An antitumor composition containing N-acetyl-D-glucosamine and a method for preparing the same

Abstract

The application discloses an anti-tumor N-acetyl-D-glucosamine-containing composition and a preparation method thereof in the field of anti-tumor preparations, and comprises the following components in parts by weight: functionalized N-acetyl-D-glucosamine 1-3 parts, and polypeptide modifier 3-5 parts. The application can play an anti-tumor role through the immune regulation function of N-acetyl-D-glucosamine, the sugar metabolism interference, the immune regulation and the apoptosis induction in three directions by the application of the polypeptide modifier and N-acetyl-D-glucosamine. N-acetyl-D-glucosamine (GlcNAc) is a kind of bioactive sugar molecule and can have the following effects in tumor microenvironment regulation, i.e. sugar metabolism interference: cancer cells usually rely on sugar metabolism (Warburg effect) for rapid growth, the functionalized N-acetyl-D-glucosamine can inhibit the proliferation of cancer cells by affecting the sugar metabolism of the cancer cells.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of antitumor preparations, and particularly relates to an antitumor composition containing N-acetyl-D-glucosamine and a preparation method thereof. BACKGROUND

[0002] N-acetyl-D-glucosamine (GlcNAc) is a derivative of glucose, widely exists in the cell wall components of human body, especially as a component of glycosaminoglycans and glycoproteins. It is found that N-acetyl-D-glucosamine and its derivatives not only play an important role in immune regulation, but also may have certain therapeutic effect against tumors. The inhibitory effect of N-acetyl-D-glucosamine on tumors is mainly achieved through the following mechanisms: immune regulation: N-acetyl-D-glucosamine can enhance the function of the immune system and improve the anti-tumor immune response. For example, it can enhance the phagocytic function of macrophages, promote the activity of natural killer cells (NK cells), and regulate the function of T cells, thereby enhancing the body's ability to recognize and eliminate tumor cells; inhibition of tumor cell proliferation: studies have shown that N-acetyl-D-glucosamine can inhibit the proliferation of tumor cells by regulating the expression of cell cycle proteins. Specifically, it may affect certain signaling pathways (such as PI3K / Akt, MAPK, etc.) to reduce the division and proliferation of tumor cells; induction of tumor cell apoptosis: N-acetyl-D-glucosamine can also inhibit tumor growth by inducing tumor cell apoptosis. Studies have shown that it can activate apoptosis-related signaling pathways and increase the incidence of tumor cell death; inhibition of tumor angiogenesis: the growth and metastasis of tumors often depend on the formation of blood vessels, and N-acetyl-D-glucosamine has been shown in some studies to inhibit the formation of new blood vessels in tumors, thereby inhibiting the spread of tumors. In anti-tumor therapy, common side effects mainly come from the use of chemotherapy, radiotherapy and other anti-tumor drugs. The following are some common side effects: gastrointestinal side effects: chemotherapy drugs often cause nausea, vomiting, loss of appetite and diarrhea, etc. N-acetyl-D-glucosamine may help alleviate these discomforts through its immune regulation effect, especially its protective effect on the gastrointestinal tract; immune system suppression: chemotherapy and some targeted therapy drugs can suppress the immune system, increasing the risk of infection. N-acetyl-D-glucosamine can help enhance the function of the immune system and promote the activity of immune cells, thereby helping to alleviate the side effects of immune suppression; bone marrow suppression: many anti-tumor drugs can cause bone marrow suppression, leading to anemia, leukopenia or thrombocytopenia. By using N-acetyl-D-glucosamine in combination, it may help alleviate these adverse reactions, as N-acetyl-D-glucosamine can promote cell repair and regeneration; through reasonable combination, N-acetyl-D-glucosamine is expected to alleviate these common side effects in anti-tumor therapy, enhance treatment effect, and improve the quality of life of patients. SUMMARY

[0003] In view of the above, in order to overcome the defects of the prior art, the present application provides an anti-tumor N-acetyl-D-glucosamine-containing composition and a preparation method thereof. Through polypeptide modification and the application of N-acetyl-D-glucosamine, the immune regulation function of N-acetyl-D-glucosamine (GlcNAc) as a bioactive sugar molecule can be utilized in tumor microenvironment regulation. The possible effects of N-acetyl-D-glucosamine in tumor microenvironment regulation include sugar metabolism interference: cancer cells usually rely on sugar metabolism (Warburg effect) for rapid growth, and functionalized N-acetyl-D-glucosamine can inhibit the proliferation of cancer cells by affecting their sugar metabolism. The anti-tumor effect can be synergistically exerted through sugar metabolism interference, immune regulation, and apoptosis induction.

