptor, first obtaining disaccharide of 1-3-beta-connetion and disaccharide of 1-2-alpha-connection, enabling the disaccharide of 1-2-alpha-connection to serve as a glycosyl receptor and enabling acyl rhamnose to serve as a glycosyl donor so as to obtain trisaccharide, enabling the trisaccharide to serve as a glycosyl donor and enabling the disaccharide of 1-3-beta-connetion to serve as a glycosyl receptor so as to obtain pentosaccharide, and obtaining the target compound I by removing protection. Indoor biological activity experiments and filed test results prove that the compound I has the effects of adjusting growth, promoting growth of a root system, increasing the total root number of a single plant, promoting seedling growth, adding single-plant tillering and cultivating strong seedlings.
Description
Technical field
The present invention relates to a kind of amino pentasaccharides relevant with plant nitrogenase activity and preparation method thereof and application.
Background technology
Current multiple oligosaccharide kind material has been successfully applied in agriculture production, and for example chitin can be used as foliage fertilizer, can promote root growth, strain to strengthen full of leaves, and the phase of yielding positive results is played short flower, swollen fruit effect.Oligochitosan not only can stimulating plant growth, make farm crop and fruit and vegetable volume increase good harvest, disease resistance that also can inducing plant, produces immunity and killing action to multiple fungi, bacterium and virus.Agricultural chemicals scientific workers are more and more to the concern of oligosaccharide kind agricultural chemicals at present, and the oligosaccharides of some synthetic, as glucohexaose, has also been proved to be the activation capability with plant ego defense system.
The growth needs of plant constantly absorbs ammonium salt and nitrate from soil around, so by these inorganic nitrogens with changing into the organonitrogens such as protein in plant materials.Root nodule bacterium fixed nitrogen is the important sources that many plants obtain nitrogen element, and root nodule bacterium and host plant identify and form symbiotic relationship mutually by the transmission of signaling molecule, and carbohydrate is one (Battisti, L. the most key in these signaling molecules; Lara, J.C.; Leigh, J.A.Proc.Natl.Acad.Sci.U.S.A.1992,89,5625-5629.).There are some researches show, part root nodule bacterium oligosaccharides has the function that stimulates plant root nodule to form.In some cases, even without the existence of root nodule bacterium, as long as inject lower concentration (10 in host plant root
-6-10
-12m) root nodule bacterium oligosaccharides, just can cause plant root nodule process.Thereby just there are very important Research Significance and potential using value synthesizing of these oligosaccharides.
(Wang, the Y. such as Ying Wang in 1994, Hollingsworth, R.I.The structure of the O-antigenicchain of the lipopolysaccharide of Rhizobium trifokii 4s.Carbohydrate.Research, 1994, 260:305-317.) determined in vinelandii Rhizobium trifokii 4s with the closely-related lipopolysaccharides of nitrogen fixation in the structure of O-specificity chain, its structure be take 2-acetylaminohydroxyphenylarsonic acid 2-deoxidation-α-D-MANNOSE-(1 → 2)-[α-L-rhamnosyl-(1 → 3)-] α-L-rhamnosyl-(1 → 4)-2-acetylaminohydroxyphenylarsonic acid 2-deoxidation-β-D-Glucose-(1 → 3)-α-L-rhamnosyl is the polysaccharide polymer polymkeric substance of repeating unit, under structural formula is shown in:
The repeating unit oligosaccharides of polysaccharide is the bioactive basic structure of performance, has not yet to see the synthetic method of pentasaccharides unit and the report of application of this polysaccharide.
Summary of the invention
The object of this invention is to provide a kind of amino pentasaccharides compound and preparation method thereof.
Amino five compounds provided by the present invention, its chemical name is: 2-acetylaminohydroxyphenylarsonic acid 2-deoxidation-α-D-MANNOSE-(1 → 2)-[α-L-rhamnosyl-(1 → 3)-] α-L-rhamnosyl-(1 → 4)-2-acetylaminohydroxyphenylarsonic acid 2-deoxidation-β-D-Glucose-(1 → 3)-α-L-rhamnosyl p-methoxyphenyl glycosides, and structural formula is suc as formula shown in I:
(formula I)
Preparation method comprises the steps:
1) three sugar compounds donors shown in the disaccharide compound acceptor shown in formula II and formula III are carried out to condensation reaction, obtain the amino pentasaccharides compound of the full guard shown in formula IV;
Wherein, Ac represents that ethanoyl, Bz represent benzoyl;
2) make the amino pentasaccharides compound of the full guard shown in formula IV carry out protecting group in saturated methanol ammonia solution and remove reaction, obtain compound shown in formula I.
Wherein, step 1) the disaccharide compound acceptor shown in Chinese style II and the mol ratio of three sugar compounds donors shown in formula III can be 1: 1-1.5.
Step 1) described condensation reaction carries out under the katalysis of TMS triflate (TMSOTf), and the mol ratio of three sugar compounds donors shown in described TMS triflate and formula III is 0.01-0.5: 1; The reaction medium of described condensation reaction is methylene dichloride; The temperature of reaction of described condensation reaction can be-20 ℃-0 ℃, and the reaction times can be 2-4 hour.
Step 2) described in reaction temperature of reaction be 20 ℃-30 ℃, the reaction times is 4-8 days.
Disaccharide compound acceptor shown in Chinese style II of the present invention can prepare by the following method:
Take glucose tribromo-acetyl imines ester (1) as glycosyl donor; take acyl group rhamnoside (2) as glycosyl acceptor; carry out condensation reaction and obtain 1 → 3-β-connection disaccharide (3); this disaccharide is through deacetylation, 4; 6-hydroxyl isopropylidene, hydrazinolysis, acetylize, the protection of solution fork base, then selectivity 6-position benzoylation obtains disaccharide (II).
Three sugar compounds donors in the present invention shown in formula III can prepare by the following method:
Take seminose tribromo-acetyl imines ester (9) as glycosyl donor, acyl group rhamnoside (10) is glycosyl acceptor, carries out condensation reaction and obtains 1 → 2-α-connection disaccharide 11, and the de-allyloxycarbonyl of this disaccharide obtains disaccharide 12; 12 obtain with acyl group rhamnosyl tribromo-acetyl imines ester (13) condensation the trisaccharide (14) that (1 → 2)-[α-(1 → 3)-] α-L-is connected; this trisaccharide is through nitrine reduction, acetylize, de-p-methoxyphenyl, and then activation is three saccharide donors (III).
Concrete, the synthetic method of 2-acetylaminohydroxyphenylarsonic acid 2-of the present invention deoxidation-α-D-MANNOSE-(1 → 2)-[α-L-rhamnosyl-(1 → 3)-] α-L-rhamnosyl-(1 → 4)-2-acetylaminohydroxyphenylarsonic acid 2-deoxidation-β-D-Glucose-(1 → 3)-alpha-L-rhamnoside can be carried out by the following method:
Synthesizing of disaccharide compound acceptor shown in formula II:
Its reaction process as shown in Figure 1.
