Application of sarcandra glabra polysaccharide in preparation of medicine for preventing and / or treating psoriasis and delaying relapse of psoriasis

Abstract

The invention discloses an application of sarcandra glabra polysaccharide in preparation of a medicine for preventing and / or treating psoriasis and delaying relapse of psoriasis. The glabrous sarcandra herb polysaccharide can relieve inflammation caused by psoriasis, reduce expression of memory T cells at skin lesions of a psoriasis recurrence model mouse and relieve psoriasis symptoms of the recurrence model mouse. The invention finds that the glabrous sarcandra herb polysaccharide can obviously improve the skin injury degree of an imiquimod (imiquimod, IMQ) interval induced psoriasis recurrence model mouse, reduce the skin erythema, thickening and scale of the IMQ interval induced psoriasis recurrence model mouse, and reduce the expression of memory T cells at the skin injury part of the psoriasis recurrence model mouse, so that the glabrous sarcandra herb polysaccharide can be used for treating the psoriasis. The glabrous sarcandra herb polysaccharide has great application prospects in preparation of the medicine for preventing / treating the psoriasis, and especially can relieve recurrence symptoms of the psoriasis.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of traditional Chinese medicine, and particularly relates to application of Sarcandra glabra polysaccharide in preparation of a medicine for preventing and / or treating psoriasis and delaying recurrence of psoriasis. BACKGROUND

[0002] Psoriasis is a chronic inflammatory skin disease, and the total effective rate of traditional drugs is not more than 50%, and the disease is prone to recurrence after drug withdrawal.

[0003] Patients with mild psoriasis often use topical glucocorticoids, patients with moderate or severe psoriasis often use systemic therapy such as methotrexate and cyclosporine, and patients who do not respond to systemic therapy use biological agents, but no matter what kind of therapy, recurrence is a difficult problem that cannot be avoided.

[0004] Literature data shows that glucocorticoids relapse about 4 weeks after drug withdrawal (Zheng J. National Symposium on Integrative Dermatology and Venereology, Guiyang, Guizhou, China, 2018); the first-line drugs for treating psoriasis, such as methotrexate and cyclosporine, relapse about 4 weeks after drug withdrawal (Okubo, Y, J DERMATOL, 2010; 38(5): 465-72; Heydendael, VM, NEW ENGL J MED, 2003; 349(7): 658-65).

[0005] Compared with topical administration and traditional systemic administration, biological agents can achieve good clearance of skin lesions, but still cannot overcome the recurrence of psoriasis. The average recurrence time of psoriasis patients treated with TNF inhibitors (etanercept) etanercept is 7.3 weeks (Griffiths, CE, NEW ENGL J MED, 2010; 362(2): 118-28). IL-17RA antagonists (brodalumab) can cause recurrence in some patients 2 weeks after drug withdrawal, and the severity of the recurrence is higher than that of the initial recurrence, and more than that, recurrence occurs within a few days of drug withdrawal, and about half of the patients have recurrence within 50 days of drug withdrawal (MassonRegnault, M, J EUR ACAD DERMATOL, 2017; 31(9): 1491-1496). Biological agents targeting IL-23 show excellent durability, and the median time to recurrence of Risankizumab is 42 weeks (Blauvelt, A, JAMA DERMATOL, 2020; 156(6): 649-658); tildrakizumab delays recurrence for about 24 weeks and is not affected by dose (Kimball, AB, BRIT J DERMATOL, 2019; 182(6): 1359-1368. Warren, RB, J EUR ACAD DERMATOL, 2020; 35(4): 919-927). Therefore, psoriasis recurrence is the focus and difficulty of current psoriasis treatment, and reducing the symptoms of psoriasis recurrence is conducive to enhancing the effect of psoriasis treatment.

[0006] Sarcandra glabra (Thunb.) Nakai is a dry whole plant of grass coral Sarcandra glabra (Thunb.) Nakai of Hernandiaceae, also known as Jiegu Feng, Jiegu Cha, Jiegu Hua, Zhujiecha, Jiegu Lian, Jiegu Hernandia, etc. It is a commonly used traditional Chinese medicine recorded in Chinese Pharmacopoeia. Sarcandra glabra contains various flavonoids, terpenes, coumarins, organic acids, volatile oils, polysaccharides and other chemical components.