[0004] In order to achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows: the present application provides an anti-tumor N-acetyl-D-glucosamine-containing composition, and the preparation raw materials of the composition include the following components by weight: functionalized N-acetyl-D-glucosamine 1-3 parts, polypeptide modifier 3-5 parts;

[0005] Preferably, the preparation method of the functionalized N-acetyl-D-glucosamine specifically includes the following steps:

[0006] S1, dissolve D-glucosamine in dichloromethane, place it in a 0℃ environment, add triethylamine, stir and mix for 10 min, then slowly add chloroformic acid-2,2,2-trichloroethyl ester / dichloromethane solution, keep the temperature at 0℃, react for 30-50 min, then increase the reaction temperature to room temperature, continue to stir for 2-3 h, adjust the reaction pH to neutral with saturated sodium bicarbonate solution, extract with dichloromethane, wash the organic phase with saturated sodium chloride aqueous solution, dry the organic phase with anhydrous sodium sulfate, concentrate and purify to obtain N-Troc glucosamine;

[0007] Preferably, in step S1, the weight concentration of D-glucosamine in dichloromethane is 40-60 mg / mL;

[0008] Preferably, in step S1, the mass-volume ratio between D-glucosamine and triethylamine is 0.7-0.9 g / mL;

[0009] Preferably, in step S1, the mass-volume ratio between D-glucosamine and chloroformic acid-2,2,2-trichloroethyl ester is 1-1.2 g / mL;

[0010] S2, the N-Troc-amino glucose prepared in step S1 is dissolved in dichloromethane, and hydrobromic acid-acetic acid solution is added dropwise under ice water bath condition, stirring is carried out at a speed of 120-180 rpm, after 2-4 h of reaction, deionized water is added to quench the reaction, after the reaction system is neutralized to neutral with saturated sodium bicarbonate aqueous solution, dichloromethane is added for extraction, the organic phase is collected, washed with saturated sodium chloride aqueous solution, and the organic phase is dried with anhydrous sodium sulfate, concentrated and purified to obtain bromo-substituted-N-Troc-amino glucose;

[0011] Preferably, in step S2, the mass concentration of the N-Troc-amino glucose in dichloromethane is 20-40 mg / mL;

[0012] Preferably, in step S2, the volume ratio between hydrobromic acid and acetic acid in the hydrobromic acid-acetic acid solution is 1:2-3;

[0013] Preferably, in step S2, the mass-volume ratio between the N-Troc-amino glucose and the hydrobromic acid-acetic acid solution is 0.05-0.15 g / mL;

[0014] S3, the bromo-substituted-N-Troc-amino glucose prepared in step S2 is dissolved in DMF, and sodium azide is added and mixed uniformly, stirring is carried out at a speed of 200-300 rpm at room temperature, after 30-60 min of reaction, water is added to quench the reaction, ethyl acetate is added, the organic phase is collected, washed with saturated sodium chloride solution, the organic phase is separated, dried with anhydrous sodium sulfate, filtered, the filtrate is collected, concentrated and purified to obtain azido alpha-D-glucose;

[0015] Preferably, in step S3, the mass concentration of the bromo-substituted-N-Troc-amino glucose in DMF is 10-30 mg / mL;

[0016] Preferably, in step S3, the mass ratio between the bromo-substituted-N-Troc-amino glucose and sodium iodide is 5:2-4;

[0017] S4, the azido alpha-D-glucose prepared in step S3 is dissolved in anhydrous methanol, glacial acetic acid is added, mixed uniformly, zinc powder is added, and placed at 50-70°C for 2-4 h, filtered, the filtrate is collected, transferred to an ice water bath, acetic anhydride is added, stirring is carried out at a speed of 150-180 rpm, after 2-3 h of reaction, saturated sodium bicarbonate aqueous solution is added to adjust the pH to neutral, ethyl acetate is added for extraction, washed with saturated sodium chloride solution, the organic phase is separated, dried with anhydrous sodium sulfate, filtered, the filtrate is collected, concentrated and purified to obtain functionalized N-acetyl-D-amino glucose;

