CN104311683A - Synthesis and application of guar gum phosphate ester with high substitution degree - Google Patents
Synthesis and application of guar gum phosphate ester with high substitution degree Download PDFInfo
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- CN104311683A CN104311683A CN201410523569.6A CN201410523569A CN104311683A CN 104311683 A CN104311683 A CN 104311683A CN 201410523569 A CN201410523569 A CN 201410523569A CN 104311683 A CN104311683 A CN 104311683A
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Abstract
The invention discloses a method for synthesizing guar gum phosphate ester with a high substitution degree, belonging to the technical field of biomacromolecule and chemical synthesis. According to the method, a special phosphorylation agent and a microwave auxiliary technique are adopted to efficiently synthesize the guar gum phosphate ester with the high substitution degree and biological activity, so that the application range of the guar gum phosphate ester as a natural active substance is widened. The activity experiment shows that the guar gum phosphate ester synthesized by using the method has a remarkable function of in-vitro HepG2 cell inhibition on HepG2 liver cancer cells (the IC50 is 19.71-28.42mg/mL), can be used as an inhibitor for HepG2 liver cancer cells and can be used for preparing anti-cancer medicines.
Description
Technical field
The invention belongs to biopolymer and chemosynthesis technical field, relate to a kind of guar gum phosphate synthesis method; The present invention also relates to the medicinal use of guar gum phosphoric acid ester simultaneously---and the inhibitor as HepG2 liver cancer cell is applied to be prepared in cancer therapy drug.
Background technology
Polysaccharide is the natural high moleculer eompound that a large class of nature existence is formed by connecting by glycosidic link by monose.There is significant pharmacodynamic feature, as strengthened immunologic function, the biological activity such as antitumor, hypoglycemic, anti-ageing, antiviral and become the focus of research.The biological activity that polysaccharide possesses and its chemical structure have extremely close relation.The textural factor affecting polysaccharide biologic activity comprise molecular weight, with functional group, the main chain character (sugar unit composition and glycosidic link type) of polysaccharide, the type of side chain, monose mode of connection and polysaccharide molecule higher structure as space conformation etc.Wherein the kind of functional group is very big on the bioactive impact of polysaccharide, and the kind as the position of substitution, content and the complexing element of the sulfate on sugar chain, ethanoyl, alkyl etc. determines polysaccharide for what active and active size.Therefore, in order to improve the biological activity of polysaccharide, by changing the space structure of polysaccharide, molecular weight and substituting group kind, number and position, its activity is had an impact.Utilize the hydroxyl on polysaccharide residue radical, carboxyl, amino groups, use chemical process to carry out transformation to the functional group of polysaccharide and modify, become the important directions of active polysaccharide research and screening.
In many polysaccharide derivates, polysaccharide phosphate ester is paid close attention to widely because it has good biologic activity.Bibliographical information is constantly had to synthesize polysaccharide phosphate ester by chemical process in recent years.Suzuki etc. to carry out after stearoylation and phosphorylation yeast mannans can effective anti-S-180 and ehrlich carcinoma (Chemical & pharmaceutical bulletin, 1976,24,1100-1103); Williams etc. are right
βafter-(1-3)-D-dextran has carried out phosphorylation, activity has improved greatly (Carbohydrate Research, 1991,219,203-213); The N of the synthesis such as Chen Ruyu, the anti-tumor activity experimental result of N-bis-(2-chloroethyl) fructosyl phosphoramidate shows, it has certain restraining effect (SCI, 1993,7,963) to L1210 cell and S180 ascites carcinoma.At present in the phosphorylation of polysaccharide, use phosphorylation agent mainly phosphoric acid salt, acid anhydrides etc., directly derivatize is carried out to polysaccharide, there is long reaction time, the shortcoming that substitution value is generally lower.For the polyose biopolymer that molecular weight is larger, due to sterically hindered too large, polysaccharide phosphate ester substitution value prepared by ordinary method is very low, often needs to repeat to react just to obtain the higher product of substitution value.