[0007] Currently, there is no report on the treatment of psoriasis and the reduction of the symptoms of psoriasis recurrence by Sarcandra glabra polysaccharide. SUMMARY

[0008] To solve the above problems, the application provides the use of Sarcandra glabra polysaccharide in the preparation of a medicine for preventing and / or treating psoriasis, especially reducing the symptoms of psoriasis recurrence.

[0009] The object of the application is achieved by the following technical scheme:

[0010] The use of Sarcandra glabra polysaccharide in the preparation of a medicine for preventing and / or treating psoriasis and delaying the recurrence of psoriasis.

[0011] The polysaccharide described above can reduce inflammation caused by psoriasis, decrease the expression of memory T cells in the skin lesions of psoriasis relapse model mice, and alleviate the symptoms of psoriasis in relapse model mice.

[0012] The extraction and separation method of the polysaccharide of *Hypericum sarmentosum* can follow the existing technology and is not particularly limited; it generally includes steps such as defatting, extraction, alcohol precipitation, washing, and dialysis.

[0013] Specifically, the extraction and separation method of the polysaccharide from *Hypericum sarmentosum* includes the following steps:

[0014] After drying and pulverizing *Hedyotis diffusa*, it is defatted 3-5 times with 3-10 times its volume of 60-100% ethanol. The residue is then extracted with water at 60-110℃ for 0.5-8 hours. The extraction is repeated several times. The extracts are combined, filtered, concentrated, and precipitated with 70% or higher ethanol solution. The precipitate is allowed to stand at 4-30℃ for 6-96 hours. The precipitate is collected by filtration, washed with organic solvent, reconstituted with water, and then dialyzed to obtain a solution containing *Hedyotis diffusa* polysaccharide. *Hedyotis diffusa* polysaccharide is then freeze-dried.

[0015] The organic solvent is preferably anhydrous ethanol, acetone, or diethyl ether.

[0016] A polysaccharide containing mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose;

[0017] Preferably, the molar mass ratio of mannose, rhamnose, galacturonic acid, glucose, galactose and arabinose is 5.3:5.8:3.6:58.1:19.7:1.3.

[0018] The molecular weight of the polysaccharide mentioned above is mainly distributed in the range of 6kDa-700kDa;

[0019] The polysaccharide of *Hypericum sarmentosum*, as determined by the sulfuric acid-phenol method, with glucose as a reference standard, has a sugar content of 60-96%.

[0020] A polysaccharide preparation containing *Hylocereus unicinctus* polysaccharide and a pharmaceutically acceptable carrier;

[0021] The pharmaceutically acceptable carrier includes one or more of the following: surfactants, excipients, stabilizers, suspending agents, isotonic agents, preservatives, fillers, disintegrants, antioxidants, emulsifiers, coating agents, binders, lubricants, or flavoring agents.

[0022] The dosage form of the preparation is powder, tablet, granule, capsule, oral liquid, or injection; the oral liquid is solution, syrup, emulsion, or suspension.

[0023] The lungixingfeng polysaccharide and / or lungixingfeng polysaccharide preparation can be applied to the preparation of a medicine for preventing and / or treating psoriasis and delaying recurrence of psoriasis.

[0024] The present application has the following advantages and effects relative to the prior art:

[0025] The present application provides application of lungixingfeng polysaccharide in preparation of a medicine for preventing and / or treating psoriasis and delaying recurrence of psoriasis. The present application finds that lungixingfeng polysaccharide can significantly improve the skin damage degree of a mouse of a psoriasis recurrence model induced by imiquimod (IMQ) at intervals, reduce the erythema, thickening and scales of the skin of the mouse of the psoriasis recurrence model induced by IMQ at intervals, and reduce the expression of memory T cells at a skin lesion of the mouse of the psoriasis recurrence model. Therefore, lungixingfeng polysaccharide has a great application prospect in preparation of a medicine for preventing and / or treating psoriasis, and can especially reduce recurrence symptoms of psoriasis. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 is the treatment effect of lungixingfeng polysaccharide on psoriasis (n=6); wherein, MTX: positive control group; SGP-L: lungixingfeng polysaccharide low-dose group; SGP-H: lungixingfeng polysaccharide high-dose group.

[0027] Figure 2 is the effect of lungixingfeng polysaccharide on a skin lesion of a mouse of a psoriasis recurrence model.