Take the glucose tribromo-acetyl imines ester (1) of 1～1.5 mole as glycosyl donor, the rhamnosyl (2) of the 1-O-p-methoxyphenyl protection of 1 mole is glycosyl acceptor, after being mixed, both with methylene dichloride, dissolve, under the catalysis of TMS triflate (TMSOTf), in-20 ℃～0 ℃, carry out condensation reaction, react after approximately 2～4 hours, the disaccharide (3) of preparing 1 → 3-β-connection, the mol ratio of TMSOTf and glycosyl donor (1) is 0.01～0.5: 1; Be dissolved in Acetyl Chloride 98Min./methyl alcohol (1%, v/v) solution 3, react 10～12 hours, selectivity deacetylate, obtains trihydroxy-disaccharide 4; Be dissolved in dimethyl formamide 4, at a hydration p-methyl benzenesulfonic acid (p-TsOHH
2o) under catalysis, at room temperature react after 2～4 hours with 2-methoxyl group propylene, obtain the disaccharide 5 of 4,6-position selectivity isopropylene protection, p-TsOHH
2the mol ratio of O and disaccharide 4 is 0.015～0.02: the mol ratio of 1,2-methoxyl group propylene and disaccharide 4 is 1～1.5: 1; Be dissolved in ethanol/85% hydrazine hydrate (9: 1, v/v) 5, reflux 24～48 hours, obtains the disaccharide 6 of deprotection; Be dissolved in anhydrous pyridine 6, add after diacetyl oxide, at 0 ℃～30 ℃, react 12～24 hours, obtain the disaccharide 7 of ethanoyl protection, the mol ratio of diacetyl oxide and compound 6 is 7～9: 1; Be dissolved in acetic acid/water (70%, v/v) 7, in 70～80 ℃ of reactions 1～2 hour, obtain the disaccharide 8 of de-fork base protection; Be dissolved in anhydrous pyridine 8, add after Benzoyl chloride, in-20 ℃～0 ℃ reaction 2～5 hours, obtain disaccharide II, the mol ratio of Benzoyl chloride and compound 8 is 1～1.1: 1.
Synthesizing of three sugar compounds donors shown in formula III:
Its reaction process as shown in Figure 2.
Take the seminose tribromo-acetyl imines ester 9 of 1～1.5 mole as glycosyl donor, the rhamnosyl 10 of the 1-O-p-methoxyphenyl protection of 1 mole is glycosyl acceptor, after being mixed, both with methylene dichloride, dissolve, under the catalysis of TMS triflate (TMSOTf), in-20 ℃～0 ℃, carry out condensation reaction, react after approximately 2～4 hours, prepare the disaccharide 11 of 1 →-α-connection, the mol ratio of TMSOTf and glycosyl donor 9 is 0.01～0.5: 1; By 11, be dissolved in methyl alcohol/tetrahydrofuran (THF) (50%; v/v) in solution; add ammonium acetate, four triphenyl phosphorus palladium and sodium borohydrides; in-20 ℃～0 ℃, react 2～10 minutes; obtain the disaccharide 12 of de-allyloxycarbonyl protection; the mol ratio of ammonium acetate and compound 11 is that the mol ratio of 10～20: 1, four triphenyl phosphorus palladium and compound 11 is 0.04～0.05: 1, and the mol ratio of sodium borohydride and compound 11 is 1.1～2: 1.
Take the rhamnosyl tribromo-acetyl imines ester 13 of 1～1.5 mole as glycosyl donor, the compound 12 of 1 mole is glycosyl acceptor, after being mixed, both with methylene dichloride, dissolve, under the catalysis of TMS triflate (TMSOTf), in-20 ℃～0 ℃, carry out condensation reaction, react after approximately 2～4 hours, the trisaccharide (14) that prepare (1 → 2)-[α-(1 → 3)-] α-L-connects, the mol ratio of TMSOTf and glycosyl donor 13 is 0.01～0.5: 1; By 14, be dissolved in methyl alcohol, add palladium/carbon catalyst, pass into a normal atmosphere hydrogen, in 0 ℃～30 ℃, react 4～8 hours, gained compound is dissolved in anhydrous pyridine, adds after diacetyl oxide, at 0 ℃～30 ℃, react 12～24 hours, the trisaccharide 15 that obtains ethanoyl protection, the mol ratio of diacetyl oxide and midbody compound is 1.5～2: 1; By 15, be dissolved in acetonitrile/water (50%, v/v) in solution, add ceric ammonium nitrate, in 10 ℃～30 ℃, react 20～50 minutes, obtain the product of de-p-methoxyphenyl, this intermediate is dissolved in methylene dichloride, add Trichloroacetonitrile, at DBU (1,8-diazabicylo [5.4.0] 11 carbon-7-alkene) under catalysis, 10 ℃～30 ℃ are reacted 1～3 hour, obtain three saccharide donor III, and the mol ratio of Trichloroacetonitrile and DBU and midbody compound is respectively 1.5～2: 1 and 0.2～0.3: 1.
Synthesizing of target pentasaccharides shown in formula I:
Its reaction process as shown in Figure 3.
After being mixed, the disaccharide II of the three saccharide donor III of 1～1.5 mole and 1 mole dissolves with methylene dichloride, under the catalysis of TMS triflate (TMSOTf), in-20 ℃～0 ℃, carry out condensation reaction, react after approximately 2～4 hours, prepare full guard pentasaccharides IV, the mol ratio of TMSOTf and glycosyl donor III is 0.01～0.5: 1; IV is dissolved in saturated methyl alcohol/ammonia solution, reacts 4～8 days in 20 ℃～30 ℃, obtain the amino pentasaccharides I of target of deprotection.
A further object of the present invention is to provide the application of compound shown in formula I.
The application of compound shown in formula I provided by the present invention is that it is in the application of preparing in plant-growth regulator.
Described plant-growth regulator has following at least one effect: 1) promote root growth, 2) promote growth of seedling, 3) increase individual plant total root number, 4) increase individual plant and tiller, 5) cultivate strong sprout.
Described plant-growth regulator specifically can be used for regulating the growth regulating of the plants such as wheat, cotton.
In addition, the plant-growth regulator take the amino pentasaccharides shown in formula I as active fraction preparation also belongs to protection scope of the present invention.
The present invention is from rhamnosyl and glucosamine hydrochloride; take the synthesis strategy of 2+3; first synthetic disaccharide and three saccharide donors, then adopt the condensation of Schmidt glycosidation to obtain pentasaccharides, and this pentasaccharides obtains the amino pentasaccharides of target compound after deprotection.The suitable blocking group of ingenious introducing in reaction designing, efficiently carries out whole reaction process, and easy and simple to handle.Indoor biological activity test and field test results show, amino pentasaccharides provided by the invention has growth regulation to wheat, cotton under finite concentration, promotes the growth of root system, increase the total root number of individual plant, promote growth of seedling, add individual plant and tiller, cultivate the effect in strong sprout.
