Platycodon juncoides polysaccharide, preparation method thereof and application thereof in preparing medicine for treating psoriasis

Abstract

The application discloses jujube chrysanthemum powder polysaccharide, a preparation method thereof and application of the jujube chrysanthemum powder polysaccharide in preparing a medicine for treating psoriasis. The jujube chrysanthemum powder polysaccharide contains fructose and glucose, and the weight average molecular weight is 2-10KD. The jujube chrysanthemum powder polysaccharide has a remarkable anti-psoriasis effect, is safe and non-toxic, has good biocompatibility, and is expected to be developed into a new type of therapeutic medicine for treating psoriasis.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of natural medicine, and particularly relates to a platycodon jujube polysaccharide, a preparation method thereof and application thereof in preparing a medicine for treating psoriasis. BACKGROUND

[0002] Psoriasis is a common chronic immune-mediated proliferative skin disease, and its main skin pathological features include epidermal hyperplasia, obvious keratinocyte proliferation, increased and expanded angiogenesis, and obvious inflammatory infiltration. There are many psoriasis patients, and there are many new cases every year. Psoriasis has a complex etiology, which may be caused by genetic factors, environmental factors and autoimmune factors, etc., but its pathogenesis is not clear at present, so it is stubborn and difficult to cure, and it is easy to relapse, at the same time, it also increases the possibility and incidence of metabolic syndrome and cardiovascular disease, which brings heavy life burden and mental pressure to patients.

[0003] At present, the commonly used clinical treatment drugs mainly include topical external drugs, traditional treatment drugs, biological agents, small molecule targeted drugs, etc. The external drugs include himox, vitamin D3 derivative, geanthrocol, vitamin A acid cream, glucocorticoid, etc.; the traditional treatment drugs include methotrexate, cyclosporine, acitretin, etc.; the biological agents include TNF-alpha inhibitor (etanercept, adalimumab), IL-12 / 23 inhibitor (ustekinumab), IL-23 inhibitor (guselkumab), etc.; the small molecule targeted drugs include PDE4 inhibitor (apilimod), JAK1~3 inhibitor (tofacitinib, upadacitinib), TYK2 inhibitor (deucravacitinib), etc.

[0004] The external drug treatment is easy to accept for patients and has high compliance, but it is invalid for moderate and severe patients; the traditional treatment drug has significant effect and is convenient to use, but it has adverse reactions such as liver and kidney toxicity, high blood pressure and high blood fat, and the effective rate of the first-line drug methotrexate is only 30%-45%; the biological agent has good effect, but the health status of the patient needs to be fully evaluated before treatment, and the long-term effect and safety need to be observed due to short clinical application time; the small molecule targeted drug has great difference in the effect and safety in treating psoriasis, and there is a risk of serious adverse reactions. Therefore, it is an urgent need to find a new safe and effective treatment drug for psoriasis. SUMMARY

[0005] The purpose of the present application is to provide a platycodon jujube polysaccharide, a preparation method thereof and application thereof in preparing a medicine for treating psoriasis.

[0006] The purpose of the present application is achieved by the following technical solutions.

[0007] A platycodon jujube polysaccharide contains fructose and glucose, and the molar concentration ratio of the two is 95.6:4.4.

[0008] The weight average molecular weight of the polysaccharide is 2-10KD, preferably 3-5KD, particularly preferably 3.8KD.

[0009] A total polysaccharide of Platycodon grandiflorum Juss, the main chain of which is fructose, and the structure of which is shown in formula I:

[0010]

[0011] The numbers in the main chain and the side chain represent the connection sites, and the arrow represents the connection direction.

[0012] The weight average molecular weight of the total polysaccharide of Platycodon grandiflorum Juss is 2-10KD;

[0013] In formula I, n1 is 1-15, (n2+n6) is 20-40, n3 is 1-10, and (n4+n5) is 30-90;

[0014] Preferably, the weight average molecular weight of the total polysaccharide of Platycodon grandiflorum Juss is 3-5KD;

[0015] In formula I, n1 is 6-10, (n2+n6) is 25-30, n3 is 2-4, and (n4+n5) is 55-65;

[0016] Particularly preferably, the weight average molecular weight of the total polysaccharide of Platycodon grandiflorum Juss is 3.8KD;

[0017] In formula I, n1 is 8.25, (n2+n6) is 28.53, n3 is 2.25, and (n4+n5) is 60.96.