[0028] Figure 3 is a comparison of PASI scores of skin of a mouse of a psoriasis recurrence model by lungixingfeng polysaccharide; wherein, compared with the Control group, ## P<0.01; compared with the 2IMQ group, ** P<0.01, * P<0.05; compared with the MTX-2IMQ group, Δ P<0.05.

[0029] Figure 4 is a comparison of skin pathology of a mouse of a psoriasis recurrence model by lungixingfeng polysaccharide.

[0030] Figure 5 is the effect of lungixingfeng polysaccharide on the expression proportion of memory T cells of skin of a mouse of a psoriasis recurrence model.

[0031] Figure 6 is a molecular weight distribution diagram of lungixingfeng polysaccharide.

[0032] Figure 7 is an HPLC spectrum diagram of a monosaccharide mixed standard (upper) and a monosaccharide composition of lungixingfeng polysaccharide (lower); wherein, 1: Man, 2: Rha, 3: GluA, 4: GalA, 5: Glu, 6: Gal, 7: Ara.

[0033] Figure 8 is the infrared spectrum of SGP. DETAILED DESCRIPTION

[0034] The application will be further described in conjunction with the examples and drawings, but the embodiments of the application are not limited thereto.

[0035] Example 1

[0036] The extraction of SGP includes the following steps:

[0037] Defatting: 1 kg of SGP decoction pieces (batch number: 20210124, Kangmei Pharmaceutical Co., Ltd.) was weighed, ground into powder, and passed through a 10-mesh sieve. The powder was added to a 10 L round-bottom flask, soaked in 4 times 75% ethanol for 1 h, and then boiled and defatted for 1.5 h using an electronic temperature control heating jacket. The process was repeated twice. After the extraction was completed, the filtrate was collected by filtering through 4 layers of gauze, and the filter residue was dried and sealed for storage. The defatted SGP filter residue was weighed, and the defatting rate was 14.47%.

[0038] Extraction: 250 g of defatted SGP powder was weighed and placed in a 10 L round-bottom flask. Water was added at a material-to-liquid ratio of 1:20, and then boiled and extracted for 1 h using an electronic temperature control heating jacket. After extraction, the filtrate was collected by filtering through 4 layers of gauze, and the filter residue was collected. The extraction process was repeated 3 times, and the filtrate was combined and concentrated under reduced pressure. The concentrated solution was diluted to 250 mL, and then ethanol was added to a final concentration of 80% for alcohol precipitation. The solution was allowed to stand at room temperature for 24 h. The precipitated part was the polysaccharide component, which was brownish in color, and the yield was 8.68%.

[0039] Impurity removal: The precipitate (containing a small amount of ethanol) was centrifuged (5000 g, 10 min) in a centrifuge, and the residual ethanol in the upper layer was discarded. The precipitate was then washed with anhydrous ethanol, acetone, and diethyl ether, and the polysaccharide was collected after repeated washing. The dissolved SGP was transferred to a plastic bowl, sealed, and stored in a -80°C freezer. The SGP was freeze-dried, weighed, and sealed for storage.

[0040] Decolorization: An appropriate amount of SGP was dissolved in ultrapure water to prepare a 10 mg / mL polysaccharide solution. A macroporous resin D101 was added at a volume of 10% of the polysaccharide solution, and the mixture was shaken at 180 rpm in a 50°C constant-temperature shaking table for 45 min. The decolorized polysaccharide solution was freeze-dried to obtain SGP. The SGP was yellow-brown in color, the decolorization rate was 52.30%, and the polysaccharide retention rate after decolorization was 80.20%.

[0041] Example 2

[0042] Therapeutic effect of SGP on imiquimod-induced psoriasis mice

[0043] Animal modeling and drug administration:

[0044] 6-week-old male BALB / c mice were purchased from Guangdong Weitong Lihua Experimental Animal Technology Co., Ltd. (Experimental Animal Production License No. SCXK (Guangdong) 2022-0063). The experimental animals were bred in the barrier environment animal room of Guangdong Provincial Hospital of Chinese Medicine, with room temperature of 20-25°C, relative humidity of 40%-70%, and 12h / 12h light-dark alternation. This experiment was approved by the Ethics Committee of Guangdong Provincial Hospital of Chinese Medicine (NO. 2021090). Thirty BALB / c mice were randomly divided into blank control group, model control group, positive control group (MTX, 1 mg / kg), low-dose SGP group (SGP-L, 100 mg / kg), and high-dose SGP group (SGP-H, 400 mg / kg) according to body weight, with 6 mice in each group. The back skin of the mice was selected as the experimental observation area, and the long hair on the back was shaved with a razor before the experiment to expose a 2 cm x 3 cm hairless area. Except for the blank control group, 62.5 mg of imiquimod (IMQ) cream was applied to the exposed area of the back of the mice in the other groups, and continuous administration was performed for 7 days. Daily photographs were taken, and the psoriasis area and severity index (PASI) was scored. Erythema, scales, and thickness were each scored from 0 to 4 (0, none; 1, mild; 2, moderate; 3, severe; 4, extremely severe), and the total score was obtained by summing the scores of the three factors.

[0045] Experimental results: SGP can alleviate the symptoms of psoriasis in mice induced by imiquimod

[0046] The skin of the mice in the blank control group was smooth and delicate. The mice in the model control group showed significant scaling, erythema, and thickening at the skin lesions, indicating successful modeling. Compared with the model control group, the low- and high-dose SGP groups significantly improved the redness, scaling, and thickening of the mouse skin. This suggests that SGP has the potential to treat psoriasis. Figure 1 .

[0047] See Table 1. Compared with the blank control group, the PASI score of the model control group was significantly increased. Compared with the model control group, the PASI scores of the low-dose SGP group, the high-dose SGP group, and the positive control group were reduced by 56.36%, 60.00%, and 32.73%, respectively (P < 0.05, 0.01). Among them, the PASI score of the low-dose SGP group decreased the most, but there was no significant difference between the low- and high-dose groups. Compared with the positive control group, the PASI score of the high-dose SGP group was significantly reduced (P < 0.05). The above results indicate that SGP has a good therapeutic effect on psoriasis.

[0048] Table 1 Comparison of the effect of SGP on the PASI score of psoriasis mice n = 6

[0049]

[0050] Note: compared with the blank control group, ## P < 0.01; compared with the model control group, ** P < 0.01, * P < 0.05; compared with MTX, Δ P < 0.05.

[0051] Example 3

[0052] Effect of SGP on the recurrence of psoriasis induced by interval induction of imiquimod

[0053] Animal modeling and drug administration:

[0054] 6-week-old male BALB / c mice were purchased from Guangdong Weitong Lihua Experimental Animal Technology Co., Ltd. (Experimental Animal Production License No. SCXK (Guangdong) 2022-0063). The experimental animals were bred in the barrier environment animal room of Guangdong Provincial Hospital of Traditional Chinese Medicine, with room temperature of 20-25°C, relative humidity of 40%-70%, and 12h / 12h light / dark alternation. This experiment was approved by the Ethics Committee of Guangdong Provincial Hospital of Traditional Chinese Medicine (NO. 2021090). Forty BALB / c mice were randomly divided into normal control group (Control), model group (2IMQ), positive control group (MTX-2IMQ), SGP low-dose group (SGP-L-2IMQ, 100 mg / kg), and SGP high-dose group (SGP-H-2IMQ, 400 mg / kg), a total of 5 groups, 8 mice in each group. The initial modeling and drug administration methods of the experimental animals were the same as those in Example 11. After 14 days, except for the Control group, the mice in the other groups were smeared with a low-dose (20.8 mg) of imiquimod cream on the back for 7 consecutive days to induce the recurrence of psoriasis. Daily photography was performed, and the psoriasis area and severity index (PASI) was scored. Erythema, scales, and infiltration thickness were each scored from 0 to 4 (0, none; 1, mild; 2, moderate; 3, severe; 4, extremely severe), and the total score was obtained by adding the scores of the three indicators. HE staining was used to evaluate the pathological changes, and flow cytometry was used to investigate the expression ratio of CD8+CD103+ in mouse T lymphocytes in the mouse skin.

[0055] Experimental results:

[0056] Effect of SGP on psoriasis lesions in mice

[0057] Compared with the control group, the 2IMQ psoriasis relapse model mice showed obvious erythema, scaling, and thickening at the skin lesions, indicating successful modeling. Compared with the 2IMQ group, MTX-2IMQ, SGP-L-2IMQ, and SGP-H-2IMQ improved erythema, scaling, and thickening to varying degrees, as detailed in [link to relevant documentation]. Figure 2 .