Accompanying drawing explanation
Fig. 1 is the synthetic schemes of the disaccharide compound acceptor shown in formula II.
Fig. 2 is the synthetic schemes of three sugar compounds donors shown in formula III.
Fig. 3 is the synthetic schemes of the target pentasaccharides shown in formula I.
Fig. 4 is the hydrogen nuclear magnetic resonance spectrogram of compound shown in formula IV.
Fig. 5 is the carbon-13 nmr spectra figure of compound shown in formula IV.
Fig. 6 is the hydrogen nuclear magnetic resonance spectrogram of compound shown in formula I.
Fig. 7 is the carbon-13 nmr spectra figure of compound shown in formula I.
Embodiment
Below by specific embodiment, the present invention will be described, but the present invention is not limited thereto.
Experimental technique described in following embodiment, if no special instructions, is ordinary method; Described reagent and material, if no special instructions, all can obtain from commercial channels.
Initial action starting compound the 1,2,9,10, the 13rd used in embodiment, prepares according to the method for following document respectively:
Compound 9 synthesized reference document: Briner, K.; Vasella, A.Glycosylphosphonates of2-Amino-2-deoxy-aldoses.Synthesis of a Phosphonate Analogue of Lipid X
1.Helv.Chim.Acta 1987,70:1341-1356.
Synthesizing of embodiment 1, disaccharide 3
By compound 1 (3.2g, 5.5mmol), compound 2 (2.4g, 5.0mmol) and
molecular sieve (3g) is dissolved in 100mL methylene dichloride; ice bath is cooled to-10 ℃; under nitrogen protection, stir 0.5 hour; add TMS triflate 54 μ L (0.3mmol); keep-10 ℃ of reactions 0.5 hour; remove ice bath and make reactant naturally be warmed up to room temperature, after 2 hours, add 100 μ L triethylamines and finish reaction.Suction filtration reaction solution, concentrated under filtrate decompression, column chromatography for separation (sherwood oil: ethyl acetate=2: 1, v/v) obtains white foam shape solid disaccharide 3 (3.4g, 76%).[α]
D 25+9.6°(c?0.5?CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.13-7.16 (m, 14H, Bz-H, Phth-H), 7.06-7.03 (m, 2H, Ar-H), 5.67 (m, 1H, H-2), 5.65 (d, 1H, J8.4Hz, H-1 '), 5.64 (t, 1H, J
2 ', 3 ', J
3 ', 4 '10.6Hz, H-3 '), 5.56 (d, 1H, J 1.6Hz, H-1), 5.43 (t, 1H, J9.8Hz, H-4), 5.03 (t, 1H, J9.9Hz, H-4 '), 5.49 (dd, 1H, J
2,33.7Hz, J
3,49.7Hz, H-3), 4.26 (dd, 1H, J
1 ', 2 '8.4Hz, J
2 ', 3 '10.7Hz, H-2 '), 4.20-4.11 (m, 2H, 2 × H-6 '), 4.04,3.89 (2m, 2H, 2 × H-5), 3.78 (s, 1H, C
6h
4oCH
3), 1.96,1.88,1.72 (3s, 9H, 3 × CH
3cO), 1.09 (d, 1H, J6.2Hz, H-6).
Synthesizing of embodiment 2, disaccharide 4
By disaccharide 3 (1.8g, 2.0mmol) be dissolved in containing in the methanol solution (40mL) of 1% Acetyl Chloride 98Min., after 12 hours, react completely, neutralize with triethylamine, reaction solution decompression is lower concentrated, column chromatography for separation (sherwood oil: ethyl acetate=1: 2) obtain white solid disaccharide 4 (1.4g, 90%).[α]
D 25-17.0°(c?0.9CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.11-7.10 (m, 14H, Bz-H, Phth-H), 7.06-7.03 (m, 2H, Ar-H), 6.85-6.82 (m, 2H, Ar-H), 5.87 (dd, 1H, J
1,21.8Hz, J
2,33.5Hz, H-2), 5.46 (d, 1H, J 8.2Hz, H-1 '), 5.42 (d, 1H, J 1.6Hz, H-1), 5.38 (t, 1H, J 9.7Hz, H-4), 4.42 (dd, 1H, J
2,33.7Hz, J
3,49.7Hz, H-3), 4.11-4.01 (m, 2H, 2 × H-5), 3.92-3.82 (m, 2H, H-3 ', OH), 3.77 (s, 1H, C
6h
4oCH
3), 3.61 (m, 1H, H-2 '), 3.46 (m, 1H, H-4 '), 3.38-3.30 (m, 2H, 2 × H-6 '), 3.24 (t, 1H, J 7.5Hz, OH), 2.98 (d, 1H, J 5.8Hz, OH), 1.09 (d, 1H, J 6.2Hz, H-6).
Synthesizing of embodiment 3, disaccharide 5
Disaccharide 4 (1.3g, 1.7mmol) is dissolved in 20mL dry DMF, under stirring, adds a hydration tosic acid (6.4mg, 0.034mmol) and 2-methoxyl group propylene (0.31mL, 2.5mmol).After 3 hours, react completely, by lower reaction solution decompression concentrated, revolve steaming with toluene (2 × 10mL) azeotropic.Enriched material column chromatography for separation (sherwood oil: ethyl acetate=2: 1) obtain white foam shape solid disaccharide 5 (1.2g, 88%).[α]
D 25-25.0°(c?0.6?CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.14-7.21 (m, 14H, Bz-H, Phth-H), 7.06-7.03 (m, 2H, Ar-H), 6.85-6.82 (m, 2H, Ar-H), 5.59 (dd, 1H, J
1,21.9Hz, J
2,33.6Hz, H-2-Rhap), 5.53 (d, 1H, J 1.8Hz, H-1-Rhap), 5.45 (d, 1H, J 8.2Hz, H-1-Glup), 5.40 (t, 1H, J 9.8Hz, H-4-Rhap), 4.48 (dd, 1H, J
2,33.6Hz, J
3,49.7Hz, H-3-Rhap), 4.29 (m, 1H, H-2-Glup), 4.12-3.91 (m, 3H, H-3-Glup, H-4-Glup, H-5), 3.77 (s, 1H, C
6h
4oCH
3), 3.59 (m, 1H, H-5), 3.47-3.42 (m, 2H, 2 × H-6-Glup), 2.14 (d, 1H, J 3.6Hz, OH), 1.40,1.37 (2s, 6H, Me
2c), 1.08 (d, 1H, J6.3Hz, H-6).
