Sulfated zizania aquatica polysaccharide as well as preparation method and application thereof in resisting non-small cell lung cancer

Abstract

The invention provides sulfated zizania aquatica polysaccharide. The sulfated zizania aquatica polysaccharide is 62.6 + / -2.66% in total sugar content, 5.53 + / -0.23% in uronic acid content, 15.1 + / -2.50% in sulfate group content and 1.52 in sulfate group substitution degree. The sulfated zizania aquatica polysaccharide is prepared from arabinose, galactose, glucose, galacturonic acid and glucuronic acid according to a molar ratio of 42.8: 22.2: 227: 22.2: 1.00: 4.03. The molecular weight of the sulfated zizania aquatica polysaccharide is 1.48 kDa to 24.1 kDa; the particle size of the sulfated zizania aquatica polysaccharide is 634 + / -17.6 nm, and the potential is-27.9 + / -1.10 mV. The sulfated zizania aquatica polysaccharide provided by the invention has remarkable non-small cell lung cancer resisting activity and can be used for preparing functional products such as oral liquid, tablets and capsules.

Description

Technical Field

[0001] The invention belongs to the field of natural product preparation and application, and specifically relates to sulfated wild rice stem polysaccharide and a preparation method thereof and application thereof in preventing non-small cell lung cancer. Background Art

[0002] Zizania latifolia is a perennial shallow-water herb of the genus Zizania in the Poaceae family. The spindle-shaped fat part of its tender stems stimulated by Zizania black powder can be eaten as a vegetable. It has a long history of cultivation in the Yangtze River Basin of my country and is one of the characteristic aquatic vegetables of the "Eight Water Immortals" in the south of the Yangtze River. At the same time, Zizania has a long history of medicinal use. "Compendium of Materia Medica" records that it "removes heat, quenches thirst, removes yellow eyes, promotes urination and defecation, stops heat dysentery, and relieves alcohol poisoning." Zizania is known as the "ginseng in water" and is rich in polysaccharides, proteins, amino acids, vitamins, trace elements, polyphenols, flavonoids and other nutrients and bioactive substances. Among them, Zizania polysaccharides have pharmacological activities such as anti-oxidation, enhancing the body's immunity, and protecting the liver, and are the main functional ingredients of Zizania (Journal of Food Science, 2023, 88: 94-108).

[0003] According to the difference in histopathological types, lung cancer is divided into two categories: non-small cell lung cancer and small cell lung cancer. Among them, non-small cell lung cancer has the highest incidence, accounting for 85%. Although great progress has been made in the early diagnosis and treatment of non-small cell lung cancer in recent years, the prognosis of patients with non-small cell lung cancer is still poor, and the five-year survival rate is still less than 20% (Biomedicine & Pharmacotherapy, 2018, 108: 130-136). Therefore, the development of functional foods and new drugs related to the prevention and treatment of non-small cell lung cancer has become one of the most concerned issues in the field of biomedicine.

[0004] At present, the research on Zizania latifolia polysaccharides focuses on extraction, separation, enhancement of immunity, anti-oxidation, composite preservative film, etc. (CN115678113A; LWT-Food Science and Technology, 2023, 173: 114332; International Journal of Biological Macromolecules, 2017, 95: 809-817; International Journal of Biological Macromolecules, 2018, 107: 882-890; Journal of Food Science, 2023, 88: 94-108; Agricultural Products Processing, 2023, 577: 20-26; Food Industry Science and Technology, 2015, 17: 211-214; Grain and Oils, 2023, 36: 133-138), and there are no reports on sulfation modification and anti-non-small cell lung cancer applications. The invention provides a preparation method of sulfated wild rice stem polysaccharide, structural characteristics and use thereof in treating non-small cell lung cancer, and provides a lead compound for the development of novel anti-non-small cell lung cancer drugs. Summary of the invention

[0005] The purpose of the present invention is to provide a sulfated wild rice stem polysaccharide, which has excellent anti-non-small cell lung cancer activity and can significantly (P<0.01) inhibit the viability of A549 human non-small cell lung cancer cells. After 72 hours of action at a concentration of 0.5 mg / mL, the cell survival rate is 15.74±2.82%.