[0018] The preparation method of the total polysaccharide of Platycodon grandiflorum Juss comprises the following steps:

[0019] (1) Taking Platycodon grandiflorum Juss medicinal material decoction pieces, defatting, then extracting with alkaline water, concentrating the extract, precipitating with ethanol solution, redissolving the precipitate with water, removing protein by Sevage method, and freeze-drying to obtain total polysaccharide of Platycodon grandiflorum Juss;

[0020] (2) Redissolving the total polysaccharide of Platycodon grandiflorum Juss, centrifuging, adding the supernatant to a chromatography column, collecting the eluate, concentrating and dialyzing to obtain the total polysaccharide of Platycodon grandiflorum Juss of the present application;

[0021] The defatting in step (1) uses 70% ethanol solution.

[0022] The ethanol solution in step (1) is preferably 80% ethanol solution.

[0023] The alkaline water for extraction in step (1) is preferably one or more of ammonia water, sodium hydroxide aqueous solution, and sodium bicarbonate aqueous solution.

[0024] The chromatographic column in step (2) is a cellulose anion exchange chromatographic column; preferably a DEAE-52 cellulose anion exchange chromatographic column.

[0025] The specific step of collecting the eluate in step (2) is:

[0026] During the elution process, the equilibrium flow rate is set to 2.5 min / 10 mL, and water, 0.1 mol / L, 0.2 mol / L, 0.4 mol / L, 0.8 mol / L, and 1 mol / L NaCl are used as the mobile phase in sequence for elution. The eluate of the 0.1 M NaCl solution is collected.

[0027] The dialysis in step (2) is dialysis for 3 days by loading a 3000 Da dialysis bag, and the dialysis water is replaced every day.

[0028] Animal experiments show that the platycodon juncen polysaccharide has a significant anti-psoriasis effect, and can be used for preparing a drug for treating psoriasis.

[0029] The drug for treating psoriasis comprises a therapeutically effective amount of the platycodon juncen polysaccharide and a pharmaceutically acceptable carrier, and the carrier is, for example, water, starch, lactose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, and other commonly used pharmaceutical excipients, preservatives such as sodium benzoate and nipagin ethyl, glidants such as magnesium stearate, disintegrants such as sodium carboxymethyl cellulose, and the like.

[0030] The platycodon juncen polysaccharide can be administered to a patient in need in the form of a pharmaceutical composition, which can be prepared into not only liquid preparations such as oral liquids, but also solid preparations such as tablets, capsules, granules, and pills. The administration route can be oral or local administration, for example, transdermal administration. The dosage is 0.1-5 g / day, which can be determined by a doctor according to the specific condition of the patient.

[0031] The present application has the following advantages and effects relative to the prior art:

[0032] The platycodon juncen polysaccharide has a significant anti-psoriasis effect, is safe and non-toxic, has good biocompatibility, and is expected to be developed as a new therapeutic drug for treating psoriasis. BRIEF DESCRIPTION OF DRAWINGS

[0033] Figure 1 It is an elution curve of the DEAE-52 column.

[0034] Figure 2 It is an HPGPC graph of the polysaccharide of the present application.

[0035] Figure 3 It is a monosaccharide composition graph of the polysaccharide of the present application.

[0036] Figure 4Infrared spectrum of the polysaccharide of the present application.

[0037] Figure 5 Total ion chromatogram of the methylated glycol acetyl ester derivative of the polysaccharide of the present application.

[0038] Figure 6 H-NMR spectrum of the polysaccharide of the present application. 1

[0039] Figure 7 C-NMR spectrum of the polysaccharide of the present application. 13

[0040] Figure 8 DEPT spectrum of the polysaccharide of the present application.

[0041] Figure 9 HSQC-NMR spectrum of the polysaccharide of the present application.

[0042] Figure 10 HMBC-NMR spectrum of the polysaccharide of the present application.