Synthesizing of embodiment 4, disaccharide 7
By disaccharide 5 (1.1g, 1.4mmol) be dissolved in 4mL 85% hydrazine hydrate and 36mL ethanol, stirring and refluxing reaction at 90 ℃, after 48 hours, react completely, by cooling reaction solution rear concentrating under reduced pressure, with toluene (2 × 10mL) azeotropic, revolve to steam and obtain oily enriched material 6, this enriched material is dissolved in 10mL pyridine, add 3mL diacetyl oxide, under room temperature, react, after 12 hours, react completely, by reaction solution concentrating under reduced pressure, revolve steaming with toluene (2 × 10mL) azeotropic, enriched material column chromatography for separation (sherwood oil: ethyl acetate=1: 1, v/v) obtain white foam shape solid disaccharide 7 (0.69g, 79%).[α]
D 25-3.8°(c?0.3?CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 6.98-6.93 (m, 2H, Ar-H), 6.84-6.80 (m, 2H, Ar-H), 5.72 (m, 1H, NHAc), 5.32-5.30 (m, 2H, H-1-Rhap, H-2-Rhap), 5.14 (t, 1H, J
3,4, J
4,59.6Hz, H-4-Rhap), 5.10 (t, 1H, J
2 ', 3 ', J
3 ', 4 '9.8Hz, H-3-Glup), 4.71 (d, 1H, J 8.2Hz, H-1-Glup), 4.22 (dd, 1H, J
2,32.8Hz, J
3,49.6Hz, H-3-Rhap), 3.37-3.77 (m, 7H, H-2-Glup, 2 × H-6-Glup, H-5, C
6h
4oCH
3), 3.71 (t, 1H, J
3 ', 4 ', J
4 ', 5 '9.6Hz, H-4-Glup), 2.16,2.10,2.07,1.95 (4s, 12H, 4 × CH
3cO), 1.47,1.38 (2s, 6H, Me
2c).
Synthesizing of embodiment 5, disaccharide 8
By disaccharide 7 (0.62g, 0.97mmol) be dissolved in containing in 70% acetic acid aqueous solution (20mL), stirring reaction at 75 ℃, after 1.5 hours, react completely, concentrated under the cooling rear decompression of reaction solution, revolve steaming, enriched material column chromatography for separation (sherwood oil: ethyl acetate=1: 3 with toluene (2 × 10mL) azeotropic, v/v) obtain white solid disaccharide 8 (0.50g, 86%).[α]
D 25-40.2°(c?0.4?CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 6.98 (d, 2H, J9.1 Hz, Ar-H), 6.82 (d, 2H, J 9.1Hz, Ar-H), 6.12 (d, 1H, J 8.6Hz, NHAc), 5.55 (dd, 1H, J
1,21.8Hz, J
2,33.4Hz, H-2-Rhap), 5.27 (d, 1H, J 1.5Hz, H-1-Rhap), 5.20 (t, 1H, J
2 ', 3 ', J
3 ', 4 '9.3Hz, H-3-Glup), 5.08 (t, 1H, J
3,4, J
4,59.7Hz, H-4-Rhap), 4.91 (d, 1H, J 8.2Hz, H-1-Glup), 4.19 (dd, 1H, J
2,33.5Hz, J
3,49.7Hz, H-3-Rhap), 3.94-3.89 (m, 3H, 2 × H-6-Glup, H-2-Glup), 3.77 (s, 3H, C
6h
4oCH
3), 3.66-3.63 (m, 3H, 2 × OH, H-4-Glup), 3.51 (m, 1H, H-5), 3.33 (m, 1H, H-5), 2.18,2.10,2.08,1.94 (4s, 12H, 4 × CH
3cO), 1.16 (d, 3H, J 6.2Hz, H-6-Rhap).
13c NMR (75MHz, CDCl
3): δ 171.9,171.3,170.4,169.8 (4 × MeCO), 155.2,149.7,117.7 (2), 114.6 (2) (C
6h
4oCH
3), 101.1,96.4 (2 × C-1), 75.8,75.1,72.0,71.2,68.6,68.5,66.8,55.6 (C
6h
4oCH
3), 55.0,23.2,21.1,20.9 (2) (4 × CH
3cO), 17.4 (C-6-Rhap).
By disaccharide 8 (0.38g, 0.63mmol) be dissolved in 5mL pyridine, ice bath is cooled to-10 ℃, in 30 minutes, drip Benzoyl chloride (0.081mL, pyridine solution (3mL) 0.70mmol), remove ice bath and make reactant naturally be warmed up to room temperature, after 2 hours, react completely, add 1mL methyl alcohol termination reaction.By lower reaction solution decompression concentrated, enriched material column chromatography for separation (sherwood oil: ethyl acetate=1: 2, v/v) obtains white foam shape solid disaccharide II (0.36g, 81%).[α]
D 25-52.1°(c?0.8CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.08-8.05 (m, 2H, Bz-H), 7.51 (m, 1H, Bz-H), 7.38-7.33 (m, 2H, Bz-H), 6.89 (d, 2H, J9.1Hz, Ar-H), 6.78 (d, 2H, J 9.1Hz, Ar-H), 5.87 (d, 1H, J 8.5Hz, NHAc), 5.46 (dd, 1H, J
1,22.0Hz, J
2,33.4Hz, H-2-Rhap), 5.33 (d, 1H, J 1.8Hz, H-1-Rhap), 5.28 (t, 1H, J
2 ', 3 ', J
3 ', 4 '9.3Hz, H-3-Glup), 5.11 (t, 1H, J
3,4, J
4,59.7Hz, H-4-Rhap), 4.92 (d, 1H, J 8.2Hz, H-1-Glup), 4.67 (dd, 1H, J 4.5Hz, 12.2Hz, H-6-Glup), 4.56 (dd, 1H, J 1.9Hz, 12.2Hz, H-6-Glup), 4.27 (dd, 1H, J
2,33.5Hz, J
3,49.7Hz, H-3-Rhap), 3.90 (m, 1H, H-2-Glup), 3.77 (s, 3H, C
6h
4oCH
3), 3.70-3.54 (m, 4H, 2 × H-5, OH, H-4-Glup), 2.12,2.10,2.08,1.95 (4s, 12H, 4 × CH
3cO), 1.16 (d, 3H, J 6.2Hz, H-6-Rhap).
13c NMR (75MHz, CDCl
3): δ 171.7,170.4,170.2,169.8 (4 × CH
3cO), 167.0 (C
6h
5cO), 155.1,149.8,117.7 (2), 114.6 (2) (C
6h
4oCH
3), 100.8,96.3 (2 × C-1), 75.0,74.6,74.0,72.2,71.4,68.9,66.7,63.6,55.5 (C
6h
4oCH
3), 55.1,23.1,20.9 (2), 20.8, (4 × CH
3cO), 17.3 (C-6-Rhap).