[0018] Preferably, in step S4, the mass concentration of the azido-α-D-glucose in the anhydrous methanol is 0.05-0.1 g / mL;

[0019] Preferably, in step S4, the volume ratio between the glacial acetic acid and the anhydrous methanol is 0.75-1.2:1;

[0020] Preferably, in step S4, the added mass of the zinc is 10-30% of the mass of the azido-α-D-glucose;

[0021] Preferably, in step S4, the mass-volume ratio between the azido-α-D-glucose and the acetic anhydride is 10-50 mg / mL;

[0022] Preferably, the method for preparing the polypeptide modifier specifically comprises the following steps:

[0023] A1, after the Rink Amide Resin resin is placed in DCM for soaking and swelling, the solvent is removed, the Rink Amide Resin resin is transferred to a pyridine / DMF solution, and after stirring for 5-10 min, the pyridine / DMF solvent is removed, and then new pyridine / DMF solvent is added for continued stirring for 10-15 min, and then the Rink Amide Resin resin is washed with DMF to obtain a deprotection group Rink Amide Resin resin;

[0024] Preferably, in step A1, the volume fraction of pyridine in the pyridine / DMF solution is 15%-25% in DMF;

[0025] A2, Fmoc-Pbf-arginine is dissolved in DMF, HBTU and HOBt are added, and after thorough mixing, DIEA is added, and stirring is continued at a speed of 150-180 rpm for 3-5 min, then the deprotection group Rink Amide Resin resin prepared in step A1 is added, and stirring is carried out at a speed of 200-300 rpm at room temperature for 1-2 h, then filtration is performed, and the Rink Amide Resin resin is collected and repeatedly washed with DMF to obtain a Rink Amide Resin resin-oligomeric arginine;

[0026] Preferably, in step A2, the mass concentration of the Fmoc-Pbf-arginine in the DMF is 30-50 mg / mL;

[0027] Preferably, in step A2, the mass ratio between the Fmoc-Pbf-arginine, HBTU and HOBt is 1.5-2:1:0.2-0.6;

[0028] Preferably, in step A2, the mass-volume ratio between the Fmoc-Pbf-arginine and the DIEA is 2.0-2.5 g / mL;

[0029] A3, dissolve the p-carboxybenzenesulfonamide in DMF, add dichlorosulfoxide dropwise under an ice water bath, stir at room temperature at a speed of 200-300 rpm overnight, after the reaction is completed by TLC detection, remove the solvent to obtain the intermediate product A, dissolve the intermediate product A in DMF, add it to the reaction system in step A2, add TEA to keep the pH of the reaction system at 8-9, after reacting at 25-40°C for 4-8 h, add ethyl acetate for extraction, collect the organic phase, wash with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain the Rink Amide Resin resin-oligarginine-benzenesulfonamide;

[0030] Preferably, in step A3, the mass concentration of the p-carboxybenzenesulfonamide in DMF is 40-50 mg / mL;

[0031] Preferably, in step A3, the mass-volume ratio between the p-carboxybenzenesulfonamide and dichlorosulfoxide is 0.2-0.25 g / mL;

[0032] A4, place the Rink Amide Resin resin-oligarginine-benzenesulfonamide prepared in step A3 in a TFA cleavage solution, stir at a speed of 150-180 rpm for 2-3 h, filter, collect the filtrate, add pre-cooled anhydrous ether for precipitation, centrifuge, discard the supernatant, collect the precipitate, dry to obtain the oligarginine-intermediate product A;

[0033] Preferably, in step A4, the volume of the TFA cleavage solution is 100-150 mL;

[0034] A5, dissolve 4-pentynoic acid in dichloromethane, place it in an ice water bath, add dichlorosulfoxide, raise the temperature to room temperature, stir at a speed of 200-300 rpm for 12-16 h, add potassium carbonate, transfer to ice water, add the oligarginine-benzenesulfonamide prepared in step A4, stir at a speed of 200-300 rpm at room temperature for 8-12 h, add ethyl acetate for extraction, collect the organic phase, wash with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain the polypeptide modifier;

[0035] Preferably, in step A5, the mass concentration of the 4-pentynoic acid in dichloromethane is 40-50 mg / mL;