[0006] According to a first aspect of the present invention, the present invention provides a sulfated wild rice polysaccharide, wherein the sulfated wild rice polysaccharide is obtained by sulfating wild rice polysaccharide with aminosulfonic acid; the sulfate group substitution degree of the sulfated wild rice polysaccharide is 0.62-1.52.

[0007] According to a second aspect of the present invention, the present invention provides a method for preparing sulfated wild rice polysaccharide, comprising the following steps: placing wild rice polysaccharide and aminosulfonic acid in an organic solvent, heating and reacting for a certain period of time; cooling to room temperature, quenching the reaction with an alkaline solution, adjusting the pH of the reaction solution to neutral, precipitating with ethanol to obtain a precipitate, re-dissolving the precipitate with distilled water, dialyzing, and freeze-drying to obtain sulfated wild rice polysaccharide with different degrees of substitution;

[0008] The specific steps may be: weighing 100 mg of Zizania latifolia polysaccharide (ZLP) and placing it in a round-bottom flask, adding a certain amount of aminosulfonic acid according to different mass ratios, then adding an appropriate amount of solvent (N,N-dimethylformamide) and a catalytic amount of urea, turning on a magnetic stirrer to fully mix them, and reacting them at 80° C. for 3 hours; after cooling to room temperature, adding a 10 wt % NaOH aqueous solution to quench the reaction, and adjusting the pH value to neutral; adding 3 times the volume of anhydrous ethanol in the reaction solution and precipitating at 4° C. for 12 hours; re-dissolving the precipitate with distilled water, dialyzing with distilled water 3 times, and freeze-drying to obtain ZLP with different sulfate substitution degrees;

[0009] Preferably, the mass ratio of the wild rice polysaccharide to aminosulfonic acid is 1:1 to 1:4, more preferably 1:2;

[0010] Preferably, the sulfate substitution degree of the sulfated wild rice stem polysaccharide is 0.62 to 1.52;

[0011] Preferably, the mass ratio of the water bamboo polysaccharide to aminosulfonic acid is 1:2, and the sulfated water bamboo polysaccharide obtained has a sulfate group substitution degree of 1.52. At this time, the prepared sulfated water bamboo polysaccharide has a total sugar content of 62.6±2.66%, a uronic acid content of 5.53±0.23%, a sulfate group content of 15.1±2.50%, and a sulfate group substitution degree of 1.52; the obtained sulfated water bamboo polysaccharide contains the following molar ratios of monosaccharides, arabinose, galactose, glucose, galacturonic acid, glucuronic acid = 42.8:22.2:227:22.2:1.00:4.03; the molecular weight of the sulfated water bamboo polysaccharide is 1.48-24.1 kDa; the particle size of the sulfated water bamboo polysaccharide is 634±17.6 nm, and the potential is -27.9±1.10 mV;

[0012] Preferably, the process for preparing the sulfated wild rice stem polysaccharide is a purified wild rice stem polysaccharide, and comprises the following steps:

[0013] 1) Extraction of crude polysaccharides from Zizania latifolia

[0014] Peel, wash and slice the fresh wild rice stem, dry at 60℃ to constant weight, crush and pass through a 60-mesh sieve to make wild rice stem powder for later use; add petroleum ether to degrease in the dark for 12 hours, and filter under reduced pressure to obtain the wild rice stem defatted powder. Weigh 10g of wild rice stem defatted powder and soak it in water for 4 hours at a liquid-to-solid ratio of 50ml:1g, then transfer it to an ultrasonic extractor (XH-2008D, Beijing Xianghu Technology) and extract it at 300W and 60℃ for 30 minutes. After cooling and filtering, the filtrate is concentrated to 1 / 5 of the original volume under reduced pressure; add 3 times the volume of anhydrous ethanol and precipitate at 4℃ for 12 hours. The precipitate is freeze-dried to obtain wild rice stem crude polysaccharide, and the extraction rate is 16.9±2.10%. Crude polysaccharide extraction rate (%) = m(g) / M(g)×100, where m is the mass of wild rice stem crude polysaccharide and M is the mass of wild rice stem defatted powder;