Synthesizing of embodiment 7, disaccharide 11
By compound 9 (4.1g, 8.6mmol), compound 10 (3.6g, 7.9mmol) and
molecular sieve (4g) is dissolved in 100mL methylene dichloride; ice bath is cooled to-10 ℃; under nitrogen protection, stir 0.5 hour; add TMS triflate 54 μ L (0.3mmol); keep-10 ℃ of reactions 0.5 hour; remove ice bath and make reactant naturally be warmed up to room temperature, after 2 hours, add 100 μ L triethylamines and finish reaction.Suction filtration reaction solution, concentrated under filtrate decompression, column chromatography for separation (sherwood oil: toluene: ethyl acetate=4: 6: 1, v/v/v) must white solid disaccharide 11 (5.2g, 86%).[α]
D 25+55.9°(c?0.3?CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.04-8.02 (m, 2H, Bz-H), 7.57 (m, 1H, Bz-H), 7.47-7.42 (m, 2H, Bz-H), 7.06-7.03 (m, 2H, Ar-H), 6.87-6.84 (m, 2H, Ar-H), 5.69 (m, 1H, CH
2=CHCH
2oCO), 5.53 (dd, 1H, J
2 ', 3 '3.4Hz, J
3 ', 4 '10.1Hz, H-3 '), 5.46-5.37 (m, 4H, H-1 ', H-3, H-4, H-4 '), 5.23-5.06 (m, 2H, CH
2=CHCH
2oCO), 4.96 (d, 1H, J
1,21.5Hz, H-1), 4.52-4.49 (m, 2H, CH
2=CHCH
2oCO), 4.36 (dd, 1H, J
1 ', 2 '2.0Hz, J
2 ', 3 '3.3Hz, H-2 '), 4.32-4.23 (m, 2H, H-5, H-5 '), 4.22 (dd, 1H, J
1,21.7Hz, J
2,33.2Hz, H-2), 4.17-4.02 (m, 2H, 2 × H-6 '), 2.12,2.09,2.07 (3s, 9H, 3 × CH
3cO), 1.28 (d, 1H, J6.3 Hz, H-6).
13cNMR (75MHz, CDCl
3): δ 170.7,169.7,169.6 (3 × COCH
3), 165.4 (COPh), 165.4,154.2,130.8,119.2 (CH
2=CHCH
2oCO), 97.6,96.2 (2 × C-1), 55.6 (C
6h
4oCH
3), 20.6 (2), 20.4 (3 × COCH
3), 17.5 (C-6).
Synthesizing of embodiment 8, disaccharide 12
By disaccharide 11 (4.0g, 5.2mmol) be dissolved in 100mL methyl alcohol/tetrahydrofuran (THF) (v/v=1/1), be cooled to-5 ℃, add ammonium acetate (4.0g, 52mmol), under vigorous stirring, add successively sodium borohydride (47mg, 1.3mmol), four triphenyl phosphorus palladiums (240mg, 0.21mmol) and sodium borohydride (234mg, 6.5mmol), after 1 minute, react completely.By lower reaction solution decompression concentrated, methylene dichloride (150mL) dilution, water (50mL) are washed, organic phase anhydrous Na
2sO
4dry, evaporating column chromatographic separation (sherwood oil: ethyl acetate=3: 1, v/v) obtains white foam shape solid disaccharide 12 (3.3g, 92%).[α]
D 25+2.4°(c?0.6CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.07-8.05 (m, 2H, Bz-H), 7.58 (m, 1H, Bz-H), 7.48-7.43 (m, 2H, Bz-H), 7.05-7.01 (m, 2H, Ar-H), 6.87-6.83 (m, 2H, Ar-H), 5.48 (dd, 1H, J
2 ', 3 '3.8Hz, J
3 ', 4 '9.4Hz, H-3 '), 5.45 (d, 1H, J
1,21.4Hz, H-1), 5.34 (t, 1H, J
3 ', 4 ', J
4 ', 5 '9.4Hz, H-4 '), 5.13 (t, 1H, J
3,4, J
4,59.7Hz, H-4), 5.06 (d, 1H, J
1 ', 2 '1.4Hz, H-1 '), 4.39 (m, 1H, H-3), 4.33-4.26 (m, 2H, H-5, H-5 '), 4.24 (dd, 1H, J
1,21.7Hz, J
2,33.4Hz, H-2), 4.19 (dd, 1H, J
1 ', 2 '2.0Hz, J
2 ', 3 '3.8Hz, H-2 '), 4.15-4.08 (m, 2H, 2 × H-6 '), 3.10 (d, 1H, J8.8Hz, OH), 2.12,2.09,2.05 (3s, 9H, 3 × CH
3cO), 1.27 (d, 1H, J 6.2Hz, H-6).
13c NMR (75MHz, CDCl
3): δ 170.7,169.9,169.5 (3 × COCH
3), 166.9 (COPh), 155.2,150.2,117.6 (2), 114.7 (2) (C
6h
4oCH
3), 97.4,96.3 (2 × C-1), 55.6 (C
6h
4oCH
3), 20.6 (2), 20.5 (3 × COCH
3), 17.5 (C-6).
Synthesizing of embodiment 9, trisaccharide 14
By compound 12 (2.9g, 4.2mmol), compound 13 (2.9g, 4.7mmol) and
molecular sieve (3g) is dissolved in 100mL methylene dichloride; ice bath is cooled to-10 ℃; under nitrogen protection, stir 0.5 hour; add TMS triflate 54 μ L (0.3mmol); keep-10 ℃ of reactions 0.5 hour; remove ice bath and make reactant naturally be warmed up to room temperature, after 2 hours, add 100 μ L triethylamines and finish reaction.Suction filtration reaction solution, concentrated under filtrate decompression, column chromatography for separation (sherwood oil: toluene: ethyl acetate=4: 4: 1, v/v/v) must white foam shape solid disaccharide 14 (3.6g, 75%).[α]
D 25+101.8°(c?0.5CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.04-7.17 (m, 20H, Bz-H), 7.08-7.04 (m, 2H, Ar-H), 6.88-6.84 (m, 2H, Ar-H), 5.62-5.60 (m, 2H, 2 × H-3), 5.60-5.49 (m, 5H, H-1, H-2,3 × H-4), 5.34 (br, 1H, H-1), 5.16 (br, 1H, H-1), 4.75-4.55 (m, 2H, 2 × H-2), 4.50-4.00 (m, 6H, H-3,3 × H-5,2 × H-6), 3.80 (s, 1H, C
6h
4oCH
3), 2.11,2.10,2.06 (3s, 9H, 3 × CH
3cO), 1.42 (d, 1H, J 6.2Hz, H-6), 1.29 (d, 1H, J 6.0Hz, H-6).
13c NMR (75MHz, CDCl
3): δ 170.4,170.0,170.5 (3 × COCH
3), 165.9,165.2,164.9,164.5 (4 × COPh), 155.4,150.2,117.8 (2), 114.7 (2) (C
6h
4oCH
3), 98.7,96.3,95.8 (3 × C-1), 55.6 (C
6h
4oCH
3), 20.6 (2), 20.4 (3 × COCH
3), 17.7,17.6 (2 × C-6).