[0036] Preferably, in step A5, the mass-volume ratio between the 4-pentynoic acid and the thionyl chloride is 0.1-0.15 g / mL:

[0037] Preferably, in step A5, the mass ratio between the 4-pentynoic acid and the potassium carbonate is 1:2.5-4;

[0038] The present application also provides a preparation method of an anti-tumor N-acetyl-D-glucosamine-containing composition, specifically comprising the following steps:

[0039] ① Functionalized N-acetyl-D-glucosamine and polypeptide modifiers are dissolved in a tert-butyl alcohol / water solution according to weight parts, sodium ascorbate and copper sulfate pentahydrate are added, and after stirring at 80-90℃ for 2-3h, ethyl acetate is added for extraction, the organic phase is collected, dried with anhydrous sodium sulfate, and after purification, an intermediate product B is obtained;

[0040] Preferably, in step ①, the mass concentration of the functionalized N-acetyl-D-glucosamine in the tert-butyl alcohol / water solution is 20-30 mg / mL;

[0041] Preferably, in step ①, in the tert-butyl alcohol / water solution, the mass ratio between the tert-butyl alcohol and the deionized water is 2-3:1;

[0042] Preferably, in step ①, the mass ratio between the functionalized N-acetyl-D-glucosamine, sodium ascorbate and copper sulfate pentahydrate is 10:1-3:2-3;

[0043] ② The intermediate product B prepared in step ① is dissolved in acetone, sodium methoxide in methanol is added, and after stirring at room temperature for 30-50 min, the pH is adjusted to neutral, and a solid is precipitated, which is filtered and collected, and after purification, an N-acetyl-D-glucosamine-containing composition is obtained;

[0044] Preferably, in step ②, the volume of the acetone is 10-20 mL;

[0045] Preferably, in step ②, the amount of sodium methoxide added is 10-16 mg.

[0046] The present application has the following beneficial effects:

[0047] The present application provides an anti-tumor N-acetyl-D-glucosamine-containing composition and a preparation method thereof. Through polypeptide modification and the application of N-acetyl-D-glucosamine, the immune regulation function of N-acetyl-D-glucosamine (GlcNAc) as a bioactive sugar molecule can have the following effects in tumor microenvironment regulation: sugar metabolism interference: cancer cells usually rely on sugar metabolism (Warburg effect) for rapid growth, and functional N-acetyl-D-glucosamine can inhibit the proliferation of cancer cells by affecting the sugar metabolism of cancer cells, and can synergistically play an anti-tumor role through sugar metabolism interference + immune regulation + apoptosis induction in three directions: N-acetyl-D-glucosamine affects the sugar metabolism and microenvironment of cancer cells, weakens the activity of cancer cells, and the polypeptide modifier enhances the penetration of drugs, and can promote the death of cancer cells by inhibiting tumor signaling pathways, and the combination of the two can improve the anti-cancer effect through immune enhancement and tumor targeting; hindering glycosylation: abnormal expression of glycosylated proteins on the surface of tumor cells can affect tumor metastasis and immune escape. N-acetyl-D-glucosamine can interfere with glycosylation modification and reduce the invasion ability of cancer cells; in the polypeptide modifier, oligoarginine can enhance the cell penetrating ability (CPP, Cell Penetrating Peptide), help the drug enter the cancer cells, and improve the treatment effect, oligoarginine can enhance cell oxidative stress and trigger the apoptosis pathway, causing the cancer cells to self-destruct, and the benzene sulfonamide part can reduce the survival signals of cancer cells by inhibiting protein tyrosine phosphatase (PTP), and benzene sulfonamide modification can bind to tumor-related proteins and improve drug targeting. BRIEF DESCRIPTION OF DRAWINGS

[0048] Figure 1 The figure is the nuclear magnetic resonance hydrogen spectrum image of the compound in Example 1 of the present application,

[0049] Figure 2 The figure is the inhibition activity result graph of the carbonic anhydrase of the composition prepared in Examples 1-3 and Comparative Examples 1-2 of the present application.

[0050] Figure 3 The figure is the inhibition activity result graph of HCT116 of the composition prepared in Examples 1-3 and Comparative Examples 1-2 of the present application.