[0015] 2) Purification of crude polysaccharide from wild rice stem

[0016] The protein was removed by Sevag method three times (Sevag reagent was composed of n-butanol and chloroform in a volume ratio of 4:1); after removing the residual Sevag reagent, the solution was dialyzed three times (the molecular weight cutoff of the dialysis bag was 7000Da, the distilled water was dialyzed for 72 hours, and the water was changed every 12 hours); the dialysate was freeze-dried to obtain the deproteinized polysaccharide.

[0017] Weigh 100 mg of deproteinized polysaccharide and dissolve it in 20 mL of deionized water. Load it onto a diethylaminoethyl (DEAE) cellulose-52 chromatographic column (2.6 cm × 60 cm). Use 0.5 mol / L NaCl aqueous solution as the eluent, with a flow rate of 1 mL / min and 5 mL per tube. Use the phenol-sulfuric acid method to detect the eluent. After the polysaccharide elution is completed, combine the eluents and perform concentration and dialysis. Then freeze-dry to obtain the purified Zizania latifolia polysaccharides, named ZLP (Zizania latifolia polysaccharides). After deproteinization, dialysis, and DEAE column chromatography purification, the polysaccharide content of ZLP is 82.3±2.37%.

[0018] According to another aspect of the present invention, the present invention provides the use of the sulfated wild rice stem polysaccharide in preparing a functional product against non-small cell lung cancer; it can be used to prepare oral liquids, tablets, capsules and other preparations.

[0019] Preferably, the oral liquid contains sulfated wild rice stem polysaccharide, xylitol, sodium benzoate and water.

[0020] Preferably, the tablet contains sulfated wild rice stem polysaccharide, dextrin, starch, tartaric acid and magnesium stearate.

[0021] Preferably, the capsule contains sulfated wild rice stem polysaccharide and starch.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] ① The anti-non-small cell lung cancer activity of Zizania latifolia polysaccharide was significantly enhanced by sulfation modification. The sulfate substitution degree of sulfated Zizania latifolia polysaccharide was 0.62-1.52, especially when the sulfate substitution degree was 1.52, the anti-non-small cell lung cancer activity was the highest.

[0024] ② The sulfated wild rice polysaccharide provided by the present invention can significantly (P<0.01) inhibit the viability of A549 human non-small cell lung cancer cells after acting at a concentration of 0.5 mg / mL for 72 hours, with a cell survival rate of 15.74±2.82%, and the inhibitory effect is 3.3 times that of the unmodified wild rice polysaccharide. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Effects of 0.5 mg / mL ZLP and its sulfate on A549 cell viability at 48 h;

[0026] Figure 2 . Effects of 0.5 mg / mL ZLP and its sulfate on A549 cell viability at 72 h;

[0027] Figure 3 .Chromatogram of monosaccharide composition of SZLP-2 (peaks in the standard: 1-fucose; 2-rhamnose; 3-arabinose; 4-galactose; 5-glucose; 6-xylose; 7-mannose; 8-fructose; 9-galacturonic acid; 10-glucuronic acid);

[0028] Figure 4 .Molecular weight distribution diagram of SZLP-2;

[0029] Figure 5 .Infrared spectrum of SZLP-2;

[0030] Figure 6 .Scanning electron micrograph of SZLP-2. DETAILED DESCRIPTION

[0031] Zizania latifolia was collected from the Zizania latifolia planting base in Chefang Town, Wuzhong District, Suzhou City, Jiangsu Province; monosaccharide standards, T series glucan standards, etc. were purchased from Sinopharm Chemical Reagent Co., Ltd.; A549 human non-small cell lung cancer cell line ( CCL-185 TM ) was purchased from Nanjing Kebai Biotechnology Co., Ltd.; the molecular weight was determined by a Waters high performance liquid chromatograph, model 1525; the monosaccharide composition was determined by a Dionex ion chromatograph, model ICS-5000; the infrared spectrum was determined by a PerkinElmer Fourier transform infrared microscope system, model Spotlight 200i; the scanning electron microscope (SEM) was determined by a HITACHI ultra-high resolution field emission scanning electron microscope, model Regulus8100.