[0058] Compared with the control group, the PASI score of the skin lesions in the relapse model group (2IMQ) mice was significantly increased. Figure 3 Compared with the 2IMQ group, the PASI scores of the skin lesions in mice in the MTX-2IMQ group, SGP-L-2IMQ group, and SGP-H-2IMQ group decreased by 30.00%, 49.00%, and 58.75%, respectively (P<0.05 or 0.01); compared with the MTX-2IMQ group, the PASI scores of the skin lesions in mice in the SGP-H-2IMQ group were significantly reduced (P<0.05). These results indicate that *Echinops latifolius* polysaccharide can alleviate the skin lesion symptoms in mice with psoriasis relapse and has good therapeutic potential.

[0059] Pathological comparison Figure 4 Compared with the Control group, the 2IMQ group showed increased epidermal thickness, elongated epidermal ridges, and the presence of parakeratotic cells in the stratum corneum, with significant thickening of the stratum spinosum, indicating successful model establishment. Compared with the 2IMQ group, the *Hypericum sibiricum* polysaccharide group significantly reduced skin thickness and elongated epidermal ridges. These results indicate that *Hypericum sibiricum* polysaccharide improves the pathological changes in skin lesions in mice with a psoriasis relapse model.

[0060] Effect of *Hypericum perforatum* polysaccharide on the proportion of CD8+CD103+TRM expression in skin memory T cells of psoriasis relapse mice:

[0061] Flow cytometry results are shown in Figure 5 See Table 2. Compared with the Control group, the proportion of CD8+CD103+ T cells in the skin lesions of the 2IMQ group was significantly increased, with an increase of 87.71% (P < 0.01). Compared with the 2IMQ group, the proportion of CD8+CD103+ T cells in the skin lesions of the SGP-H-2IMQ group and the SGP-L-2IMQ group decreased by 42.92% and 62.98%, respectively (P < 0.05, 0.01), while the proportion of CD8+CD103+ T cells in the skin lesions of the MTX-2IMQ group decreased by 16.06%, with no statistically significant differences. These results indicate that the mechanism by which *Rhizoma Scutellariae* polysaccharide delays psoriasis recurrence involves reducing the proportion of CD8+CD103+ T cells in the skin, while the positive control drug methotrexate had no significant effect on the expression ratio of CD8+CD103+ T cells.

[0062] Table 2. Effects of *Hypericum perforatum* polysaccharide on the expression ratio of skin memory T cells in mice with recurrent psoriasis.

[0063]

[0064] Note: compared with the Control group, ## P<0.01, compared with the 2IMQ group, * P<0.05, ** P<0.01

[0065] Conclusion: Argyreia Nuxvomica polysaccharide can significantly improve the symptoms of psoriasis-like skin lesions induced by imiquimod (IMQ), and can significantly reduce the degree of skin damage in the relapse model of psoriasis in mice induced by IMQ interval, and reduce the expression of memory T cells in the skin lesions of the relapse model of psoriasis in mice, therefore, Argyreia Nuxvomica polysaccharide has great application prospect in the preparation of drugs for preventing and / or treating psoriasis, especially in reducing the recurrence of psoriasis.

[0066] Example 4

[0067] Description of the physicochemical properties of Argyreia Nuxvomica polysaccharide

[0068] Sugar content of Argyreia Nuxvomica polysaccharide

[0069] The Argyreia Nuxvomica polysaccharide obtained in Example 1 was weighed and dissolved in ultrapure water to prepare a polysaccharide solution of 1 mg / ml. 200 μL of the polysaccharide solution was taken in a glass test tube, 100 μL of 5% phenol solution was added and mixed thoroughly, and then 500 μL of concentrated sulfuric acid was quickly added to cause vigorous reaction. After the system cooled at room temperature, the absorbance was measured at 490 nm on a microplate reader. The sugar content standard was prepared according to its monosaccharide composition, and was gradient diluted to concentrations of 1000, 500, 250, 125, 62.5, 31.25, 15.625, and 0 μg / mL, respectively. The standard curve was plotted with sugar content as the abscissa and absorbance as the ordinate. The calculated sugar content of the polysaccharide was 76.24%.