[0051] The accompanying drawings are included to provide a further understanding of the present application, and constitute a part of the specification, and are used to explain the present application together with embodiments of the present application, and do not constitute a limitation of the present application. DETAILED DESCRIPTION

[0052] With reference to the drawings of the embodiments of the present application, the technical solutions in the embodiments of the present application will be described clearly and completely. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments of the present application; based on the embodiments of the present application, all the other embodiments obtained by a person of ordinary skill in the art without creative work fall within the scope of the present application.

[0053] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. Any methods and materials similar or equivalent to those described herein can be used in the practice of the present application. The described preferred methods and materials are only examples of the methods and materials that can be used in the practice of the present application.

[0054] The experimental methods in the following examples are all conventional methods unless otherwise specified; the experimental materials and experimental strains used in the following examples are all purchased from commercial channels unless otherwise specified.

[0055] Example 1

[0056] The present embodiment provides an anti-tumor N-acetyl-D-glucosamine-containing composition, which comprises the following components by weight: functionalized N-acetyl-D-glucosamine 1 part, polypeptide modifier 3 parts.

[0057] The preparation method of the functionalized N-acetyl-D-glucosamine specifically comprises the following steps:

[0058] S1, 1.2 g of D-glucosamine is dissolved in 30 mL of dichloromethane, placed in a 0°C environment, 1.5 mL of triethylamine is added, stirred and mixed for 10 min, 0.8 mL is mixed uniformly with 5 mL of dichloromethane to obtain a chloroformic acid-2,2,2-trichloroethyl ester / dichloromethane solution, the chloroformic acid-2,2,2-trichloroethyl ester / dichloromethane solution is slowly added dropwise, the reaction is kept at 0°C for 30-50 min, the reaction temperature is increased to room temperature, and the stirring reaction is continued for 2-3 h, the reaction pH is adjusted to neutral with saturated sodium bicarbonate solution, dichloromethane is added for extraction, the organic phase is washed with saturated sodium chloride aqueous solution, the organic phase is dried with anhydrous sodium sulfate, concentrated and purified to obtain N-Troc glucosamine;

[0059] S2, dissolve the N-Troc-amino glucose prepared in step S1 in 15 mL of dichloromethane, mix hydrobromic acid and acetic acid in a volume ratio of 1:2.5 under the condition of ice water bath, drop the hydrobromic acid-acetic acid solution, stir at a speed of 150 rpm, after 3 h of reaction, quench the reaction with deionized water, neutralize the reaction system to neutral with saturated sodium bicarbonate aqueous solution, extract with dichloromethane, collect the organic phase, wash with saturated sodium chloride aqueous solution, dry the organic phase with anhydrous sodium sulfate, concentrate and purify to obtain bromo-substituted-N-Troc-amino glucose;

[0060] S3, dissolve 0.5 g of bromo-substituted-N-Troc-amino glucose prepared in step S2 in 25 mL of DMF, add 0.3 g of sodium azide and mix uniformly, stir at a speed of 250 rpm at room temperature, after 45 min of reaction, quench the reaction with water, add ethyl acetate, collect the organic phase, wash with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain azido alpha-D-glucose;

[0061] S4, dissolve 1 g of azido alpha-D-glucose prepared in step S3 in 15 mL of anhydrous methanol, add 18 mL of glacial acetic acid, mix uniformly, add 0.2 g of zinc powder, and react at 60°C for 3 h, filter, collect the filtrate, add 20 mL of acetic anhydride, stir at a speed of 180 rpm, after 2 h of reaction, adjust the pH to neutral with saturated sodium bicarbonate aqueous solution, extract with ethyl acetate, wash with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain functionalized N-acetyl-D-amino glucose;

[0062] The method for preparing the polypeptide modifier specifically comprises the following steps:

[0063] A1, soak and swell the Rink Amide Resin resin in DCM for 30 min, remove the solvent, transfer the Rink Amide Resin resin to 15 vol% pyridine / DMF solvent, stir for 5 min, remove the pyridine / DMF solvent, add new pyridine / DMF solvent and continue to stir for 15 min, wash the Rink Amide Resin resin with DMF to obtain the deprotection group Rink Amide Resin resin;

[0064] A2. Place 1g of Fmoc-Pbf-arginine in a flask, add 20mL of DMF to fully dissolve Fmoc-Pbf-arginine, add 0.5g of HBTU and 0.2g of HOBt, mix at 150rpm, and after mixing evenly, add 0.4mL of DIEA, continue mixing for 5min, then add the Rink Amide Resin resin prepared in step A1, place at room temperature, stir at 200rpm, and react for 2h. After filtration, collect the solid Rink Amide Resin resin, wash the Rink Amide Resin resin repeatedly with DMF to obtain Rink Amide Resin resin-oligomeric arginine.