[0032] Example 1 Extraction of crude wild rice polysaccharide

[0033] Peel, wash and slice fresh wild rice stems, dry at 60℃ to constant weight, crush and pass through a 60-mesh sieve to make wild rice stem powder for later use; add petroleum ether to defat for 12 hours in dark, and filter under reduced pressure to obtain wild rice stem defatted powder. Weigh 10g of wild rice stem defatted powder and soak it in water for 4 hours according to a liquid-to-solid ratio of 50ml:1g, then transfer it to an ultrasonic extractor (XH-2008D, Beijing Xianghu Technology) and extract it for 30 minutes at 300W and 60℃. After cooling and filtering, the filtrate is concentrated to 1 / 5 of the original volume under reduced pressure; add 3 times the volume of anhydrous ethanol after concentration and precipitate at 4℃ for 12 hours, and the precipitate is freeze-dried to obtain wild rice stem crude polysaccharide, with an extraction rate of 16.9±2.10%.

[0034] Crude polysaccharide extraction rate (%) = m (g) / M (g) × 100, m is the mass of crude water bamboo shoot polysaccharide, and M is the mass of defatted water bamboo shoot powder.

[0035] Example 2 Purification of crude wild rice polysaccharide

[0036] The protein was removed by Sevag method for 3 times (Sevag reagent was composed of n-butanol and chloroform in a volume ratio of 4:1); after removing the residual Sevag reagent, the solution was dialyzed for 3 times (dialysis bag molecular weight cutoff was 7000Da, distilled water was dialyzed for 72h, and water was changed every 12h); the dialysate was freeze-dried to obtain the deproteinized polysaccharide. 100mg of deproteinized polysaccharide was weighed and dissolved in 20mL of deionized water, and loaded onto a diethylaminoethyl (DEAE) cellulose-52 chromatographic column (2.6cm×60cm); 0.5mol / L NaCl aqueous solution was used as the eluent, the flow rate was 1mL / min, and each tube was 5mL; the eluent was detected by phenol-sulfuric acid method, and after the polysaccharide elution was completed, the eluent was combined and concentrated, dialyzed, and freeze-dried to obtain the purified Zizania latifolia polysaccharide, named ZLP (Zizania latifolia polysaccharides). The polysaccharide content of ZLP after deproteinization, dialysis and purification by DEAE column chromatography was 82.3±2.37%.

[0037] Example 3 Preparation of Sulfated Zizania latifolia Polysaccharide

[0038] 100 mg of the purified ZLP prepared in Example 2 was weighed and placed in a round-bottom flask, and 100 mg, 200 mg and 400 mg of aminosulfonic acid (SA) were added respectively, and then an appropriate amount of solvent (N,N-dimethylformamide) and a catalytic amount of urea were added, and a magnetic stirrer was turned on to mix them thoroughly, and the reaction was carried out at 80° C. for 3 h; after cooling to room temperature, a 10 wt % NaOH aqueous solution was added to quench the reaction, and the pH value was adjusted to neutral; 3 times the volume of the reaction solution was added with anhydrous ethanol and precipitated at 4° C. for 1 h. 2h; the precipitate was reconstituted with distilled water, dialyzed with distilled water for 3 times (the specific operation steps are: the precipitate was reconstituted with 10-20mL of distilled water, and placed in a dialysis bag with a molecular weight cutoff of 7000Da; dialyzed in 500mL of distilled water for 12h, and repeated 3 times), and freeze-dried to obtain sulfated ZLP, which were named SZLP-1 (ZLP:SA=1:1), SZLP-2 (ZLP:SA=1:2) and SZLP-3 (ZLP:SA=1:4) respectively.