[0043] Figure 11 Effect of the polysaccharide of the present application on the imiquimod-induced psoriasis model of BALB / c mice. DETAILED DESCRIPTION

[0044] The present application will be further described in detail below with reference to the examples and the accompanying drawings, but the embodiments of the present application are not limited thereto.

[0045] Example 1

[0046] The preparation method of the Platycodon inulin uniform polysaccharide comprises the following steps:

[0047] (1) The Platycodon is sliced and ground, and is subjected to fat removal by heating circulation with 70% ethanol through a No. 6 sieve. After drying, the defatted medicinal material is obtained. The defatted medicinal material is added with 25 times of ammonia water solution with a pH value of 12, and is extracted by heating circulation for 2 h. After hot filtration, the filtrate is combined and extracted again. The pH value is adjusted to 7 with an acetic acid aqueous solution, and is concentrated under reduced pressure to a crude drug concentration of 0.2 g / mL. The 85% ethanol is used for precipitation, and the centrifugation is performed. The polysaccharide is dissolved with ultrapure water. The polysaccharide is transferred to a 3000 Da dialysis bag for dialysis for 72 h, and the pure water is replaced every 4 h. The polysaccharide is freeze-dried and weighed to obtain the total polysaccharide of the Platycodon.

[0048] (2) 2 g of the dried total polysaccharide of the Platycodon is added with 100 mL of pure water for sufficient dissolution. Then, the centrifugation is performed at a speed of 3000 r / min for 15 min. The supernatant is subjected to DEAE-52 cellulose anion exchange chromatography column for 200 tubes, and each tube is 10 mL. The polysaccharide content is determined by the phenol-sulfuric acid method, and the elution curve is drawn. Figure 1 ​​). The collected solution was concentrated to 50 mL, and then the concentrated polysaccharide solution was placed in a dialysis bag with a molecular weight of 3000 Da, dialyzed for 3 days, and the pure water was changed more than 6 times a day. Finally, the solution in the dialysis bag was freeze-dried, and the part eluted by the 0.1M NaCl solution was the polysaccharide of the application.

[0049] Example 2

[0050] Determination of sugar content and protein content of the polysaccharide of the application

[0051] Accurately weighed 5.0 mg of fructose standard, added ultrapure water to prepare a 1 mg / mL fructose standard solution, and continuously diluted by half to obtain a series of fructose standard solution of 500, 250, 125, 62.5, 31.25 μg / mL. Similarly, accurately weigh 5.0 mg of polysaccharide obtained in Example 1, dissolve with ultrapure water to prepare a 1 mg / mL sample solution, and dilute to 500 μg / ml for standby. The polysaccharide content was detected by phenol-sulfuric acid method, that is, 200 μL of standard or polysaccharide sample solution was added to a clean test tube, 100 μL of 5% phenol solution was added and shaken, 500 μL of concentrated sulfuric acid solution was added and shaken, and after cooling, 150 μL of reaction solution was taken to a 96-well plate, and the absorbance was detected at 490 nm by an enzyme-labeled instrument. The fructose standard curve was drawn with the standard concentration as the abscissa and the absorbance as the ordinate, the linear equation was obtained, and the polysaccharide content was calculated. The protein content of the polysaccharide of the application was measured by BCA kit method.

[0052] The results show that the sugar content of the polysaccharide of the application is 96.58%, and the protein content is not detected.

[0053] Example 3

[0054] Determination of the molecular weight of the polysaccharide obtained in Example 1

[0055] Respectively take 3 mg of dextran standard of 3690, 2370, 1190, 680, 450, 35 and 4 Kd, and prepare a 1.0 mg / mL control solution with ultrapure water, and pass through a 0.45 μm water filter membrane. The average molecular weight log(Mw) of the standard is detected by Agilent 1260 high performance liquid chromatograph (configured with a differential detector), and the standard curve is drawn with the average molecular weight log(Mw) of the standard as the abscissa and the retention time as the ordinate. Chromatographic analysis conditions: use TSKG-5000PWXL chromatographic column, water as mobile phase, column temperature 40℃, flow rate set to 0.4 mL / min, differential refractive index detector temperature also set to 40℃, and sample injection amount to 10 μL. Accurately take 3 mg of the sample, test according to the above procedure, record the retention time, and input it into the standard equation to calculate the molecular weight distribution of the polysaccharide.

