Platycodon grandiflorum inulin polysaccharide, preparation method thereof and application of platycodon grandiflorum inulin polysaccharide in preparation of medicine for treating psoriasis

Abstract

The invention discloses platycodon grandiflorum inulin polysaccharide as well as a preparation method and application thereof in preparation of a medicine for treating psoriasis, the structure of the platycodon grandiflorum inulin polysaccharide is as shown in formula I, the platycodon grandiflorum inulin polysaccharide contains fructose and glucose, and the weight-average molecular weight of the platycodon grandiflorum inulin polysaccharide is 2-10KD. The platycodon grandiflorum inulin polysaccharide disclosed by the invention is remarkable in psoriasis resisting effect, safe, non-toxic and good in biocompatibility, and is expected to be developed into a novel treatment medicine for treating psoriasis. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of natural medicines, and in particular relates to platycodon inulin polysaccharide and a preparation method thereof and application thereof in preparing a medicine for treating psoriasis. Background Art

[0002] Psoriasis is a common chronic immune-mediated proliferative skin disease, and its main skin pathological features include epidermal hyperplasia, obvious proliferation of keratinocytes, increased and expanded angiogenesis, and obvious inflammatory infiltration. There are many patients with psoriasis, and many new cases are reported every year. The etiology of psoriasis is complex and may be caused by multiple factors such as genetic factors, environmental factors, and autoimmune factors. However, its pathogenesis is still unclear, so its condition is stubborn and difficult to cure, and it is prone to recurrence. At the same time, it also increases the possibility and incidence of metabolic syndrome and cardiovascular disease, bringing heavy life burden and mental stress to patients.

[0003] At present, the commonly used therapeutic drugs in clinical practice mainly include topical drugs, traditional therapeutic drugs, biological agents, small molecule targeted drugs, etc. Topical drugs include tamoxifen, vitamin D3 derivatives, anthralin, tretinoin cream, glucocorticoids, etc.; traditional therapeutic drugs include methotrexate, cyclosporine, acitretin, etc.; biological agents include TNF-α inhibitors (etanercept, adalimumab), IL-12 / 23 inhibitors (ustekinumab), IL-23 inhibitors (gusekizumab), etc.; small molecule targeted drugs include PDE4 inhibitors (apmilast), JAK1-3 inhibitors (tofacitinib, upadacitinib), TYK2 inhibitors (deucravacitinib), etc.

[0004] Topical medications are easy for patients to accept and have high compliance, but they are ineffective for moderate to severe patients; traditional treatment drugs are effective and easy to use, but they can cause adverse reactions such as liver and kidney toxicity, hypertension and hyperlipidemia, and the efficacy of first-line drug methotrexate is only 30%-45%; biological agents have better efficacy, but the patient's health status needs to be fully evaluated before treatment, and due to the short clinical application time, its long-term efficacy and safety remain to be observed; small molecule targeted drugs have large differences in efficacy and safety in the treatment of psoriasis, and there is a risk of serious adverse reactions. Therefore, finding new safe and effective treatment drugs is an urgent need for the treatment of psoriasis. Summary of the invention

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

[0006] The purpose of the present invention is achieved through the following technical solutions:

[0007] A platycodon inulin polysaccharide, containing fructose and glucose, the molar concentration ratio of which is 95.6:4.4;

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

[0009] A platycodon inulin polysaccharide, with fructose as the main chain and a structure as shown in Formula I:

[0010]

[0011] The numbers in the main chain and side chain represent the attachment sites, and the arrows represent the attachment directions.