Synthesizing of embodiment 10, trisaccharide 15
Trisaccharide 14 (1.5g, 1.3mmol) is dissolved in 100mL methyl alcohol, adds 0.25g palladium/carbon, the air in reaction system is extracted out, be filled with 1atm hydrogen, under room temperature, react, after 4 hours, react completely.Suction filtration reaction solution, concentrated under filtrate decompression.This enriched material is dissolved in 10mL pyridine, add 3mL diacetyl oxide, under room temperature, react, after 12 hours, react completely, by reaction solution concentrating under reduced pressure, revolve steaming with toluene (2 × 10mL) azeotropic, enriched material column chromatography for separation (sherwood oil: ethyl acetate=2: 1) white foam shape solid trisaccharide 15 (1.2g, 80%).[α]
D 25+81.6°(c?0.7CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.04-7.17 (m, 20H, Bz-H), 7.08-7.05 (m, 2H, Ar-H), 6.88-6.84 (m, 2H, Ar-H), 5.91 (d, 1H, J8.2Hz, NH), 5.68 (dd, 1H, J
2,33.1Hz, J
3,410.2Hz, H-3), 5.62-5.48 (m, 5H, H-1, H-2-Rhap, H-3,2 × H-4), 5.35-5.25 (m, 2H, H-1, H-4), 5.18 (br, 1H, H-1), 4.77 (m, 1H, H-2-Manp), 4.67-4.38 (m, 3H, H-2, H-3, H-5), 4.38-4.03 (m, 4H, H-5,2 × H-6), 2.14,2.03,2.01 (3s, 9H, 3 × CH
3cOO), 2.08 (s, 3H, CH
3cONH), 1.41 (d, 1H, J5.9Hz, H-6-Rhap), 1.29 (d, 1H, J6.0Hz, H-6-Rhap).
13c NMR (75MHz, CDCl
3): δ 170.4,170.2,169.5 (3 × COCH
3), 165.8,165.2,165.0,164.5 (4 × COPh), 155.2,150.0,117.6 (2), 114.7 (2) (C
6h
4oCH
3), 98.8,98.1,96.0 (3 × C-1), 55.6 (C
6h
4oCH
3), 23.2 (NHCOCH
3), 20.7 (2), 20.6 (3 × COCH
3), 17.7,17.6 (2 × C-6).
Trisaccharide 15 (1.1g, 0.95mmol) is dissolved in the acetonitrile solution of 20mL 80%, adds ceric ammonium nitrate (CAN) (2.1g, 3.8mmol) at 30 ℃, maintain this temperature and react completely after 20 minutes.By reaction solution concentrating under reduced pressure at lower than 50 ℃, methylene dichloride dissolves enriched material, washing, organic phase anhydrous Na
2sO
4dry; concentrated, column chromatography for separation (sherwood oil: ethyl acetate=2: 1; v/v) must take off the trisaccharide that p-methoxyphenyl is protected: 3; 4; 6-tri--O-ethanoyl-2-acetylaminohydroxyphenylarsonic acid 2-deoxidation-α-D-MANNOSE-(1 → 2)-[2; 3,4-, tri--O-benzoyl-α-L-rhamnosyl-(1 → 3)-] 4-O-benzoyl-α-L-rhamnosyl.This trisaccharide (0.90mmol) is dissolved in 40mL anhydrous methylene chloride; under nitrogen protection, add Trichloroacetonitrile (0.5mL, 5mmol) and DBU (0.06mL, 0.4mmol); stirring reaction under room temperature, reacted completely after 2 hours.By reaction solution concentrating under reduced pressure, enriched material column chromatography for separation (sherwood oil: ethyl acetate=2: 1) obtain white foam shape solid trisaccharide III (0.75g, 68%).[α]
D 25+98.3°(c?0.6CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.72 (s, 1H, CNHCCl
3), 8.04-7.17 (m, 20H, Bz-H), 6.41 (d, 1H, J
1.21.6Hz, H-1), 5.85 (d, 1H, J 8.2Hz, NH), 5.75-5.45 (m, 6H, H-1, H-2,2 × H-3,2 × H-4), 5.33 (t, 1H, J9.9Hz, H-4), 5.23 (br, 1H, H-1), 4.80 (m, 1H, H-2-Manp), 4.70 (m, 1H, H-3), 4.57-4.53 (m, 2H, H-2, H-5), 4.50-4.00 (m, 4H, 2 × H-5,2 × H-6), 2.15 (s, 3H, CH
3cOO), 2.10 (s, 3H, CH
3cONH), 2.04 (2) (s, 6H, 2 × CH
3cOO), 1.35 (d, 1H, J 6.5Hz, H-6-Rhap), 1.33 (d, 1H, J6.3Hz, H-6-Rhap).
By Compound I I (190mg, 0.27mmol), compound III (390mg, 0.33mmol) and
molecular sieve (0.5g) is dissolved in 20mL methylene dichloride; ice bath is cooled to-10 ℃; under nitrogen protection, stir 0.5 hour; add TMS triflate 9 μ L (0.05mmol); keep-10 ℃ of reactions 0.5 hour; remove ice bath and make reactant naturally be warmed up to room temperature, after 2 hours, add 15 μ L triethylamines and finish reaction.Suction filtration reaction solution, concentrated under filtrate decompression, column chromatography for separation (sherwood oil: ethyl acetate=1: 2, v/v) obtains white foam shape solid pentasaccharides IV (220mg, 69%).[α]
D 25+50.3°(c?0.5CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, CDCl
3): δ 8.07-8.03 (m, 4H, Bz-H), 7.91 (d, 2H, J 7.4Hz, Bz-H), 7.81 (d, 2H, J 7.3Hz, Bz-H), 7.69 (d, 2H, J 7.6Hz, Bz-H), 7.55-7.27 (m, 14H, Bz-H), 7.19 (t, 2H, J 7.7Hz, Bz-H), 6.91 (d, 2H, J 9.1Hz, Ar-H), 6.78 (d, 2H, J9.1Hz, Ar-H), 5.90-5.85 (m, 2H, 2 × NHAc), 5.64 (dd, 1H, J
2,33.0Hz, J
3,410.2Hz, H-3-RhapC), 5.60 (t, 1H, J
2,3, J
3,49.4Hz, H-3-Glup), 5.55-5.39 (m, 5H, H-2-RhapA, H-2-RhapC, H-3-ManpC, H-4-RhapB, H-4-RhapC), 5.34 (d, 1H, J 1.6Hz, H-1-RhapA), 5.29 (t, 1H, J
3,4, J
4,59.6Hz, H-4-Manp), 5.21 (s, 1H, H-1-Rhap), 5.14 (t, 1H, J
3,4, J
4,59.6Hz, H-4-RhapA), 5.08 (d, 1H, J 8.2Hz, H-1-Glup), 5.05 (s, 1H, H-1-Manp), 4.92 (s, 1H, H-1-Manp Rhap), 4.90-4.70 (m, 3H, H-2-Manp, H-2-RhapB, H-3-RhapB), 4.60-4.25 (m, 6H, H-3-RhapA, 2 × H-6-Manp, 3 × H-5), 4.17 (m, 1H, H-2-Glup), 4.10-3.80 (m, 3H, H-4-Glup, H-5-Glup, H-6-Glup), 3.80-3.70 (m, 4H, H-6-Glup, C
6h
4oCH
3), 3.63 (m, 1H, H-5), 2.17-2.04 (m, 21H, 7 × CH
3cO), 1.94 (s, 3H, CH
3cO), 1.37 (d, 3H, J 6.1Hz, H-6-Rhap), 1.23 (d, 3H, J 5.7Hz, H-6-Rhap), 1.16 (d, 3H, J 6.2Hz, H-6-Rhap).