[0065] A3. Accurately weigh 0.8 g of p-carboxybenzenesulfonamide and place it in a flask. Add 20 mL of DMF to fully dissolve the p-carboxybenzenesulfonamide. Transfer the reaction system to an ice-water bath, add 3.2 mL of thionyl chloride, and stir at 300 rpm at room temperature. After stirring overnight, the reaction is detected by TLC to ensure complete reaction. Remove excess DMF by vacuum distillation to obtain intermediate product A. Add 30 mL of DMF to redissolve intermediate product A and mix it with Rink Amide Resin resin-oligoarginine from step A2. Add triethanolamine to adjust the pH of the reaction system to 8.5. React at 25 °C for 8 h. Extract with ethyl acetate, collect the organic phase, wash the organic phase with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain Rink Amide Resin resin-oligoarginine-benzenesulfonamide.

[0066] A4. Place the Rink Amide Resin-oligoarginine-benzenesulfonamide prepared in step A3 into 150 mL of LFA cutting solution, stir at 180 rpm for 2 h, filter, collect the filtrate, add pre-cooled anhydrous diethyl ether for precipitation, centrifuge, discard the supernatant, collect the precipitate, dry, and obtain oligoarginine-benzenesulfonamide.

[0067] A5. Accurately weigh 0.5 g of 4-pentynic acid and dissolve it in 10 mL of dichloromethane. Transfer the solution to an ice-water bath and add 3.5 mL of sulfoxide. Stir at 300 rpm for 12 h at room temperature. Add 1.5 g of potassium carbonate and mix thoroughly in an ice-water bath. Add the oligo-arginine-benzenesulfonamide prepared in step A4. Continue stirring and reacting at room temperature for 8 h. Extract with ethyl acetate, collect the organic phase, wash the organic phase with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain the peptide modified product.

[0068] The embodiment also provides a preparation method of the anti-tumor N-acetyl-D-glucosamine-containing composition, and specifically comprises the following steps:

[0069] ① 1g of functionalized N-acetyl-D-glucosamine and 3g of polypeptide modifier are dissolved in a t-butanol / water solution (volume ratio 2:1) according to weight parts, 0.1g of sodium ascorbate and 0.2g of copper sulfate pentahydrate are added, stirring is carried out at 80°C for 2h, then ethyl acetate is added for extraction, the organic phase is collected, dried with anhydrous sodium sulfate, and then purified to obtain an intermediate product B;

[0070] ② The intermediate product B prepared in step ① is dissolved in 10mL of acetone, 0.2mL of a 1ml / L methanolic sodium methoxide solution is taken, stirring is carried out at room temperature for 30min, then the pH is adjusted to neutral, solid is precipitated, filtration is carried out, the solid is collected, and then purification is carried out to obtain the N-acetyl-D-glucosamine-containing composition.

[0071] Figure 1 The nuclear magnetic resonance hydrogen spectrum image of the compound in Embodiment 1 of the application, wherein A is functionalized N-acetyl-D-glucosamine, B is a polypeptide modifier, and C is an N-acetyl-D-glucosamine-containing composition, as shown in the figure, A: 1 H-NMR (600 MHz, DMSO-d6) δ (ppm): 9.65 (s, 1H, -NH), 8.15 (d, 2H, -H), 7.65 (d, 2H, -H), 7.00 (br, 1H, NH), 5.25 (s, 1H, -CH), 3.85 (m, 2H, -CH2-NH-), 2.75 (t, 2H, -CH2-), 2.05 (t, 2H, -CH2-), 1.25 (s, 3H, -CH3).C: 1 H-NMR (600 MHz, DMSO-d6) δ (ppm): 9.65 (s, 1H, -NH), 8.15 (d, 2H, -H), 7.65 (d, 2H, -H), 7.00 (br, 1H, NH), 5.25 (s, 1H, -CH), 3.85 (m, 2H, -CH2-NH-), 2.75 (t, 2H, -CH2-), 2.05 (t, 2H, -CH2-), 1.25 (s, 3H, -CH3).C: 1H-NMR (600 MHz, DMSO-d6) δ (ppm): 9.65 (s, 1H, -NH), 8.15 (d, 2H), 7.65 (d, 2H), 7.00 (br, 1H, NH), 6.65 (s, 1H, -H), 5.25 (d, 1H, H-1), 4.90 (d, 1H, H-2), 3.85 (m, 1H, H-3), 3.25 (m, 2H, H-4, H-5), 2.75 (t, 2H, -CH2-), 2.05 (t, 2H, CH2-), 1.25 (s, 3H, -CH3).