[0070] Determination of uronic acid content of Argyreia Nuxvomica polysaccharide

[0071] The uronic acid content of the polysaccharide was quantitatively analyzed by the m-hydroxydiphenyl method: D-galacturonic acid standard was accurately weighed, and distilled water was used to prepare a 1.0 mg / mL stock solution. The standard solution was gradient diluted to prepare a series of standard solutions of 3.9-250 μg / mL. 100 μL of the standard solution or sample solution (polysaccharide solution diluted to 100 μg / mL) was taken, 600 μL of sodium tetraborate-sulfuric acid mixed solution was added under ice bath condition, and then vortexed and mixed thoroughly. After heating in a boiling water bath for 12 min, the ice bath was cooled, 10 μL of m-hydroxydiphenyl methanol solution (1.5 mg / mL) was added, and the color was developed for 10 min in the dark.

[0072] Determination of protein content of Argyreia Nuxvomica polysaccharide

[0073] The content of protein in each sample was determined by Bradford method. Bradford reagent was prepared: Coomassie brilliant blue G-250 100 mg, dissolved in 95% ethanol 50 mL, add 85% phosphoric acid 100 mL, distilled water to 200 mL, filter and store at 4°C, which is the stock solution. Before use, dilute the stock solution 10 times with distilled water to get the reagent application solution. Accurately weigh 50 mg of bovine serum albumin, dissolve in 0.015 mol / L PBS 50 mL (concentration is 1000 mg / L) to make protein standard solution, dilute to 7 concentrations, the concentrations are 1000, 500, 250, 125, 62.5, 31.25, 15.63, 7.81 μg / mL, draw the protein content standard curve. Weigh an appropriate amount of SGP, dilute the polysaccharide to 500 μg / mL with ultrapure water, and calculate the protein content by absorbance. Protein content (%) = c x D / m x 100%, where c is the measured concentration of uronic acid (mg / mL), D is the dilution factor, and m is the mass of the sample being tested (mg). The protein content obtained by this method is 7.78%.

[0074] Molecular weight of SGP

[0075] Accurately weigh the dextran standard of different molecular weight (1 kDa-3690 kDa). Prepare a standard solution with a concentration of 1.0 mg / mL by dissolving in distilled water, filter through a 0.22 μm water filter membrane, and analyze using an Agilent 1260 high performance liquid chromatography system equipped with a differential refractive index detector. The chromatographic conditions are as follows: column type TSK G-5000PWXL, mobile phase ultrapure water, flow rate 0.4 mL / min, column temperature 40°C. Detector temperature 40°C, injection volume 10 μL. Establish a standard curve of the linear relationship between the log of the weight average molecular weight (log Mw) and the retention time by HPGPC system. The sample of SGP to be tested is pretreated in the same way and detected under the same chromatographic conditions. Evaluate the sample homogeneity by chromatographic peak shape, record the characteristic retention time, and calculate the molecular weight distribution parameters of the polysaccharide using the standard curve equation. As shown in the figure, the SGP has a wide distribution characteristic Figure 1 Figure 6 The molecular weight distribution range of SGP is 6-700 kDa, mainly divided into two parts, peak I with a peak molecular weight of 534 kDa, accounting for 33.24%, and peak II with a peak molecular weight of 73.2 kDa, accounting for 42.03%.

[0076] Monosaccharide composition of SGP:

[0077] ​The complete acid hydrolysis-PMP pre-column derivatization method was used for monosaccharide composition analysis: 8.0 mg of polysaccharide sample was precisely weighed, and 2 ml of 2M TFA solution was added for hydrolysis, and methanol was used for washing and evaporation to remove trace amounts of trifluoroacetic acid, and finally 800 μL of ultrapure water was used for reconstitution; 10.00 mg of each of seven monosaccharide standard samples, namely arabinose (Ara), glucose (Glc), rhamnose (Rha), mannose (Man), galactose (Gal), galacturonic acid (GalA), and glucuronic acid (GluA), was precisely weighed and mixed, and distilled water was used to prepare a 2.0 mg / mL stock solution, which was gradient diluted to 0-500 μg / mL mixed standard working solution; 500 μL of sample solution or standard solution was accurately transferred, and equal volumes of 0.3M NaOH and 0.6M PMP methanol solution were added, and derivatization was carried out at 70°C for 100 min. After the reaction was terminated, it was neutralized with 0.3M HCl, extracted with chloroform for 3 times (10 000×g centrifugation for 10 min), and the water phase was collected and filtered through a 0.22 μm filter for testing. An Agilent 1260HPLC system was used, a Diamonsil C18 chromatographic column (250mm×4.6mm, 5μm) was used, the mobile phase was 50mM phosphate buffer (pH6.83)-acetonitrile (83:17), the column temperature was 37°C, the detection wavelength was 254nm, and the injection volume was 10μL. The retention time was used for qualitative analysis, and the peak area was used for quantitative calculation of the molar ratio of each monosaccharide.