Guar gum (or guar gum), numbering: GB20.025, remove the peel the endosperm fraction after degerming by the seed of leguminous plants guar-bean (Cyamposis tetragonolobus), add water after drying and crushing, carry out the alcohol settling with 20% after pressurized hydrolysis, drying and crushing after centrifugation and obtaining.Main component to be molecular weight be 5 ~ 800,000 the polygalactomannan that combines of glycosidic linkage, the high molecular hydrocolloid polyose be namely made up of semi-lactosi and seminose (1:2).White is to khaki powder, and can be dispersed in water formation viscous fluid, be natural gum medium viscosity soprano.Its aqueous solution is neutral, and viscosity changes with the change of pH value, and pH value 6 ~ 8 viscosity is the highest, and more than pH10 then reduces rapidly, reduces in pH6 ~ 3.5 with pH value.Below pH3.5 viscosity increases again.Because the abundant aquation of guar gum cumulative in cold and hot water forms translucent solution.Be mainly used in food processing field: 1, in cold drink is as ice-creams, ice cream, frost, borneol, play good stable agent effect, prevent ice crystal from producing, play thickening, emulsifying effect; 2, play in flour products is as noodles, vermicelli, instant noodles, vermicelli and prevent bonding, water conservation, increase biceps, keep the excellent effect of quality, and extended the upper shelf time; 3, in beverage is as Semen arachidis hypogaeae milk, almond milk, Manufacture of Walnut Milk, orange granule juice, fruit juice, fruit tea, various solid beverage and eight treasure gruel, play thickening water holding and stabilizer function, and improve mouthfeel effect; If 4, play stabilizer function in milk-product milk, Yoghourt, and play thickening, emulsification, improve the effect of mouthfeel; 5, in bean product are as bean curd, soymilk, stabilization is played; 6, in meat product, as played bonding, tasty and refreshing and increase volumetric in ham sausage, luncheon meat, various burger; 7, seasonings: in sauce and salad seasonings, make use of guar gum and produces this essential property of high viscosity at low concentrations, makes the matter structure of these products and flows the organoleptic quality high-qualitys more such as change; 8, tinned pre-: the feature of this series products is that guar gum then can be used for the moisture in multiviscosisty product as far as possible not containing the water of flowable state, and makes the gravy of meat dish solid part surface bag one deck stiff.Special, slowly swelling guar gum can also be used for viscosity when limiting tinning sometimes.In order to expand the application of guar gum, its phosphorylation being obtained there is bioactive guar gum phosphoric acid ester tool and being of great significance.
Summary of the invention
The object of this invention is to provide a kind of preparation method of guar gum phosphoric acid ester;
Another object of the present invention is to provide a kind of medicinal use of guar gum phosphoric acid ester---as the inhibitor of HepG2 liver cancer cell for the preparation of cancer therapy drug.
One, the synthesis of polysaccharide phosphate ester
(1) preparation of guar gum solution: joined by guar gum in dry formamide, stirs 12 ~ 24 hours, obtains homogeneous guar gum solution at 70 ~ 80 DEG C; The bulking value of guar gum and dry formamide is 0.01 ~ 0.05g/mL.
(2) preparation of phosphorylation agent: at 0 ~ 5 DEG C, is added drop-wise to phosphorus oxychloride (speed with 0.5 ~ 0.8 mL/ minute) in anhydrous pyridine, reacts 2 ~ 4 hours, obtain phosphorylation agent; The volume ratio of phosphorus oxychloride and anhydrous pyridine is 1:5 ~ 1:15.
In order to fast reaction speed, in the preparation of phosphorylation agent, also add catalyzer DMAP, and the weightmeasurement ratio of DMAP and phosphorus oxychloride is 0.02 ~ 0.05g/mL.