[0012] The weight average molecular weight of the Platycodon inulin polysaccharide is 2 to 10 KD;

[0013] 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;

[0014] Preferably, the weight average molecular weight of the Platycodon inulin polysaccharide is 3 to 5 KD;

[0015] 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;

[0016] Particularly preferably, the weight average molecular weight of the Platycodon inulin polysaccharide 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 platycodon inulin polysaccharide comprises the following steps:

[0019] (1) taking a decoction piece of Platycodon grandiflorum, defatting it, then extracting it with alkaline water, concentrating the extract, precipitating it with an ethanol solution, re-dissolving the precipitate with water, deproteinizing it with a Sevage method, and freeze-drying it to obtain Platycodon grandiflorum total polysaccharide;

[0020] (2) re-dissolving the total polysaccharide of Platycodon grandiflorum, centrifuging, taking the supernatant and adding it to a chromatography column, collecting the eluate, concentrating and dialyzing to obtain the Platycodon grandiflorum inulin polysaccharide of the present invention;

[0021] Step (1) defatting uses 70% ethanol solution.

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

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

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

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

[0026] 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 in sequence. The eluate of 0.1 M NaCl solution was collected.

[0027] The dialysis in step (2) is performed for 3 days in a dialysis bag filled with 3000Da, during which the dialysis water is replaced every day.

[0028] Animal experiments have shown that the Platycodon inulin polysaccharide of the present invention has a significant anti-psoriasis effect and can be used to prepare a drug for treating psoriasis.

[0029] The drug for treating psoriasis comprises a therapeutically effective amount of the platycodon inulin polysaccharide and a pharmaceutically acceptable carrier, wherein the carrier includes pharmaceutically commonly used excipients such as water, starch, lactose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, etc., preservatives such as sodium benzoate and ethyl paraben, glidants such as magnesium stearate, disintegrants such as sodium carboxymethyl cellulose, etc.

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

[0031] Compared with the prior art, the present invention has the following advantages and effects:

[0032] The platycodon inulin polysaccharide of the present invention has significant anti-psoriasis effect, is safe and non-toxic, and has good biocompatibility, and is expected to be developed into a new therapeutic drug for treating psoriasis. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0034] Figure 2 This is the HPGPC diagram of the polysaccharide of the present invention.

[0035] Figure 3 This is a diagram showing the monosaccharide composition of the polysaccharide of the present invention.

[0036] Figure 4is the infrared spectrum of the polysaccharide of the present invention.

[0037] Figure 5 This is the total ion current diagram of the methylated sugar alcohol acetyl ester derivative of the polysaccharide of the present invention.

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

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

[0040] Figure 8 It is the DEPT spectrum of the polysaccharide of the present invention.

[0041] Fig. 9 The HSQC-NMR spectrum of the polysaccharide of the present invention.

[0042] Fig.10 The HMBC-NMR spectrum of the polysaccharide of the present invention.

[0043] Fig.11 This figure shows the effect of the polysaccharide of the present invention on the imiquimod-induced psoriasis model in BALB / c mice. DETAILED DESCRIPTION

[0044] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0045] Example 1

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

[0047] (1) Grind the slices of Platycodon grandiflorum, pass through a No. 6 sieve, heat and cycle with 70% ethanol to remove fat, and dry to obtain defatted medicinal materials. Take the defatted medicinal materials, add 25 times the pH value of 12 ammonia solution, heat and cycle to extract for 2 hours, after hot filtration, extract the residue again, combine the filtrate, adjust the pH value to 7 with acetic acid aqueous solution, concentrate under reduced pressure to a crude drug concentration of 0.2g / mL, precipitate with 85% ethanol, centrifuge, wash with anhydrous ethanol several times, and redissolve with ultrapure water. Transfer to a 3000Da dialysis bag for 72 hours, replace pure water every 4 hours, freeze-dry, and weigh to obtain Platycodon grandiflorum total polysaccharides.

[0048] (2) Take 2 g of dried Platycodon grandiflorum total polysaccharide and add 100 mL of pure water to fully dissolve it. Then, centrifuge at 3000 rpm for 15 min, take the supernatant and load it onto 200 tubes of DEAE-52 cellulose anion exchange chromatography columns, 10 mL per tube, and use the phenol-sulfuric acid method to determine the polysaccharide content, thereby drawing an elution curve ( 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 pure water was changed more than 6 times a day. Finally, the solution in the dialysis bag was freeze-dried, and the part eluted with 0.1 M NaCl solution was the polysaccharide of the present invention.