[0078] The determination results are shown in Figure 7 . The polysaccharide of Argyi was mainly composed of six monosaccharides, namely mannose, rhamnose, galacturonic acid, glucose, galactose and arabinose. The peak areas of the above monosaccharides were calculated by substituting the standard curve, and the molar percentages of them are shown in Table 3.

[0079] Table 3 Monosaccharide composition of Argyi polysaccharide

[0080]

[0081]

[0082] Example 5

[0083] A small amount of polysaccharide sample obtained in Example 1 was uniformly spotted on the surface of zinc selenide crystal, and after tabletting by Fourier transform infrared spectrometer (FT-IR), the infrared absorption spectrum was collected in the range of 500-4000 cm -1 .

[0084] The infrared spectrum can quickly analyze the sugar ring configuration, glycosidic bond type and substituent characteristics of polysaccharide through the vibration information of characteristic functional groups. The infrared spectrum of Argyi polysaccharide Figure 8 ) is 3326 cm -1A broad and strong absorption peak can be observed at 2925.10 cm⁻¹, which is a characteristic absorption peak of polysaccharides, reflecting OH stretching vibration and indicating the presence of intermolecular hydrogen bonds and hydroxyl groups; -1 The CH stretching vibration peak at 1417.52 cm⁻¹, and the peak at 1417.52 cm⁻¹. -1 The absorption peak at the CH angular vibration reflects the methylene structure of the sugar ring; 1748.01 cm⁻¹ -1 1606.82cm -1 (C=O) and 1417.00cm -1 (COH) indicates the possible presence of acidic groups or esterification modifications, i.e., uronic acid; 1200-1000 cm -1 The strong absorption band represents the COC glycosidic bond, binding at 600.05 cm⁻¹. -1 The presence of a strong absorption peak indicates that the sugar mainly exists in the form of furanose.

[0085] Example 6

[0086] Preparation of Polysaccharide Tablets for Rhizoma Sinapis Albae: Weigh 200g of Rhizoma Sinapis Albae polysaccharide raw material, 88g of lactose, and 9g of sodium hydroxymethyl starch, mix them, and pass them through a 120-mesh sieve three times to ensure even mixing. Add an appropriate amount of binder to make a soft mass, pass the soft mass through an 18-mesh sieve, dry the wet granules in an oven at 50℃ for 1 hour, granulate them through an 18-mesh sieve, add 3g of magnesium stearate and sodium hydroxymethyl starch (25% of the prescribed amount), mix well, and compress into tablets to obtain the final product.

[0087] Example 7

[0088] Preparation of Polysaccharide Capsules for Swelling Joints: Weigh 180g of Polysaccharide raw material, add 50g of starch and 50g of dextrin, mix well, use 90% ethanol as a wetting agent for wet granulation, vacuum dry at 50℃, granulate, pass through a 40-mesh sieve and a 200-mesh sieve to remove fine powder, and fill No. 3 capsules in an environment with humidity below 60% to obtain the product.

[0089] Example 8

[0090] Preparation of Polysaccharide Granules: 200g of β-cyclodextrin was dissolved in 200g of water and heated and stirred until dissolved; 40g of Polysaccharide Granules was dissolved in 50g of anhydrous ethanol and then slowly added dropwise to the β-cyclodextrin aqueous solution while stirring; after cooling, the granules were filtered to obtain crystals, washed, dried, pulverized, and passed through a 40-mesh sieve to obtain the final product.

[0091] Example 9

[0092] Preparation of microemulsion of polysaccharide of jingjie-feng: take 2.5g of Labrafil M1944Cs, 15g of polyoxyethylene hydrogenated castor oil EL35 and 5g of Transcutol HP, stir and mix, add 5g of polysaccharide of jingjie-feng, continue to stir, add 22.5g of distilled water, continue to stir at a constant speed to obtain a primary emulsion, and then ultrasonic treatment to form a light yellow transparent O / W type traditional Chinese medicine component combined microemulsion, and the preparation is completed.