(3) phosphorylation reaction: at 0 ~ 5 DEG C, guar gum solution is added drop-wise in phosphorylation agent, at 40 ~ 80 DEG C, stirring reaction 15 ~ 90 minutes, is cooled to 0 ~ 5 DEG C, after being neutralized to pH=7 ~ 8 with the lye solution of 0 ~ 5 DEG C, suction filtration under room temperature, 0.06 ~ 0.08MPa; It is in 8000 ~ 12000 daltonian dialysis tubings that filtrate loads the molecular weight that dams, and prior to dialysing 36 ~ 48 hours in flowing water, then dialyses 18 ~ 36 hours in deionization; Then slow down liquid pressure-reducing in dialysis tubing and be concentrated into 1/20 ~ 1/40 of original volume, finally add dehydrated alcohol and carry out precipitating (ethanol content accounts for 70 ~ 80% of cumulative volume), centrifugal, lyophilize, obtains guar gum phosphoric acid ester.
The volume ratio of guar gum solution and phosphorylation reagent is 1:2 ~ 1:5; Concentrating under reduced pressure be 50 ~ 60 DEG C, carry out under 0.06 ~ 0.08Mpa; Centrifugal is be separated 15 ~ 25 minutes under 3500 ~ 4500 revs/min; Lyophilize is-60 ~-50 DEG C, lyophilize 30 ~ 36 hours under the condition of 1 ~ 5Pa.
In order to accelerate the substitution value of phosphorylation process and raising phosphoric acid ester, the phosphorylation of guar gum is carried out under the microwave condition of power 100 ~ 500W, frequency 2450MHz.
Two, the structure properties of guar gum phosphoric acid ester
Below by infrared spectrogram,
13c NMR spectrogram, phosphorus detection, molecular weight and distribution thereof and analytic explanation is carried out to the properity etc. of restraining effect to the high substitution value polysaccharide phosphate ester that the present invention synthesizes of HepG2 liver cancer cell.
1, infrared spectra map analysis
Fig. 1 is the infrared spectrogram of guar gum phosphoric acid ester prepared by guar gum and the present invention.In Fig. 1, the middle sugared ring stem absorption peak of guar gum and guar gum phosphoric acid ester does not change.At 1268.9 cm
-1with 909.6 cm
-1the new absorption peak that place occurs, the characteristic absorbance of corresponding P=O and C-O-P, illustrates that the part-OH in guar gum structure is replaced by phosphate radical, by Phosphation respectively.
2,
13c NMR spectrum analysis
Fig. 2 is the guar gum phosphoric acid ester (B) prepared of guar gum (A) and the present invention
13c NMR spectrogram.The main change of guar gum phosphoric acid ester prepared by the present invention exists
δ67.4 and 65.2
δthe chemical shift appearance that 67.4 places are new, is attributed to mannosyl units and the galactose units of C-6 replacement.From
δthe chemical shift at 60 ~ 70 places is seen,
δthe chemical shift of about 60 C-6 does not change.Can infer thus, there occurs C-6 in guar gum phosphoric acid ester and replace.
3, phosphorus detection
Adopt inductively coupled plasma atomic emission spectrometry (ICP-AES) to measure phosphorus content, in the present invention, the substitution value of the guar gum phosphoric acid ester of regio-selective synthesis is 0.88 ~ 1.37, illustrates that polysaccharide phosphate ester successfully synthesizes.
4, molecular weight analyse
Adopt gel chromatography-dynamic static light scattering coupling to measure, in the present invention, the weight-average molecular weight of the guar gum phosphoric acid ester of regio-selective synthesis is 6.08 × 10
4da ~ 8.23 × 10
4da, molecular weight distribution is 2.76 ~ 3.31.Illustrate that the molecular weight of polysaccharide phosphate ester in the present invention can regulate according to reaction conditions.