[0049] Example 2

[0050] Determination of sugar content and protein content of the polysaccharide of the present invention

[0051] 5.0 mg of fructose standard was accurately weighed, ultrapure water was added to prepare a 1 mg / mL fructose standard solution, and the solution was continuously diluted in half to obtain a series of fructose standard solutions of 500, 250, 125, 62.5, and 31.25 μg / mL. Similarly, 5.0 mg of the polysaccharide obtained in Example 1 was accurately weighed, dissolved in ultrapure water to prepare a 1 mg / mL sample solution, and diluted to 500 μg / ml for standby use. The polysaccharide content was detected by the 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, and then 500 μL of concentrated sulfuric acid solution was added and shaken, and after cooling, 150 μL of the reaction solution was drawn into a 96-well plate, and the absorbance was detected at 490 nm with an enzyme marker. The fructose standard curve was drawn with the standard concentration as the horizontal coordinate and the absorbance as the vertical coordinate, and a linear equation was obtained to calculate the polysaccharide content. The polysaccharide protein content of the present invention was measured by the BCA kit method.

[0052] The results showed that the sugar content of the polysaccharide of the present invention was 96.58%, and the protein content was undetectable.

[0053] Example 3

[0054] Example 1 Determination of Molecular Weight of the Polysaccharide Obtained

[0055] Take 3 mg of 3690, 2370, 1190, 680, 450, 35 and 4Kd dextran standards respectively, prepare 1.0 mg / mL reference solution with ultrapure water, pass through 0.45 μm water filter membrane, and detect by Agilent 1260 high performance liquid chromatograph (equipped with differential detector). Draw the standard curve with the average molecular weight logarithm log (Mw) of the standard as the horizontal axis and the retention time as the vertical axis. Chromatographic analysis conditions: TSKG-5000PWXL chromatographic column, water as mobile phase, column temperature of 40°C, flow rate set to 0.4 mL / min, differential refractometer temperature also set to 40°C, injection volume set to 10 μL. Take 3 mg of sample accurately, test according to the above procedure, record the retention time, and import it into the standard equation to calculate the molecular weight distribution of polysaccharide.

[0056] The results showed that the polysaccharide of the present invention was a single symmetrical peak with a molecular weight of 3800Da ( Figure 2).

[0057] Example 4

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

[0059] Preparation of test sample: 5.0 mg of the polysaccharide sample obtained in Example 1 was accurately weighed and placed in a 50 mL round-bottom flask, followed by the addition of 4 mL of a 0.5 M trifluoroacetic acid solution. The flask was placed in an oil bath and hydrolyzed at 80°C with stirring for 40 minutes. After the hydrolysis was completed, the reaction solution was transferred to a rotary evaporator and dried. Subsequently, 4 mL of chromatographic methanol was added and the solution was dried under reduced pressure again until there was no sour taste, so as to completely remove the residual trifluoroacetic acid.

[0060] Preparation of mixed standard: Accurately weigh 5 mg of each of Glc, fructose (Fru), Man, Rha, Gala, Gal, GclA, and Ara standards. Then add 5 mL of ultrapure water and fully dissolve to prepare a 30 mg / mL mixed standard solution. To ensure the accuracy of the experiment, the mixed solution needs to be further diluted to prepare a concentration series of 3000, 1500, 750, 375, 187.5, and 93.75 μg / mL in sequence for use.