[0056] The results show that the polysaccharide of the application is a single symmetrical peak, and the molecular weight is 3800 Da. Figure 2).

[0057] Example 4

[0058] Determination of monosaccharide composition of polysaccharide of the present application

[0059] Preparation of test sample: 5.0 mg of polysaccharide sample obtained in Example 1 was accurately weighed into a 50 mL round-bottom flask, followed by the addition of 4 mL of trifluoroacetic acid solution with a concentration of 0.5 M. The flask was placed in an oil bath, and hydrolysis was carried out at 80°C with stirring for 40 minutes. After the completion of hydrolysis, the reaction solution was transferred to a rotary evaporator for rotary evaporation to dryness. Then 4 mL of chromatographic methanol was added, and rotary evaporation to dryness was carried out again until there was no acid smell, so as to completely remove the residual trifluoroacetic acid.

[0060] Preparation of mixed standard sample: 5 mg of Glc, fructose (Fru), Man, Rha, Gala, Gal, GclA, and Ara standard samples were accurately weighed. Then 5 mL of ultrapure water was added, and the mixture was dissolved to prepare a mixed standard sample solution with a concentration of 30 mg / mL. In order to ensure the accuracy of the experiment, the mixed solution was further diluted to prepare a series of concentration solutions with concentrations of 3000, 1500, 750, 375, 187.5, 93.75 μg / mL, respectively, for standby use.

[0061] 50 μL of the polysaccharide solution obtained in Example 1 and standard sample solutions with different concentrations were accurately pipetted, and 100 μL of p-aminobenzoic acid methanol solution was added. Then, the reaction test tube was placed in a metal bath at 70°C, and heated for 40 minutes. The test tube was taken out and cooled to room temperature. Then, ultrapure water was added to dilute to 2.5 mL, and the sample was mixed well. The treated sample was filtered through a 0.45 μm microporous filter, and then transferred to a liquid chromatography vial for testing. The liquid chromatography conditions were as follows: the column temperature was set to 30°C, the flow rate of the mobile phase was set to 0.8 mL / min, and the sample injection amount was 10 μL. The wavelength of the ultraviolet detector (DAD) was set to 303 nm, and the excitation wavelength and emission wavelength of the fluorescence detector (FLD) were 313 nm and 358 nm, respectively. The mobile phase B was 0.1 M phosphate buffer (pH = 2.54) containing tetrabutylammonium sulfate, and the mobile phase A was a mixture of B and chromatographic methanol in a volume ratio of 1:1. The liquid phase was separated according to the preset gradient elution program: 0-50 min: 5% A→10% A; 50-55 min: 10% A→20% A; 55-59 min: 20% A→50% A; 59-64 min: 50% A; 64-66 min: 50% A→5% A; 66-71 min: 5% A.

[0062] The results show that the polysaccharide of the present application mainly contains fructose and glucose, and the molar concentration ratio is 95.6:4.4. Figure 3 )。

[0063] Example 5

[0064] Infrared spectrum determination of polysaccharide of the present application

[0065] Take polysaccharide 3mg obtained in Example 1, use Fourier transform infrared spectrometer to conduct infrared determination in the range of 4000-400cm -1 .

[0066] The result shows that the polysaccharide of the present application has a wide and deep peak at 3268cm -1 , which indicates the existence of intermolecular O-H vibration, while the peaks at 2930cm -1 and 2882cm -1 correspond to C-H vibration. In addition, the peaks at 1120cm -1 and 1032cm -1 correspond to O-H variable angle vibration and ether bond C-O-O vibration respectively. According to the peaks at 933cm -1 , 870cm -1 and 820cm -1 , it can be inferred that the polysaccharide of the present application contains furanose ring. Figure 4