[0039] Example 4 Composition Analysis of Sulfated Zizania Lagenaria Polysaccharide

[0040] The total sugar content was determined by the phenol-sulfuric acid method; the uronic acid content was determined by the m-hydroxybiphenyl method; the sulfate content was determined by the barium chloride gelatin turbidimetry; the sulfate substitution degree (DS) = 1.62×S / (32-1.02×S), S is the percentage of sulfate; the potential and particle size were determined using Malvern's nanoparticle size and zeta potential distribution instrument, the instrument model is Zetasizer Nano ZS.

[0041] As shown in Table 1, as the amount of aminosulfonic acid (SA) increases, the total sugar content of sulfated ZLP decreases; the uronic acid content increases; the sulfate content and degree of substitution increase first and then decrease; the potential value decreases first and then increases; and the particle size decreases. In summary, when ZLP:SA=1:2, the sulfate content and degree of substitution are the largest, and the potential value is the lowest.

[0042] Table 1. Composition analysis results of ZLP and its sulfates

[0043]

[0044] Different superscript letters represent statistically significant differences, P < 0.05 or P < 0.01.

[0045] Example 5 Determination of Anti-non-small Cell Lung Cancer Activity of Sulfated ZLP

[0046] The survival rate of A549 cells was determined by MTT assay. Figure 1As shown in the results, after 48 hours of action at a concentration of 0.5 mg / mL (water system), SZLP-2 could significantly inhibit the growth of A549 cells compared with ZLP (P<0.05), and the survival rate of A549 cells was 60.16±6.45%. This indicates that sulfation modification can enhance the anti-non-small cell lung cancer activity of ZLP.

[0047] like Figure 2 As shown in the results, after 72 hours of action at a concentration of 0.5 mg / mL, all sulfates significantly inhibited the growth of A549 cells compared with ZLP (P<0.01). Among them, SZLP-2 had the highest inhibitory activity, with an A549 cell survival rate of 15.74±2.82%, which was significantly lower than that of SZLP-1 (P<0.01) and SZLP-3 (P<0.05). This further indicates that sulfation modification can significantly enhance the anti-NSCLC activity of ZLP, and is positively correlated with the sulfate substitution degree. SZLP-2 has the highest sulfate substitution degree (1.52) and the highest anti-NSCLC activity.

[0048] Example 6 Further Characterization of SZLP-2

[0049] Considering that SZLP-2 has high anticoagulant activity, its structure was further characterized.

[0050] Monosaccharide composition analysis: Weigh 5 mg of SZLP-2 into a 5 mL stoppered graduated test tube, add 1 mL of 2 mol / L trifluoroacetic acid, hydrolyze in an oven at 121°C for 2 h, distilled water to 50 mL, filter with a 0.45 μm microporous filter membrane for injection analysis, the injection volume is 20 μL. Instrument and chromatographic conditions: American Dionex ICS-5000 ion chromatograph, equipped with a pulsed amperometric detector, the chromatographic column is CarboPac PA20 (6.5 μm, 3 mm × 150 mm). Gradient elution is used, the mobile phase is: 98% water + 2% 250 mM NaOH (0-21 min); 93% water + 2% 250 mM NaOH + 5% 1 M NaAc (21.1-30 min); 20% water + 80% 250 mM NaOH (30.1-50 min). The flow rate is 0.5 mL / min.

[0051] Molecular weight analysis: SZLP-2 was prepared into a 2 mg / mL solution, and the sample was filtered with a 0.45 μm microporous filter membrane before injection. The injection volume was 20 μL. The analytical instrument was a Waters 1525 high performance liquid chromatograph, a 2414 differential refractive index detector and an Empower 3 workstation, and the chromatographic column was Ultrahydrogel TMLinear (2μm, 7.8mm×300mm). Using dextran of different molecular weights as standard sugars, a series of standard sugar solutions of different molecular weights were prepared, and a third-order calibration curve for polysaccharide molecular weight determination was established based on retention time and molecular weight values. The chromatographic conditions were as follows: mobile phase: 0.1M NaNO3; flow rate: 0.9mL / min; column temperature: 45℃.