13c NMR (75MHz, CDCl
3): δ 171.1,170.3,170.2 (3), 170.1,169.7 (2) (8 × CH
3cO), 166.1,165.8,165.2,164.9,164.5 (5 × C
6h
5cO), 155.1,149.8,117.7 (2), 114.5 (2) (C
6h
4oCH
3), 100.8 (C-1-Glup), 98.8,98.3,98.2 (2 × C-1), 96.2 (C-1-RhapA), 75.2,73.5,73.2,73.2,72.7,72.2,72.1,71.6,71.3,70.3,69.3,69.2,69.0,68.6,67.7,66.8,65.0,62.5,62.1,55.6,55.5 (C
6h
4oCH
3), 50.7,23.3,23.1,21.1,20.9 (2), 20.7 (2), 20.68 × CH
3cO), 17.7,17.4,17.2 (5 × C-6-Rhap).
Pentasaccharides IV (160mg, 0.066mmol) is dissolved in to saturated NH
3in-MeOH (40mL) solution, stirring reaction under room temperature, reacted completely after 1 week.Sephadex LH-20 (MeOH) column purification obtains white foam shape solid target pentasaccharides I (79mg, 89%).[α]
D 25-39.5°(c?0.3CHCl
3).
Structural identification data are as follows:
1h NMR (300MHz, MeOD): δ 6.95 (d, 2H, J8.4Hz, Ar-H), 6.83 (d, 2H, J8.2Hz, Ar-H), 5.28 (s, 1H, H-1), 5.04 (s, 1H, H-1), 4.97 (s, 1H, H-1), 4.85 (s, 1H, H-1), 4.61 (d, 1H, J 8.0Hz, H-1-Glup), 4.26-4.24 (m, 2H), 4.00-3.82 (m, 6H), 3.80-3.51 (m, 16H), (3.50-3.30 m, 3H), 1.95,1.94 (2s, 6H, 2 × COCH
3), 1.25-1.15 (m, 6H, 2 × H-6), 1.11 (d, 1H, J5.9Hz, H-6).
13c NMR (75MHz, MeOD): δ 174.8,174.7 (2 × COCH
3), 154.6,149.3,118.7 (2), 115.1 (2) (C
6h
4oCH
3), 102.8 (C-1-Glup), 101.7,98.8,97.2,97.0 (4 × C-1), 80.1,77.5,75.0,74.8,74.6,72.8,72.3,72.1,71.9,70.7,70.2,69.9,69.8,69.7,69.6,69.0,68.7,66.4,60.4,60.2,56.1,55.8,52.8 (C
6h
4oCH
3), 22.2,21.9 (2 × COCH
3), 16.7,16.6,16.4 (3 × C-6) .HRMS for C
41h
65n
2o
24(M+H)
+969.3927.Found:969.3909.
Amino pentasaccharides Compound I indoor plant growth regulating-activity adopts the biological examination method of Wheat coleoptile method segment to measure, test method method in conjunction with (1985) such as William on the basis of (1955) classical ways such as Nitsch is also improved, and concrete grammar is as follows:
Get the Wheat coleoptile of cultivating in the dark 3d, with blade, accurately cut the about 4.0mm section of middle portion and make examination material.It is 0.01,0.1 and the test solution of 1.0mg/L that tested compound is made into concentration with phosphoric acid one citrate buffer solution (pH=5) of 0.01mol/L, with the indolylacetic acid (IAA, purchased from Sigma company, analytical pure) of same concentrations for contrast medicament.Pipette the culture dish (ware is put 1 filter paper) that liquid 3mL is placed in diameter 6.0cm, in each ware, put into 10 sections of Wheat coleoptiles (operation is carried out in the dark), each processing repeats 4 times, under 25 ℃ of constant temperature, in dark, cultivate after 24h, measure the length (mm) of each segment, calculating mean value, according to formula below calculate different concns medicament to promotion rate (%), the results are shown in Table 1:
Promotion rate (%)=(chemicals treatment bud scale mean length-4.0 after 24h)/(contrasting bud scale mean length-4.0 after 24h) × 100
The test-results of the amino pentasaccharides Compound I of table 1 to wheat coleoptile segment elongation
Note: contrasting bud scale mean length after 24h is 6.47mm.
Table 1 result shows, amino pentasaccharides Compound I has preferably Promoting plant growth activity, shows stronger Wheat coleoptile and extend and promote actively within the scope of test concentrations, and when concentration reaches 0.1mg/L, its promotion ability is better than contrasting medicament indolylacetic acid.
The field test that amino pentasaccharides Compound I regulates cotton growth in 2011 in the China Agricultural University of Haidian District, Beijing City village experiment centre carry out, test kind is the new cotton 99B of Insect Resistant Cotton.At cotton growth cotyledon period, carry out foliar spray processing, establish the amino pentasaccharides I of 5mg/L, the contrast of 40mg/L gibberic acid medicament, three processing of clear water contrast, each processing arranges 50 strains and repeats, and 7d " Invest, Then Investigate " seedling height, main root length, the results are shown in Table 2.
The amino pentasaccharides Compound I of table 2 regulates test-results to Cotton Seedling-Growth
Table 2 result shows, amino pentasaccharides Compound I can promote Cotton Seedling-Growth, and being mainly manifested in plant height increases, and the gibberic acid of effect and suitable concentration is suitable; On the other hand, amino pentasaccharides Compound I can promote the growth of root system, shows the physiological effect that gibberic acid does not have.
Embodiment 16, field test
Amino pentasaccharides Compound I regulates the field test of wheat growth to carry out at Haidian District, Beijing City China Agricultural University research park in 2011, and soil fertility level is higher.Experimental cultivar is capital winter No. 8.Clear water contrast and amino pentasaccharides 20mg/L seed soaking are established in test.Sowing on September 28, before surviving the winter, (November 15) sampling survey main stem leaf age, individual plant tiller number, the above tiller number of first three leaf of surviving the winter, the total radical of individual plant, the results are shown in Table 3.