[0067] Example 6

[0068] Methylation determination of polysaccharide of the present application

[0069] Put 20.0mg of polysaccharide obtained in Example 1 into a 5mL centrifuge tube, and dry at 80℃ for 10min. Then, add 4mL of anhydrous dimethyl sulfoxide (DMSO), replace the air with nitrogen and seal for ultrasonic treatment for 2h, then add 2mL of anhydrous DMSO-NaOH suspension, replace the air in the bottle with nitrogen again, and ultrasonic react at a temperature of 18-20℃ for 1h. Slowly add iodomethane under light shielding conditions, add in three times, 500μL each time, a total of 1.5mL of iodomethane, then continue to ultrasonic react under light shielding at 18-20℃ for 40min. After the reaction is completed, add 1mL of ultrapure water to terminate the reaction, and the solution is dialyzed in a 3000Da dialysis bag overnight. The next day, pour the liquid in the dialysis bag into a separatory funnel, and add an equal amount of dichloromethane for extraction. After standing for 10min, discard the upper water layer, and continue to extract by adding an equal amount of ultrapure water. A total of three extractions are conducted, and the lower organic layer is poured into a clean glass centrifuge tube, and dried to obtain a powder sample. Infrared detection 3700-3200cm -1 ​hydroxyl functional groups were reacted completely. The methylated sample was completely dried, 2 mL of 88% formic acid was added accurately, and after being dissolved completely, it was transferred to a dry and clean round bottom flask. Then, nitrogen was replaced to ensure that the reaction environment was inert, and it was reacted in a 100°C oil bath pot for 4h. After the reaction was completed, the flask was taken out and naturally cooled to room temperature. A rotary evaporator was used to spin dry the reaction liquid at 45°C, then 3 mL of chromatographic methanol was added and spin dried until the acid smell was completely removed to obtain the treated sample. The sample obtained in the previous step was taken, 3 mL of ultrapure water was added to dissolve it, 0.5 mL was taken for subsequent reaction, 1.5 mL of ultrapure water was added, 2 mL of 4M trifluoroacetic acid solution (the final reaction liquid is 2M trifluoroacetic acid), and it was reacted at 80°C for 4h. After cooling, the rotary evaporator was used to spin dry under reduced pressure (45°C), and chromatographic methanol was added to spin dry to remove the acid. The sample obtained in the previous step was taken, and 1.5 mL of 50mM NaOH solution was added to dissolve it. Then, 1.5 mL of a solution prepared from 45 mg of sodium borohydride and 50mM NaOH was accurately added, and it was reacted in a 60°C oil bath pot for 3h. After the reaction was completed, glacial acetic acid was slowly added until no bubbles were generated to terminate the reaction. Then, a rotary evaporator was used to spin dry the reaction liquid at 45°C. In order to completely remove borate ions (BO 3- ), 5% acetic acid-methanol solution was added twice, each 2 mL, and spin dried respectively. Finally, chromatographic methanol was added to spin dry to further remove residual acid. After this series of treatments, a white powder sample was finally obtained. The sample was added with 2 mL of pyridine and 2 mL of acetic anhydride, the reaction environment was ensured to be closed, and it was reacted in a 95°C oil bath pot for 1h. After the reaction was completed, the reaction container was taken out and naturally cooled to room temperature. Then, the liquid in the bottle was transferred to a clean glass test tube, 3 mL of dichloromethane and 3 mL of ultrapure water were added, and extraction was performed by blowing. Then, the test tube was placed in a centrifuge and centrifuged at a speed of 4000 rpm for 10 minutes. After centrifugation, the supernatant was discarded, and the same amount of ultrapure water was added for two times of extraction. The lower organic layer was collected and blown dry with nitrogen. Then, 800 μL of dichloromethane was added to dissolve it completely. Finally, the redissolved sample was filtered through a 0.22 μm filter to remove possible impurities. The filtered sample was transferred to a liquid phase vial for testing. Agilent 7890A-5975C GC-MS was used for detection, and the detection conditions were as follows: HP-5MS capillary column (0.25x250 μm, 30m); helium gas was used as the carrier gas at a flow rate of 1 mL / min, 1 μL was injected, the initial temperature was 150°C, and it was maintained for 2 min with a split ratio of 1 / 8; then it was increased to 250°C at a rate of 2.5°C / min and maintained for 20 min; the MS quadrupole was 150°C, the ion source was 230°C, and the solvent delay was 5 min.