[0052] Infrared spectrum analysis: 2 mg of dry SZLP-2 was mixed with 100 mg of dry potassium bromide, pressed into tablets, and then placed in an infrared spectrometer for measurement with a scanning range of 4000 cm -1 ~400cm -1 .

[0053] Scanning electron microscope analysis: Take SZLP-2, stick it on the stage of double-sided conductive adhesive with a cotton swab, and use an ear-cleaning bulb to blow away the polysaccharide powder that is not stuck. Place it in an ion sputtering instrument for foil spraying, and then put the treated samples into the scanning electron microscope in order, and observe their morphology after the instrument setting is stable.

[0054] like Figure 3 As shown, SZLP-2 was determined to be composed of arabinose, galactose, glucose, galacturonic acid, and glucuronic acid in a molar ratio of 42.8:22.2:227:22.2:1.00:4.03 by comparison with the standard.

[0055] like Figure 4 As shown, SZLP-2 is a heterogeneous polysaccharide, including two peaks (peak I and peak II) with a molecular weight ranging from 1.48 to 24.1 kDa.

[0056] like Figure 5 As shown in the infrared spectrum of SZLP-2, it can be observed that 3449 cm -1 There is a strong absorption peak near 2981cm, which is caused by the stretching vibration of -OH; -1 It is the asymmetric stretching vibration of methyl CH; 2909cm -1 It is the asymmetric stretching vibration of methylene CH; 1639cm -1 is the stretching vibration of carbonyl CO; 1398cm -1 CO stretching vibration; 1063cm -1 The absorption peak near 892cm is the skeleton vibration of COC glycosidic bond; -1 COS stretching vibration; 618cm -1 The absorption peak is the pyranose ring. Infrared spectrum analysis shows that SZLP-2 conforms to the structural characteristics of sulfur-containing polysaccharides.

[0057] like Figure 6As shown, at a magnification of ×10k, the surface of SZLP-2 is relatively rough and presents an irregular honeycomb shape.

[0058] Example 7 Preparation of SZLP-2 Oral Liquid

[0059] Take 1.0g SZLP-2 and add it into 100mL purified water, stir at room temperature until dissolved, add 0.2g xylitol and 0.1g sodium benzoate respectively, stir evenly, sterilize instantly in cans, bottle (10mL / bottle), and seal to obtain SZLP-2 oral solution.

[0060] Example 8 Preparation of SZLP-2 Tablets

[0061] Take 1.0g SZLP-2 and mix it evenly with 1.0g starch and 1.5g dextrin, add 0.1g tartaric acid and 2mL 50% ethanol, mix evenly and pass through an 18-mesh sieve; after drying at 60°C, pass through an 18-mesh sieve again, add 0.1g magnesium stearate, mix evenly, and press into tablets to obtain SZLP-2 tablets.

[0062] Example 9 Preparation of SZLP-2 Capsules

[0063] Take 1.0g SZLP-2, spray it with 95% ethanol in a ratio of 1:1, mix well, add 10% starch to make a soft material, granulate it through an 18-mesh sieve, dry it in an oven at 60℃, granulate it through an 18-mesh sieve again, and fill it into No. 3 empty capsules in an environment with a relative humidity of less than 65% to obtain SZLP-2 capsules.

[0064] 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 sulfated wild rice polysaccharide, wherein the sulfated wild rice polysaccharide is obtained by sulfating wild rice polysaccharide with aminosulfonic acid; the sulfate group substitution degree of the sulfated wild rice polysaccharide is 0.62 to 1.

52.

2. The sulfated wild rice stem polysaccharide according to claim 1, characterized in that: The sulfate substitution degree of the sulfated wild rice stem polysaccharide is 1.