[0061] Accurately pipette 50 μL of the polysaccharide solution obtained in Example 1 and the standard solution of different concentrations, and add 100 μL of methanol solution of p-aminobenzoic acid respectively. Subsequently, place the reaction tube in a metal bath at 70°C and heat the reaction for 40 minutes. Take out the test tube and cool it to room temperature. Then, add ultrapure water to dilute to 2.5mL, ensure that the sample is fully mixed, filter with a 0.45μm microporous filter membrane, and transfer the treated sample to a liquid phase vial for testing. The liquid chromatography conditions are as follows: the temperature of the chromatographic column is 30°C, the mobile phase flow rate is set to 0.8mL / min, and the sample injection volume is 10μL. The wavelength of the ultraviolet detector (DAD) is set to 303nm, and the excitation wavelength and emission wavelength of the fluorescence detector (FLD) are 313nm and 358nm, respectively. The mobile phase B phase is 0.1M phosphate buffer (pH=2.54) containing tetrabutylammonium sulfate, and the A phase is a mixture of phase B and chromatographic methanol in a volume ratio of 1:1. The liquid mobile phase will be 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 showed that the polysaccharide of the present invention mainly contained fructose and glucose, and the molar concentration ratio was 95.6:4.4 ( Figure 3 ).

[0063] Example 5

[0064] Infrared spectroscopic determination of polysaccharides of the present invention

[0065] Take 3 mg of polysaccharide obtained in Example 1 and precipitate it at 4000-400 cm -1 Infrared measurements were performed using a Fourier transform infrared spectrometer.

[0066] The results showed that the polysaccharide of the present invention has a peak at 3268 cm -1 The broad and deep peak at 2930 cm indicates the presence of intermolecular OH vibration, while the -1 and 2882cm -1 The peak at 1120cm -1 and 1032cm -1 The peaks at 933 cm correspond to the OH angle vibration and the ether bond COO vibration. -1 、870cm -1 and 820cm -1 The peak of Figure 4 ).

[0067] Example 6

[0068] Polysaccharide methylation assay of the present invention

[0069] 20.0 mg of the polysaccharide obtained in Example 1 was placed in a 5 mL centrifuge tube and dried at 80 ° C for 10 min. Then, 4 mL of anhydrous dimethyl sulfoxide (DMSO) was added, the air was replaced with nitrogen and the airtight ultrasonic treatment was performed for 2 h, then 2 mL of anhydrous DMSO-NaOH suspension was added, the air in the bottle was replaced with nitrogen again, and the ultrasonic reaction was performed at a temperature of 18 to 20 ° C for 1 h. Iodomethane was slowly added under light-proof conditions, added three times, 500 μL each time, and a total of 1.5 mL of iodomethane was added, and then the light-proof ultrasonic reaction was continued at 18 to 20 ° C for 40 minutes. After the reaction was completed, 1 mL of ultrapure water was added to terminate the reaction, and the solution was dialyzed overnight in a 3000 Da dialysis bag. The next day, the liquid in the dialysis bag was poured into a separatory funnel, and an equal amount of dichloromethane was added for extraction. After standing for 10 minutes, the upper aqueous layer was discarded, and an equal amount of ultrapure water was added to continue extraction. Perform three extractions in total, pour the lower organic layer into a clean glass centrifuge tube and dry to obtain a powdered sample. Infrared detection 3700~3200cm -1Whether the hydrogen hydroxyl functional groups within the range react completely. Take the completely dried methylated sample, accurately add 2mL of 88% formic acid, and after fully dissolving, transfer it to a dry and clean round-bottom flask. Subsequently, nitrogen replacement is performed to ensure that the reaction environment is an inert atmosphere, and the reaction is sealed in an oil bath at 100°C for 4h. After the reaction is completed, the flask is taken out and naturally cooled to room temperature. Use a rotary evaporator to spin dry the reaction solution at 45°C, then add 3mL of chromatographic methanol, and continue to evaporate until the sour taste is completely removed to obtain the treated sample. Take the sample obtained in the previous step, add 3mL of ultrapure water to dissolve, take 0.5mL for subsequent reactions, add 1.5mL of ultrapure water, 2mL of 4M trifluoroacetic acid solution (the final reaction solution is 2M trifluoroacetic acid), and react at 80°C for 4h. After the end, let it cool, decompress the rotary evaporator and spin dry (45°C), add chromatographic methanol and spin dry to remove acid. Take the sample obtained in the previous step and add 1.5mL of 50mM NaOH solution to dissolve. Then, 1.5 mL of a solution prepared from 45 mg of sodium borohydride and 50 mM NaOH was accurately added and reacted in an oil bath at 60 °C for 3 h. After the reaction was completed, glacial acetic acid was slowly added until no bubbles were generated to terminate the reaction. Subsequently, the reaction solution was dried using a rotary evaporator at 45 °C. In order to completely remove the borate ions (BO 3- ), then add 2mL of 5% acetic acid-methanol solution twice, and spin dry respectively. Finally, add chromatographic methanol for rotary evaporation to further remove the residual acid. After this series of treatments, a white powdery sample is finally obtained. Add 2mL pyridine and 2mL acetic anhydride to the sample, ensure that the reaction environment is closed, and react in an oil bath at 95°C for 1h. After the reaction is completed, take out the reaction vessel and cool it naturally to room temperature. Then, transfer the liquid in the bottle to a clean glass test tube, add 3mL dichloromethane and 3mL ultrapure water, and extract by blowing. Subsequently, put the test tube into a centrifuge and centrifuge at 4000rpm for 10 minutes. After centrifugation, discard the supernatant and add an equal amount of ultrapure water for two extractions. Collect the lower organic layer and blow it dry with nitrogen. After that, add 800μL dichloromethane for redissolution to ensure that the sample is completely dissolved. Finally, filter the redissolved sample through a 0.22μm filter head to remove possible impurities. The filtered sample was transferred to a liquid phase vial for testing. The Agilent 7890A-5975C GC-MS was used for detection. The detection conditions were: HP-5MS capillary column (0.25×250μm, 30m); helium as carrier gas at a flow rate of 1mL / min, 1μL injection, initial temperature of 150℃, maintained for 2min, split ratio: 1 / 8; then increased to 250℃ at 2.5℃ / min and maintained for 20min; MS quadrupole 150℃, ion source 230℃, solvent delay 5min.