5, the external restraining effect to HepG2 liver cancer cell
The HepG-2 cell of taking the logarithm vegetative period, uses 0.25% tryptic digestion, and makes concentration 3 × 10 with the DMEM substratum containing 15% new-born calf serum
5the single cell suspension of individual/mL, constantly shake cell suspension makes cell count even, is inoculated in 96 well culture plates (panel edges envelope sterilized water, blank group adds the not celliferous substratum in 3 holes), puts CO with 0.2 ml/ hole
2inherent 37 DEG C of incubator, saturated humidity, 5%CO
2cultivate under condition.Cultivate cell attachment after 24 h, change pastille substratum, every hole adds 190 μ L substratum and 10 μ L sample liquid (blank group and control group add 10 μ l PBS liquid), and medicine final concentration is 20,50,100,200,500,1000,5000 μ g/mL, every concentration repeats 6 holes, and establishes the control group and blank zeroing group that do not add sample.Put CO
2in incubator, 37 DEG C, saturated humidity, 5%CO
2cultivate under condition.After sample preparation 48 h, every hole adds MTT solution 20 μ L, continues cultivation 4 h at 37 DEG C.Carefully discard supernatant liquor in hole, and add 150 μ L DMSO, vibrate 10 min, and purple crystal is dissolved completely.Survey light absorption values by microplate reader at 490 nm places, control group with the zeroing of blank group, and by following formulae discovery inhibiting rate, and according to the inhibiting rate calculation of half inhibitory concentration (IC of different concns sample
50):
Inhibiting rate (%)=(1-experimental group mean OD value/control group mean OD value) × 100%
To the IC of HepG2 liver cancer cell
50be 19.71 ~ 28.42mg/mL, therefore, it has the effect of significant vitro inhibition HepG2 cell, can be used as the inhibitor of HepG2 liver cancer cell, for the preparation of cancer therapy drug.
In sum, the present invention compared with prior art tool have the following advantages:
1, the present invention has prepared the guar gum phosphoric acid ester with significant vitro inhibition HepG2 cell by melon to your bean gum phosphorylation, has expanded the range of application of guar gum as natural active matter;
2, the present invention introduces catalyzer DMAP in the synthesis of phosphorylation agent, introduces Microwave-assisted synthesis technology, not only effectively shortens reaction required time, and effectively improve the substitution value of guar gum phosphoric acid ester in Phosphorylation events;
3, the substitution value of guar gum phosphoric acid ester prepared of the present invention and molecular weight can by the proportion adjustment optimizations of phosphorylation agent, to meet different field requirement;
4, the present invention does not need specific installation, and simply controlled, cost is low, is applicable to applying.
Accompanying drawing explanation
Fig. 1 is the infrared spectrogram of guar gum phosphoric acid ester prepared by guar gum and the present invention.
Fig. 2 is the guar gum phosphoric acid ester (B) prepared of guar gum (A) and the present invention
13c NMR spectrogram.
Embodiment
Be further described below by the synthetic method of specific embodiment to high substitution value polysaccharide phosphate ester of the present invention.
embodiment 1
(1) preparation of guar gum solution: take 0.5g guar gum, joins in 50mL dry formamide, at 80 DEG C, magnetic stirring apparatus stirs 12 hours with the rotating speed of 1200 revs/min, obtains homogeneous guar gum solution;
(2) preparation of phosphorylation agent: the reactor with condensation, stirring and constant pressure funnel device is placed in 5 DEG C of cryosels baths, first 60mL pyridine is joined in reactor, then the piston of 12mL phosphorus oxychloride with constant pressure funnel is added drop-wise in reactor with 0.8mL/ minute acceleration; Add DMAP 0.6g afterwards, continue reaction 30 minutes, obtain phosphorylation agent;
(3) phosphorylation reaction: the reactor with stirring and constant pressure funnel device is placed in 0 DEG C of cryosel bath, first 50mL phosphorylation agent is joined in reactor, the piston of 10mL guar gum solution with constant pressure funnel is added drop-wise in reactor with 2mL/ minute acceleration; Take out reactor and put into microwave reaction device, be 500W, frequency 2450MHz at microwave power, stirring reaction 60 minutes at 70 DEG C, be cooled to 0 DEG C, pH=7 is neutralized to the sodium hydroxide solution of 0 DEG C, then suction filtration under room temperature, 0.06MPa, it is in 8000 ~ 12000 daltonian dialysis tubings that filtrate loads combined closure system, prior to dialysing 48 hours in flowing water, dialyse 36 hours (within every 2 hours, changing water once) in deionized water again, then by liquid in dialysis tubing 60 DEG C, be evaporated to 1/40 of original volume under 0.08MPa; Add ethanol to carry out precipitating (ethanol content accounts for 80% of cumulative volume), finally under the condition of 4500 revs/min centrifugal 25 minutes, solid ,-60 DEG C, lyophilize 30 hours under the condition of 1Pa, obtains guar gum phosphoric acid ester.