[0093] Example 10

[0094] Preparation of emulsion of polysaccharide of jingjie-feng: according to the above prescription, take 10g of Labrafil M1944Cs, 12g of polyoxyethylene 40 hydrogenated castor oil (RH40), 4g of Transcutol HP, stir and mix, add 4g of polysaccharide of jingjie-feng, continue to stir, and then add 20g of distilled water after the drug is completely dissolved, continue to stir at a constant speed to obtain a primary emulsion, and then ultrasonic treatment to obtain an opaque emulsion, and the preparation is completed.

[0095] Example 11

[0096] Preparation of polysaccharide of jingjie-feng cream: take 6g of octadecanol, 4g of white vaseline, 3g of glyceryl monostearate as the oil phase, and 1.25g of Tween 80, 3.45g of glycerol, 0.05g of nipagin ethyl, and 29.75g of distilled water as the water phase, heat the oil phase and the water phase in a 80℃ water bath respectively, and then slowly pour the water phase into the oil phase while stirring, mix uniformly, and finally add 2.5g of polysaccharide of jingjie-feng raw material, continue to stir until cooling, and the preparation is completed.

[0097] Example 12

[0098] Preparation of polysaccharide of jingjie-feng gel: take 0.4g of carbomer 940, 10g of glycerol, 10g of anhydrous ethanol, and distilled water, stir and mix uniformly, and then place overnight, add 2.5g of polysaccharide of jingjie-feng, stir and mix uniformly, and then add 2g of azone, 1g of Tween-80, distilled water to 100g, adjust the pH to 5 with triethanolamine, add 0.1g of nipagin ethyl, stir and mix thoroughly, and the preparation is completed.

[0099] 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 should be equivalent replacement methods, and all are included in the protection scope of the present application.

Claims

1. Use of a polysaccharide of Sarcandra glabra in the preparation of a medicine for preventing and / or treating psoriasis and delaying recurrence of psoriasis.

2. Use according to claim 1, characterized in that: The extraction and separation method of the polysaccharide of Sarcandra glabra comprises the steps of degreasing, extraction, alcohol precipitation, washing, and dialysis.

3. Use according to claim 1, characterized in that: The extraction and separation method of the polysaccharide of Sarcandra glabra comprises the following steps: After the Sarcandra glabra is dried and crushed, 3-10 times the volume of 60-100% ethanol is added for degreasing 3-5 times, the residue is added with water, and extracted at 60-110°C for 0.5-8h, and the extraction is repeated for several times, the combined extract is filtered, concentrated, and added with 70% or more ethanol solution for precipitation, and then placed at 4-30°C for 6-96h; the precipitate is collected by filtration, washed with an organic solvent, redissolved with water, and then dialyzed to obtain a solution containing the polysaccharide of Sarcandra glabra, which is freeze-dried to obtain the polysaccharide of Sarcandra glabra.

4. Use according to claim 3, characterized in that: The organic solvent is anhydrous ethanol, acetone, or diethyl ether.

5. A polysaccharide of Sasa bartramia, characterized by: The monosaccharides include mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose.

6. The Polysaccharide of Sarcandra glabra according to claim 5, characterized in that: The molar mass ratio of the mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose is 5.3:5.8:3.6:58.1:19.7:1.

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7. The Polysaccharide of Sarcandra glabra according to claim 5, characterized in that: The polysaccharide of Sarcandra glabra has a molecular weight of 6kDa-700kDa.

8. A preparation of polysaccharides of Sasa bartramia, characterized in that: The polysaccharide of Sarcandra glabra as claimed in any one of claims 5-7 and a pharmaceutically acceptable carrier.

9. The Argyi Monks' Clubmoss Pollen preparation of claim 8, characterized by: The pharmaceutically acceptable carrier comprises one or more of a surfactant, an excipient, a stabilizer, a suspending agent, an isotonic agent, a preservative, a filler, a disintegrating agent, an antioxidant, an emulsifying agent, a coating agent, a binder, a lubricant, or a flavoring agent.

10. Use of the polysaccharide of Sarcandra glabra as claimed in any one of claims 5-7 and / or the preparation of the polysaccharide of Sarcandra glabra as claimed in claim 8 or 9 in the preparation of a medicine for preventing and / or treating psoriasis and delaying recurrence of psoriasis.

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