The amino pentasaccharides Compound I of table 3 regulates test-results to wheat growth
Table 3 result shows, amino pentasaccharides Compound I is carried out wheat seed processing, can promote wheat seedling growth before the winter, show increase main stem leaf age, increase the above tiller number of individual plant first three leaf of tiller number, particularly winter, increase the total root number of individual plant simultaneously, be conducive to cultivate strong sprout before the winter, ensure safety and survive the winter.
Claims (9)
1. the compound shown in formula I:
2. the preparation method of compound shown in claim 1 Chinese style I, comprises the steps:
1) three sugar compounds donors shown in the disaccharide compound acceptor shown in formula II and formula III are carried out to condensation reaction, obtain the amino pentasaccharides compound of the full guard shown in formula IV;
Wherein, Ac represents that ethanoyl, Bz represent benzoyl;
2) make the amino pentasaccharides compound of the full guard shown in formula IV carry out protecting group in saturated methanol ammonia solution and remove reaction, obtain compound shown in formula I.
3. method according to claim 2, it is characterized in that: described in step 1), condensation reaction is carried out under the katalysis of TMS triflate, the mol ratio of three sugar compounds donors shown in described TMS triflate and formula III is 0.01-0.5:1; The mol ratio of three sugar compounds donors shown in the disaccharide compound acceptor shown in formula II and formula III is 1:1-1.5.
4. it is characterized in that according to the method in claim 2 or 3: described in step 1), the reaction medium of condensation reaction is methylene dichloride; 20 ℃-0 ℃ of the temperature of reaction Ke Wei – of described condensation reaction, the reaction times is 2-4 hour.
5. according to the method in claim 2 or 3, it is characterized in that: step 2) described in reaction temperature of reaction be 20 ℃-30 ℃, the reaction times is 4-8 days.
6. according to the method in claim 2 or 3, it is characterized in that: the preparation method of the disaccharide compound acceptor shown in described formula II is as follows: take the glucose tribromo-acetyl imines ester shown in formula 1 as glycosyl donor, acyl group rhamnoside shown in formula 2 is glycosyl acceptor, carry out condensation reaction and obtain the 1 → 3-β-connection disaccharide shown in formula 3, described disaccharide obtains the compound shown in formula 4 through deacetylation, compound shown in formula 4 is through 4, 6-hydroxyl isopropylidene obtains the compound shown in formula 5, compound hydrazinolysis shown in formula 5 obtains the compound shown in formula 6, compound acetylize shown in formula 6 obtains the compound shown in formula 7, compound solution fork base protection shown in formula 7 obtains the compound shown in formula 8, then the compound shown in formula 8 is carried out to selectivity 6-position benzoylation and obtain the disaccharide compound acceptor shown in formula II,
Wherein, Ac represents that ethanoyl, Bz represent that benzoyl, Phth represent that phthaloyl, PMP represent p-methoxyphenyl.
7. according to the method in claim 2 or 3, it is characterized in that: the preparation method of three sugar compounds donors shown in described formula III is as follows: take the seminose tribromo-acetyl imines ester shown in formula 9 as glycosyl donor, acyl group rhamnoside shown in formula 10 is glycosyl acceptor, carry out condensation reaction and obtain the 1 → 2-α-connection disaccharide shown in formula 11, the de-allyloxycarbonyl of described disaccharide obtains the disaccharide shown in formula 12; Disaccharide shown in formula 12 obtains with the acyl group rhamnosyl tribromo-acetyl imines ester condensation shown in formula 13 trisaccharide that (1 → 2) shown in formula 14-[α-(1 → 3)-] α-L-is connected, this trisaccharide obtains the trisaccharide shown in formula 15 through nitrine reduction, acetylize, by de-the trisaccharide shown in formula 15 p-methoxyphenyl, then activation, obtain three sugar compounds donors shown in formula III;
Wherein, Ac represents that ethanoyl, Bz represent that benzoyl, Alloc represent that allyl oxygen formyl radical, PMP represent p-methoxyphenyl.
8. compound shown in claim 1 Chinese style I is in the application of preparing in plant-growth regulator.
9. application according to claim 8, is characterized in that: described plant-growth regulator has following at least one effect: 1) promote root growth, 2) promote growth of seedling, 3) increase individual plant total root number, 4) increase individual plant and tiller, 5) cultivate strong sprout.
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| Title |
|---|
| Evelina L. Zdorovenko et al..Structure of the O-polysaccharides of the lipopolysaccharides Structure of the O-polysaccharides of the lipopolysaccharides ATCC 19655 containing two O-methylated monosaccharides.《Carbohydrate Research》.2009,第344卷第2519-2527页. |
| Highly efficient removal of allyloxycarbonyl (Alloc) function provides a practical orthogonal protective strategy for carbohydrates;Zong Guang Hui et al.;《Chinese Chemical Letters》;20091231;第20卷;第127-130页 * |
| Practical preparation of 2-azido-2-deoxy-β-D-mannopyranosyl carbonates and their application in the synthesis of oligosaccharides;Zhang Jianjun et al.;《Carbohydrate Research》;20070907;第342卷;第2810-2817页 * |
| Structure of the O-polysaccharides of the lipopolysaccharides Structure of the O-polysaccharides of the lipopolysaccharides ATCC 19655 containing two O-methylated monosaccharides;Evelina L. Zdorovenko et al.;《Carbohydrate Research》;20091231;第344卷;第2519-2527页 * |
| Synthesis of α-D-GlcpNAc-(1→2)-[α-D-ManpNAc-(1→3)-]α-L-Rhap-(1→2)-α-L-Rhap-(1→3)-α-L-Rhap,the repeating unit of O10 antigen from Acinetobacter baumannii;Zhang Jian Jun et al.;《Chinese Chemical Letters》;20081231;第19卷;第416页 * |
| The structure of the O-antigenic chain of the lipopolysaccharide of Rhizobium trifolii 4s;Wang Y et al.;《Carbohydrate Research》;19941231;第260卷;第315页 * |
| Wang Y et al..The structure of the O-antigenic chain of the lipopolysaccharide of Rhizobium trifolii 4s.《Carbohydrate Research》.1994,第260卷第315页. |
| Zhang Jianjun et al..Practical preparation of 2-azido-2-deoxy-β-D-mannopyranosyl carbonates and their application in the synthesis of oligosaccharides.《Carbohydrate Research》.2007,第342卷第2810-2817页. |
| ZhangJianJunetal..Synthesisofα-D-GlcpNAc-(1→2)-[α-D-ManpNAc-(1→3)-]α-L-Rhap-(1→2)-α-L-Rhap-(1→3)-α-L-Rhap the repeating unit of O10 antigen from Acinetobacter baumannii.《Chinese Chemical Letters》.2008 |
| Zong Guang Hui et al..Highly efficient removal of allyloxycarbonyl (Alloc) function provides a practical orthogonal protective strategy for carbohydrates.《Chinese Chemical Letters》.2009,第20卷第127-130页. |
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