The substitution value of this polysaccharide phosphate ester is 1.37, and weight-average molecular weight is 8.23 × 10
4da, molecular weight distribution is 2.97.To the IC of HepG2 liver cancer cell
50for 23.36mg/mL.
embodiment 2
(1) preparation of guar gum solution: take 2.5g guar gum, joins in 50mL dry formamide, at 75 DEG C, magnetic stirring apparatus stirs 24 hours with the rotating speed of 800 revs/min, obtains homogeneous guar gum solution;
(2) preparation of phosphorylation agent: the reactor with condensation, stirring and constant pressure funnel device is placed in 0 DEG C of cryosel bath, first 60mL pyridine is joined in reactor, then the piston of 4mL phosphorus oxychloride with constant pressure funnel is added drop-wise in reactor with 0.5mL/ minute acceleration; Add DMAP 0.08g afterwards, continue reaction 30 minutes, obtain phosphorylation agent;
(3) phosphorylation reaction: the reactor with stirring and constant pressure funnel device is placed in 5 DEG C of cryosels baths, first 50mL phosphorylation agent is joined in reactor, then the piston of 25mL guar gum solution with constant pressure funnel is added drop-wise in reactor with 1mL/ minute acceleration; Take out reactor and put into microwave reaction device, be 100W, frequency 2450MHz at microwave power, stirring reaction 60 minutes at 40 DEG C, be cooled to 0 DEG C, pH=8 is neutralized to the sodium hydroxide solution of 0 DEG C, suction filtration under room temperature, 0.08MPa, it is in 8000 ~ 12000 daltonian dialysis tubings that filtrate loads combined closure system, prior to dialysing 36 hours in flowing water, dialyse 48 hours (within every 5 hours, changing water once) in deionized water again, then by liquid in dialysis tubing 50 DEG C, be evaporated to 1/20 of original volume under 0.06MPa; Add ethanol to carry out precipitating (ethanol content accounts for 70% of cumulative volume), finally under the condition of 3500 revs/min centrifugal 15 minutes, solid ,-50 DEG C, lyophilize 36 hours under the condition of 5Pa, obtains guar gum phosphoric acid ester.
The substitution value of this polysaccharide phosphate ester is 1.13, and weight-average molecular weight is 6.08 × 10
4da, molecular weight distribution is 3.31.To the IC of HepG2 liver cancer cell
50for 19.71mg/mL.