[0070] The results show that the polysaccharide of the present application mainly contains four kinds of sugar residue groups: Glcp-(1→, →6)-Fruf-(2→, →1)-Fruf-(2→, →1,6)-Fruf-(2→, and the molar percentages are 8.25:60.96:28.53:2.25, respectively. The ratio of the terminal residue (Glcp-(1→) to the branch site (→1,6)-Fruf-(2→) is close, and the branching degree of the polysaccharide of the present application is calculated to be 10.508% (Table 1). Figure 5

[0071] Table 1 Connection mode of sugar residues of the polysaccharide of the present application

[0072]

[0073] Example 7

[0074] NMR detection of the polysaccharide of the present application

[0075] Accurately weigh 50.0 mg of the polysaccharide obtained in Example 1, and then add 1 mL of deuterium water (D2O) to help it dissolve. Heat to 50°C in a water bath to accelerate the dissolution process. Then, place the solution in a refrigerator at -80°C to freeze to dryness. Then, place the frozen sample in a freeze dryer and freeze-dry it several times until it is a dry powder. This process is repeated three times, and after the last freeze-drying, 500 μL of D2O is added. Then, the sample is heated and the supernatant is separated by centrifugation (5000 rpm, 10 min). The supernatant is transferred to a NMR tube for subsequent detection. The uniform polysaccharide sample of the lophatherum is detected by 600M NMR spectrometer for one-dimensional nuclear magnetic spectrum (1H-NMR, 13C-NMR, DEPT spectrum) and two-dimensional nuclear magnetic spectrum (HSQC, HMBC). During the detection process, the detection time is appropriately extended to ensure the accuracy of the results.

[0076] The results show that the polysaccharide of the present application has α-D-Glcp-(1→, →6)-β-Fruf-(2→, →1,2)-β-Fruf-(6→ and →1)-β-Fruf-(2→ sugar residues. Among them, the fructose and glucose are connected by (2→1) position, and the fructose is connected by (2→1) or (2→6) position (Table 2). Figures 6 to 10

[0077] Table 2 Composition of the polysaccharide of the present application 1 H NMR and 13 C NMR data

[0078]

[0079] ​​Based on the nuclear magnetic resonance results, combined with the monosaccharide composition, infrared and ultraviolet spectra, and methylation results, the structure of the obtained polysaccharide is inferred as shown in Formula I:

[0080]

[0081] Where: n1 is 8.25, n2+n6 is 28.53, n3 is 2.25, and n4+n5 is 60.96.

[0082] Example 8

[0083] Effects of the polysaccharide of this invention on imiquimod-induced psoriasis in mice

[0084] The homogeneous polysaccharide of Platycodon grandiflorum obtained in Example 1 was compared with other homogeneous polysaccharides of inulin obtained from other medicinal materials in the following experiments:

[0085] Twelve male BALB / c mice were numbered, weighed, and their weight recorded. They were randomly divided into four groups based on body weight: a normal control group, a model control group, the polysaccharide group of this invention, and other inulin homogeneous polysaccharide groups, with three mice in each group. Treatment for each group was as follows: The animals were shaved on their backs the day before drug application, creating an exposed skin area of ​​approximately 2cm × 3cm. Except for the normal control group, the mice in the other groups received 50mg of 5% imiquimod cream every morning, and in the afternoon, the dosage of the polysaccharide of this invention and the other inulin homogeneous polysaccharides in each group was 100mg·kg⁻¹. -1· d -1 The normal control group and the model control group were given an equal volume of purified water once a day for 6 consecutive days. On the 7th day, photos were taken, and the severity of skin lesions (psoriasis area and severity index, PASI) was scored. Erythema, scales, and thickening were each scored from 0 to 4 (0, none; 1, mild; 2, moderate; 3, severe; 4, very severe). The three scores were accumulated to obtain the total score.