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

[0071] Table 1 Sugar residue connection mode of the polysaccharide of the present invention

[0072]

[0073] Example 7

[0074] Polysaccharide nuclear magnetic resonance detection of the present invention

[0075] Accurately weigh 50.0 mg of the polysaccharide obtained in Example 1, and then add 1 mL of deuterated water (D2O) to help it dissolve. Heat to 50 ° C in a water bath to accelerate the dissolution process. Subsequently, the solution is placed in a -80 ° C refrigerator and frozen to a dry state. Then, the frozen sample is placed in a freeze dryer and subjected to multiple freeze-drying processes until it is a dry powder. This process requires three freeze-thaw cycles, and after the last freeze-drying, 500 μL of D2O is added. Subsequently, the sample is heated and the supernatant is separated by centrifugation (5000 rpm, 10 min). The supernatant is transferred to a nuclear magnetic tube for subsequent detection. The one-dimensional nuclear magnetic spectrum (1H-NMR, 13C-NMR, DEPT spectrum) and two-dimensional nuclear magnetic spectrum (HSQC, HMBC) of the Platycodon grandiflorum uniform polysaccharide sample are detected using a 600M nuclear magnetic resonance spectrometer. 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 invention is composed of α-D-Glcp-(1→, →6)-β-Fruf-(2→, →1,2)-β-Fruf-(6→ and →1)-β-Fruf-(2→ sugar residues. Among them, fructose and glucose are connected at the (2→1) position, and fructose is connected at the (2→1) or (2→6) position. Figures 6 to 10 , Table 2).