embodiment 3
(1) preparation of guar gum solution: take 1g guar gum, joins in 50mL dry formamide, at 70 DEG C, magnetic stirring apparatus stirs 18 hours with the rotating speed of 500 revs/min, obtains homogeneous guar gum solution;
(2) preparation of phosphorylation agent: the reactor with condensation, stirring and constant pressure funnel device is placed in 2 DEG C of cryosels baths, first 60mL pyridine is joined in reactor, then the piston of 6mL phosphorus oxychloride with constant pressure funnel is added drop-wise in reactor with 0.7mL/ minute acceleration; Add DMAP 0.3g afterwards, continue reaction 30 minutes, obtain phosphorylation agent;
(3) phosphorylation reaction: the reactor with stirring and constant pressure funnel device is placed in 2 DEG C of cryosels baths, first 50mL phosphorylation agent is joined in reactor, then the piston of 12.5mL guar gum solution with constant pressure funnel is added drop-wise in reactor with 1mL/ minute acceleration; Take out reactor and put into microwave reaction device, be 300W, frequency 2450MHz at microwave power, stirring reaction 15 minutes at 80 DEG C, be cooled to 2 DEG C, pH=7 is neutralized to the sodium hydroxide solution of 2 DEG C, suction filtration under room temperature, 0.07MPa, it is in 8000 ~ 12000 daltonian dialysis tubings that filtrate loads combined closure system, prior to dialysing 48 hours in flowing water, dialyse 18 hours (within every 3 hours, changing water once) in deionized water again, then by liquid in dialysis tubing 55 DEG C, be evaporated to 1/30 of original volume under 0.07MPa; Add ethanol to carry out precipitating (ethanol content accounts for 80% of cumulative volume), finally under the condition of 4000 revs/min centrifugal 15 minutes, solid ,-55 DEG C, lyophilize 36 hours under the condition of 3Pa, obtains guar gum phosphoric acid ester.
The substitution value of this polysaccharide phosphate ester is 0.88, and weight-average molecular weight is 6.30 × 10
4da, molecular weight distribution is 2.76.To the IC of HepG2 liver cancer cell
50for 20.77mg/mL.
embodiment 4
(1) preparation of guar gum solution: take 0.5g guar gum, joins in 50mL dry formamide, at 80 DEG C, magnetic stirring apparatus stirs 12 hours with the rotating speed of 1200 revs/min, obtains homogeneous guar gum solution;
(2) preparation of phosphorylation agent: the reactor with condensation, stirring and constant pressure funnel device is placed in 5 DEG C of cryosels baths, first 60mL pyridine is joined in reactor, the piston of 12mL phosphorus oxychloride with constant pressure funnel is added drop-wise in reactor with 0.8mL/ minute acceleration, react 30 minutes, obtain phosphorylation agent;
(3) phosphorylation reaction: the reactor with stirring and constant pressure funnel device is placed in 0 DEG C of cryosel bath, first 50mL phosphorylation agent is joined in reactor, fill the piston of 10mL guar gum solution with constant pressure funnel and be added drop-wise in reactor by opening with 2mL/ minute acceleration again, stirring reaction 60 minutes at 70 DEG C, be cooled to 0 DEG C afterwards, pH=7 is neutralized to the sodium hydroxide solution of 0 DEG C, room temperature, suction filtration under 0.06MPa, it is in 8000 ~ 12000 daltonian dialysis tubings that filtrate loads combined closure system, prior to dialysing 48 hours in flowing water, 36 hours (within every 2 hours, changing water once) is dialysed again in deionized water, then by liquid in dialysis tubing at 60 DEG C, 1/40 of original volume is evaporated under 0.08MPa, add ethanol to carry out precipitating (ethanol content accounts for 80% of cumulative volume), finally under the condition of 4500 revs/min centrifugal 25 minutes, lower floor's solid ,-60 DEG C, lyophilize 30 hours under the condition of 1Pa, obtains guar gum phosphoric acid ester.
The substitution value of this polysaccharide phosphate ester is 0.54, and weight-average molecular weight is 7.83 × 10
4da, molecular weight distribution is 3.19.To the IC of HepG2 liver cancer cell
50for 28.42mg/mL.
Each parameter of this embodiment is corresponding with embodiment 1, the difference of the two is: embodiment 1 adds catalyzer DMAP in the preparation of phosphorylation agent, in the phosphorylation of guar gum, have employed microwave assisted techniques, but the biological activity of the product obtained there is significant raising.