[0086] The results showed that the skin on the backs of mice in the normal control group was smooth, without scales, wrinkles, or erythema. After 6 days of imiquimod modeling, the skin on the backs of mice significantly thickened, with scales covering the entire hairless area, and erythema faintly visible beneath the scales. The wrinkles appeared as raised strips. The total PASI score was significantly higher than that of the normal control group (P < 0.01), indicating successful modeling. Compared with the model control group, the polysaccharide group of this invention showed a significant decrease in the scale infiltration erythema score (P < 0.01), while other inulin homogenized polysaccharide groups showed no significant difference (see...). Figure 11 (Table 3). This illustrates that not all inulin has anti-psoriasis effects; the specific structure of the polysaccharides in this invention determines their efficacy.

[0087] Table 3 Effect of polysaccharide of the application on imiquimod-induced psoriasis mouse model (n=3)

[0088]

[0089] Compared with the normal control group, ΔΔ P<0.01, compared with the model control group, ** P<0.01

[0090] The preparation method of other inulin homopolysaccharide in this embodiment is as follows:

[0091] (1) The sliced atractylodes rhizome was crushed, passed through a No. 5 sieve, and heated to reflux with 5 times the amount of 70% ethanol for 2 h to remove fat. The step was repeated once, and the dried atractylodes rhizome was obtained. After the atractylodes rhizome was defatted, 400 mL of pure water was added at a solid-liquid ratio of 1:20 (m / v), and the mixture was soaked for 2 h. Then, the mixture was placed in an ultrasonic sample processing machine under the following conditions: ultrasonic frequency 25 kHz, ultrasonic power 300 W, ultrasonic temperature 60°C, and ultrasonic time 30 min. The polysaccharide was extracted by ultrasonic extraction, and the hot filtrate was filtered through 8 layers of gauze. The extraction was repeated twice, and the combined filtrate was concentrated to a crude drug amount of 0.5 g / mL in a rotary evaporator (60°C). Then, 95% ethanol was slowly added to the concentrated solution until the final ethanol volume fraction was 86% (v / v). The mixture was placed at 4°C overnight, and the precipitate was repeatedly washed with anhydrous ethanol and centrifuged twice. The supernatant was clear, and the precipitate was redissolved in pure water. The solution was poured into a 3000 Da dialysis bag (activated at 100°C for 10 min and washed with pure water), and dialyzed for 72 h (3-4 h per water change). Finally, the solution was freeze-dried to obtain the total polysaccharide sample of atractylodes rhizome.

[0092] (2) 1.5 g of the total polysaccharide of atractylodes rhizome was weighed and dissolved in 100 mL of pure water. The mixture was heated in a 60°C water bath for 30 min, centrifuged at 4000 rpm for 10 min, and the supernatant was collected. The supernatant was separated by HPLC using 0 M, 0.1 M, 0.2 M, 0.4 M, and 0.8 M NaCl as the mobile phase at a flow rate of 3 mL / min. The eluate was collected in separate tubes using a programmable automatic fraction collector, with 10 mL per tube and 100-300 tubes per fraction. The sugar content of the eluate was determined by the phenol-sulfuric acid method and the enzyme label instrument, and the absorbance was used to draw the elution curve. The eluate containing polysaccharide was collected according to the elution curve, concentrated to 50 mL by rotary evaporation (60°C), and dialyzed against pure water several times a day. After 72 h of dialysis, the solution in the dialysis bag was freeze-dried. The portion eluted by 0.1 M NaCl solution was the other inulin homopolysaccharide. The structure of the other inulin homopolysaccharide was determined by ultraviolet, infrared, methylation experiment, monosaccharide composition, and nuclear magnetic resonance, and the structure formula is shown as formula II:

[0093]

[0094] Wherein: n1 is 2.32, n2 is 28.75, n3 is 2.25, n4 is 50.46, n5 is 4.78, n6 is 4.89, n7 is 6.55.

[0095] Example 9

[0096] A tablet for treating psoriasis is prepared by using the polysaccharide of Example 1 200g, starch 300g, magnesium stearate 5g and sodium carboxymethyl cellulose 10g in a manner known in the art.

[0097] Example 10

[0098] A capsule for treating psoriasis is prepared by weighing the polysaccharide of Example 1 200g, mixing, adding magnesium stearate 5g, mixing, and filling into a capsule.