[0077] Table 2 Polysaccharides of the present invention 1 H NMR and 13 C NMR data

[0078]

[0079] According to the NMR results, combined with the monosaccharide composition, infrared, ultraviolet spectra and methylation results, the structure of the obtained polysaccharide is inferred to be as shown in Formula I:

[0080]

[0081] Among them: 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 the present invention on imiquimod-induced psoriasis mice

[0084] The Platycodon inulin uniform polysaccharide obtained in Example 1 was used together with other inulin uniform polysaccharides isolated from other medicinal materials to conduct the following experiment:

[0085] Twelve male BALB / c mice were numbered, weighed and recorded. They were randomly divided into 4 groups according to their body weight, namely normal control group, model control group, polysaccharide group of the present invention and other inulin uniform polysaccharide groups, with 3 mice in each group. The treatment of each group was as follows: the hair on the back of the animals was shaved one day before the application of the drug to form a skin exposure area of ​​about 2cm×3cm. Except for the normal control group, the mice in the remaining groups were smeared with 5% imiquimod cream 50mg in the morning, and the drug dose of the polysaccharide of the present invention and other inulin uniform polysaccharides in each group was 100mg·kg in the afternoon. -1· d -1 The normal control group and the model control group were given an equal volume of pure 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. The erythema, scales and thickness were each 0-4 (0, none; 1, mild; 2, moderate; 3, severe; 4, very severe), and the three scores were accumulated to obtain the total score.

[0086] The results showed that the back skin of mice in the normal control group was smooth, without scales, wrinkles, erythema, etc. After 6 days of imiquimod modeling, the back skin was significantly thickened, scales covered the entire depilatory area, erythema covered by scales was faintly visible, and wrinkles were strip-shaped and raised. The PASI total score was significantly higher than that of the normal control group (P < 0.01), indicating that the modeling was successful. Compared with the model control group, the scale infiltration and erythema score of the polysaccharide group of the present invention was significantly reduced (P < 0.01), while there was no significant difference in other inulin uniform polysaccharide groups (see Fig.11 , Table 3). This indicates that not all inulins have anti-psoriatic effects, and the specific structure of the polysaccharide of the present invention determines its efficacy.

[0087] Table 3 Effects of the polysaccharide of the present invention on the psoriasis mouse model induced by imiquimod (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 uniform polysaccharides in this embodiment is as follows:

[0091] (1) Crush the sliced ​​Atractylodes lancea, pass through a No. 5 sieve, and use 5 times the amount of 70% ethanol to heat and reflux for 2 hours to remove fat. Repeat this step once and dry to obtain defatted medicinal materials. Take the defatted Atractylodes lancea powder, add 400 mL of pure water at a drug-liquid ratio of 1:20 (m / v), soak for 2 hours, put it in an ultrasonic sample processor, and ultrasonically extract Atractylodes lancea polysaccharides under the conditions of ultrasonic frequency of 25 kHz, ultrasonic power of 300 W, ultrasonic temperature of 60°C, and ultrasonic time of 30 minutes. Filter while hot through 8 layers of gauze, extract twice, and combine the filtrate. The filtrate was concentrated to a crude drug amount of 0.5 g / ml in a rotary evaporator (60°C), 95% ethanol was added to the concentrate (slowly and quickly stirred) to a final ethanol volume fraction of 86% (v / v), and the mixture was allowed to stand at 4°C for alcohol precipitation overnight. The mixture was centrifuged and the precipitate was repeatedly washed with anhydrous ethanol and centrifuged twice. The supernatant became clear, and pure water was added for reconstitution. The mixture was poured into a 3000Da dialysis bag (activated at 100°C for 10 min, and washed with pure water) and dialyzed for 72 h (3-4 h / water change), and freeze-dried to obtain a total polysaccharide sample of Atractylodes lancea.

[0092] (2) Weigh 1.5 g of total polysaccharides from Atractylodes lancea, dissolve in 100 mL of pure water, place in a 60°C water bath for 30 min, centrifuge at 4000 rpm for 10 min, take the supernatant and load it, use 0 M, 0.1 M, 0.2 M, 0.4 M, and 0.8 M NaCl as the mobile phase, balance the flow rate with a digital constant flow pump at 3 mL / min, and use a programmable automatic fraction collector to collect the eluate in separate tubes, 10 mL / tube, collect 100 to 300 tubes for each part, and draw 200 μL of the eluate from the tube for phenol-sulfuric acid method and microplate reader to detect the sugar content, and draw the elution curve with absorbance. Collect the eluate containing polysaccharides according to the elution curve, concentrate to 50 mL by rotary evaporation (60°C), dialyze with pure water, change the pure water several times a day, and after dialysis for 72 hours, finally freeze-dry the solution in the dialysis bag, of which the part eluted with 0.1 M NaCl solution is other inulin uniform polysaccharides. Its structural formula is shown in Formula II as determined by various methods including UV, IR, methylation experiments, monosaccharide composition, and NMR:

[0093]

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

[0095] Example 9

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

[0097] Example 10

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

[0099] Embodiment 11

[0100] A granule for treating psoriasis, the preparation method is as follows:

[0101] ① β-cyclodextrin is dissolved in water, heated and stirred to dissolve; ② After the polysaccharide of Example 1 is dissolved in water, it is slowly dripped into the β-cyclodextrin aqueous solution, the mass ratio of β-cyclodextrin to polysaccharide is 6-8:1, and stirred; ③ After cooling, filter to obtain crystals, wash, dry, crush, and pass through a 40-mesh sieve (Chinese Pharmacopoeia sieve).

[0102] Example 12

[0103] A pill for treating psoriasis is prepared by grinding the polysaccharide of the present invention, passing the powder through a 100-mesh sieve, spreading the powder in a coating pan, and using a method known in the art to prepare water pills.

[0104] Embodiment 13

[0105] An oral liquid for treating psoriasis is prepared by dissolving the polysaccharide of Example 1 in water at a mass ratio of 1:10, adding 0.03% of paraben acetate as a preservative, 0.02% of ascorbic acid as an antioxidant, and 0.02% of edible flavor; filtering the prepared polysaccharide solution, and clarifying the filtrate for 8 hours to obtain a clarified solution; packaging the clarified polysaccharide solution into 10 mL glass bottles, and sterilizing the packaged polysaccharide solution at high temperature, using 121° C. / 0.125 MPa for 20 minutes to obtain an oral liquid.

[0106] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A Platycodon inulin polysaccharide, characterized in that: It contains fructose and glucose, and the molar concentration ratio of the fructose and glucose is 95.6:4.4; the weight average molecular weight of the platycodon inulin polysaccharide is 2-10KD.

2. A Platycodon inulin polysaccharide, characterized in that: With fructose as the main chain, the structure is shown in Formula I: The weight average molecular weight of the Platycodon inulin polysaccharide is 2 to 10 KD; 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.

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

4. The Platycodon inulin polysaccharide according to claim 1 or 2, characterized in that: The weight average molecular weight of the Platycodon inulin polysaccharide is 3.8KD.

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

6. The Platycodon inulin polysaccharide according to claim 2, characterized in that: In the formula I, n1 is 8.25, (n2+n6) is 28.53, n3 is 2.25, and (n4+n5) is 60.

96.

7. The method for preparing Platycodon inulin polysaccharide according to any one of claims 1 to 6, characterized in that The steps include: (1) taking a decoction piece of Platycodon grandiflorum, defatting it, then extracting it with alkaline water, concentrating the extract, precipitating it with an ethanol solution, re-dissolving the precipitate with water, deproteinizing it with a Sevage method, and freeze-drying it to obtain Platycodon grandiflorum total polysaccharide; (2) The total polysaccharide of Platycodon grandiflorum is redissolved, centrifuged, the supernatant is added to a chromatography column, the eluate is collected, concentrated, and dialyzed to obtain the Platycodon grandiflorum inulin polysaccharide of the present invention.

8. The preparation method according to claim 7, characterized in that: The step of collecting the eluate in step (2) is: 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 in sequence, and the eluate of 0.1 M NaCl solution was collected.

9. Use of the Platycodon inulin polysaccharide according to any one of claims 1 to 6 in the preparation of a drug for treating psoriasis.

10. The use according to claim 9, characterized in that: The drug for treating psoriasis comprises a therapeutically effective amount of the platycodon inulin polysaccharide and a pharmaceutically acceptable carrier.

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