Claims (10)
1. high substitution value guar gum phosphate synthesis method, comprises following processing step:
(1) preparation of guar gum solution: joined by guar gum in dry formamide, stirs 12 ~ 24 hours, obtains homogeneous guar gum solution at 70 ~ 80 DEG C;
(2) preparation of phosphorylation agent: at 0 ~ 5 DEG C, is added drop-wise to phosphorus oxychloride in anhydrous pyridine, reacts 2 ~ 4 hours, obtains phosphorylation agent;
(3) phosphorylation reaction: at 0 ~ 5 DEG C, guar gum solution is added drop-wise in phosphorylation agent, at 40 ~ 80 DEG C, stirring reaction 15 ~ 90 minutes, is cooled to 0 ~ 5 DEG C, after being neutralized to pH=7 ~ 8 with the lye solution of 0 ~ 5 DEG C, suction filtration under room temperature, 0.06 ~ 0.08MPa; It is in 8000 ~ 12000 daltonian dialysis tubings that filtrate loads the molecular weight that dams, and prior to dialysing 36 ~ 48 hours in flowing water, then dialyses 18 ~ 36 hours in deionization; Then slow down liquid pressure-reducing in dialysis tubing and be concentrated into 1/20 ~ 1/40 of original volume, finally add dehydrated alcohol and precipitate, centrifugal, lyophilize, obtains guar gum phosphoric acid ester.
2. high substitution value guar gum phosphate synthesis method as claimed in claim 1, it is characterized in that: in step (1), the bulking value of guar gum and dry formamide is 0.01 ~ 0.05g/mL.
3. high substitution value guar gum phosphate synthesis method as claimed in claim 1, it is characterized in that: in step (2), the volume ratio of phosphorus oxychloride and anhydrous pyridine is 1:5 ~ 1:15.
4. high substitution value guar gum phosphate synthesis method as claimed in claim 1, it is characterized in that: step (2), in the synthesis of phosphorylation agent, add catalyzer DMAP, and the weightmeasurement ratio of DMAP and phosphorus oxychloride is 0.02 ~ 0.05g/mL.
5. high substitution value guar gum phosphate synthesis method as claimed in claim 1, it is characterized in that: in step (3), the volume ratio of guar gum solution and phosphorylation reagent is 1:2 ~ 1:5.
6. high substitution value guar gum phosphate synthesis method as claimed in claim 1, it is characterized in that: in step (3), phosphorylation reaction carries out under the microwave condition of power 100 ~ 500W, frequency 2450MHz.
7. high substitution value guar gum phosphate synthesis method as claimed in claim 1, is characterized in that: in step (3), concentrating under reduced pressure be 50 ~ 60 DEG C, carry out under 0.06 ~ 0.08Mpa.
8. high substitution value guar gum phosphate synthesis method as claimed in claim 1, is characterized in that: in step (3), and centrifugal is be separated 15 ~ 25 minutes under 3500 ~ 4500 revs/min.
9. high substitution value guar gum phosphate synthesis method as claimed in claim 1, is characterized in that: in step (3), and lyophilize is-60 ~-50 DEG C, lyophilize 30 ~ 36 hours under the condition of 1 ~ 5Pa.
10. the high substitution value guar gum phosphoric acid ester of method synthesis is as claimed in claim 1 applied in the preparation of cancer therapy drug as the inhibitor of HepG2 liver cancer cell.
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| EP3307818A4 (en) * | 2015-06-10 | 2019-03-06 | Rhodia Operations | Phosphonated polysaccharides and gels and process for making same |
| US10844140B2 (en) | 2015-06-10 | 2020-11-24 | Rhodia Operations | Phosphonated polysaccharides and gels and process for making same |
| WO2017118389A1 (en) * | 2016-01-04 | 2017-07-13 | 黄永 | Glycosaminoglycan ester and application thereof |
| US10889608B2 (en) | 2016-01-04 | 2021-01-12 | Chengdu Auli Ecological Technology Development Co., Ltd. | Ester of aminoglycan and uses thereof |
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