[0099] Example 11

[0100] A granule for treating psoriasis is prepared as follows:

[0101] 1. β-cyclodextrin is dissolved in water by heating and stirring; 2. the polysaccharide of Example 1 is dissolved in water, and then slowly added dropwise into the β-cyclodextrin aqueous solution, the mass ratio of β-cyclodextrin to polysaccharide being 6-8:1, and stirring; 3. after cooling, the crystals are filtered, washed, dried, and pulverized to pass through a 40-mesh screen (Chinese Pharmacopoeia screen) to obtain the product.

[0102] Example 12

[0103] A pill for treating psoriasis is prepared by grinding the polysaccharide of the present application, passing through a 100-mesh screen, spreading the powder in a coating pan, and preparing into a water pill in a manner known in the art.

[0104] Example 13

[0105] An oral solution for treating psoriasis is prepared by dissolving the polysaccharide of Example 1 in water at a mass ratio of 1:10, adding 0.03% nipagin acetate as a preservative, 0.02% ascorbic acid as an antioxidant, and 0.02% of a flavoring agent; the prepared polysaccharide solution is filtered, the filtrate is clear for 8 hours to obtain a clear solution; the clear polysaccharide solution is divided into 10-mL glass bottles, and the divided polysaccharide solution is sterilized at high temperature, at 121°C / 0.125 MPa for 20 minutes to obtain the product.

[0106] The above examples are the preferred embodiments of the present application, but the embodiments of the present application are not limited by the above examples, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application are equivalent replacement methods and are included in the protection scope of the present application.

Claims

1. A polysaccharide derived from bellflower inulin, characterized in that: It contains fructose and glucose in a molar ratio of 95.6:4.4; the weight-average molecular weight of the inulin polysaccharide is 2-10 kDa. The aforementioned inulin polysaccharide is based on fructose and has the structure shown in Formula I: In Formula I, n1 is 1 to 15, (n2+n6) is 20 to 40, n3 is 1 to 10, and (n4+n5) is 30 to 90.

2. The inulin polysaccharide according to claim 1, characterized in that: The weight-average molecular weight of the inulin polysaccharide is 3-5 kDa.

3. The inulin polysaccharide according to claim 1, characterized in that: The weight-average molecular weight of the inulin polysaccharide is 3.8 kDa.

4. The inulin polysaccharide according to claim 1, characterized in that: In Formula I, n1 is 6 to 10, (n2+n6) is 25 to 30, n3 is 2 to 4, and (n4+n5) is 55 to 65.

5. The inulin polysaccharide according to claim 1, characterized in that: In Equation I, n1 is 8.25, (n2+n6) is 28.53, n3 is 2.25, and (n4+n5) is 60.

96.

6. The method for preparing the polysaccharide from bellflower inulin according to any one of claims 1 to 5, characterized in that: Includes the following steps: (1) Take the medicinal slices of Platycodon grandiflorus, defatt them, then extract them with alkaline water, concentrate the extract, precipitate them with ethanol solution, redissolve the precipitate with water, remove the protein using the Sevage method, freeze dry them to obtain the total polysaccharides of Platycodon grandiflorus. (2) The total polysaccharide of Platycodon grandiflorus was redissolved, centrifuged, and the supernatant was added to the chromatography column. The eluent was collected, concentrated, and dialyzed to obtain the Platycodon grandiflorus inulin polysaccharide.

7. The preparation method according to claim 6, characterized in that: The step of collecting the eluent in step (2) is as follows: During the elution process, the equilibrium flow rate was set to 2.5 min / 10 mL, and water, 0.1 mol / L, 0.2 mol / L, 0.4 mol / L, 0.8 mol / L, and 1 mol / L NaCl were used as mobile phases for elution in sequence, and the eluent of 0.1 M NaCl solution was collected.

8. The use of the platycodon inulin polysaccharide according to any one of claims 1 to 5 in the preparation of a drug for treating psoriasis.

9. The application according to claim 8, characterized in that: The aforementioned psoriasis treatment drug comprises a therapeutically effective amount of the said inulin polysaccharide and a pharmaceutically acceptable carrier.

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