52.

3. The sulfated wild rice polysaccharide according to claim 2, wherein the sulfated wild rice polysaccharide comprises monosaccharides in the following molar ratios: arabinose, galactose, glucose, galacturonic acid, glucuronic acid = 42.8:22.2:227:22.2:1.00:4.03; the total sugar content, the uronic acid content, and the sulfate content of the sulfated wild rice polysaccharide are 62.6±2.66%, 5.53±0.23%, and 15.1±2.50%; the molecular weight of the sulfated wild rice polysaccharide is 1.48-24.1 kDa; the particle size of the sulfated wild rice polysaccharide is 634±17.6 nm and the potential is -27.9±1.10 mV.

4. A method for preparing the sulfated wild rice polysaccharide according to claim 1, comprising the following steps: placing wild rice polysaccharide and aminosulfonic acid in an organic solvent and heating for a certain period of time; cooling to room temperature, quenching the reaction with an alkaline solution, adjusting the pH of the reaction solution to neutral, precipitating with ethanol to obtain a precipitate, re-dissolving the precipitate with distilled water, dialyzing, and freeze-drying to obtain sulfated wild rice polysaccharides with different degrees of substitution.

5. The method according to claim 4, characterized in that: The mass ratio of the wild rice stem polysaccharide to aminosulfonic acid is 1:1 to 1:

4.

6. The method according to claim 5, characterized in that: The mass ratio of the wild rice stem polysaccharide to aminosulfonic acid is 1:2, and the sulfate substitution degree of the obtained sulfated wild rice stem polysaccharide is 1.

52.

7. The method according to claim 6, characterized in that: In the process of preparing the sulfated wild rice stem polysaccharide, the wild rice stem polysaccharide is purified wild rice stem polysaccharide, which is prepared by the following steps: 1) Extraction of crude polysaccharides from Zizania latifolia The fresh wild rice stem is peeled, washed, sliced, dried to constant weight, crushed and sieved to make wild rice stem powder for later use; petroleum ether is added for defatting, and the defatted wild rice stem powder is obtained by vacuum filtration; the water is weighed and soaked, and then transferred to an ultrasonic extractor for extraction; the filtrate is filtered after cooling, and the filtrate is concentrated under reduced pressure; anhydrous ethanol is added for precipitation, and the precipitate is freeze-dried to obtain the crude wild rice stem polysaccharide; 2) Purification of crude polysaccharide from wild rice stem Deproteinization is performed by Sevag method, residual Sevag reagent is removed and dialyzed, and the dialyzate is freeze-dried to obtain deproteinized polysaccharide; the deproteinized polysaccharide is weighed and dissolved in deionized water, and loaded onto a diethylaminoethylcellulose-52 chromatographic column; an aqueous NaCl solution is used as an eluent for elution; the eluent is detected by phenol-sulfuric acid method, and after the polysaccharide elution is completed, the eluent is combined and concentrated, dialyzed, and freeze-dried to obtain purified wild rice stem polysaccharide; The obtained sulfated water bamboo polysaccharide contains monosaccharides in the following molar ratios: arabinose, galactose, glucose, galacturonic acid, glucuronic acid = 42.8:22.2:227:22.2:1.00:4.03; the total sugar content of the sulfated water bamboo polysaccharide is 62.6±2.66%, the uronic acid content is 5.53±0.23%, and the sulfate content is 15.1±2.50%; the molecular weight of the sulfated water bamboo polysaccharide is 1.48-24.1 kDa; the particle size of the sulfated water bamboo polysaccharide is 634±17.6 nm, and the potential is -27.9±1.10 mV.

8. Use of sulfated wild rice stem polysaccharide, wherein the sulfated wild rice stem polysaccharide is selected from claims 1 to 3 or prepared by any preparation method in claims 4 to 7, and is used for preparing an anti-non-small cell lung cancer drug.

9. The use according to claim 8, characterized in that The dosage form of the anti-non-small cell lung cancer drug is oral liquid, tablet or capsule.

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