[0072] Example 2

[0073] The embodiment provides an anti-tumor N-acetyl-D-glucosamine-containing composition, which comprises the following components in parts by weight: functionalized N-acetyl-D-glucosamine 2 parts, polypeptide modifier 3 parts.

[0074] The preparation method of the functionalized N-acetyl-D-glucosamine specifically comprises the following steps:

[0075] S1, 1 g of D-glucosamine is dissolved in 20 mL of dichloromethane, placed in a 0°C environment, 1.1 mL of triethylamine is added, stirred and mixed for 10 min, 1 mL is mixed with 5 mL of dichloromethane to obtain a chloroformic acid-2,2,2-trichloroethyl ester / dichloromethane solution, the chloroformic acid-2,2,2-trichloroethyl ester / dichloromethane solution is slowly added, the reaction is kept at 0°C for 30-50 min, the reaction temperature is increased to room temperature, and the stirring reaction is continued for 2-3 h, the reaction pH is adjusted to neutral with a saturated sodium bicarbonate solution, dichloromethane is added for extraction, the organic phase is washed with a saturated sodium chloride aqueous solution, the organic phase is dried with anhydrous sodium sulfate, and then concentrated and purified to obtain N-Troc glucosamine;

[0076] S2, 0.5 g of N-Troc-glucosamine prepared in step S1 is dissolved in 25 mL of dichloromethane, hydrogen bromide and acetic acid are mixed in a volume ratio of 1:2 under ice water bath conditions, the hydrogen bromide-acetic acid solution is added dropwise, stirring is carried out at a speed of 120 rpm, the reaction is carried out for 4 h, deionized water is added to quench the reaction, the reaction system is neutralized to neutral with a saturated sodium bicarbonate aqueous solution, dichloromethane is added for extraction, the organic phase is collected, washed with a saturated sodium chloride aqueous solution, dried with anhydrous sodium sulfate, and then concentrated and purified to obtain bromine-substituted-N-Troc-glucosamine;

[0077] S3, the bromo-substituted-N-Troc-amino glucose prepared in step S2 is dissolved in 50 mL of DMF, 0.2 g of sodium azide is added and mixed uniformly, stirring is carried out at room temperature at a speed of 200 rpm, after 60 min of reaction, water is added to quench the reaction, ethyl acetate is added, the organic phase is collected, washed with saturated sodium chloride solution, the organic phase is separated, dried with anhydrous sodium sulfate, filtered, the filtrate is collected, concentrated, purified, and azido alpha-D-glucose is obtained;

[0078] S4, 1 g of azido alpha-D-glucose prepared in step S3 is dissolved in 10 mL of anhydrous methanol, 10 mL of glacial acetic acid is added, mixed uniformly, 0.1 g of zinc powder is added, and the mixture is reacted at 50°C for 4 h, filtered, the filtrate is collected, 10 mL of acetic anhydride is added, stirring is carried out at a speed of 150 rpm, after 3 h of reaction, saturated sodium bicarbonate aqueous solution is added to adjust the pH to neutral, ethyl acetate is added for extraction, washed with saturated sodium chloride solution, the organic phase is separated, dried with anhydrous sodium sulfate, filtered, the filtrate is collected, concentrated, and purified, and functional N-acetyl-D-amino glucose is obtained;

[0079] The raw materials for preparing the polypeptide modifier include the following components in parts by weight: 4-aminobenzenesulfonamide 20 parts, oligoarginine 4, 5-hexynoic acid 30 parts;

[0080] The method for preparing the polypeptide modifier specifically includes the following steps:

[0081] A1, the Rink Amide Resin resin is soaked and swelled in DCM for 30 min, the solvent is removed, the Rink Amide Resin resin is transferred to 20 vol% pyridine / DMF solvent, stirring is carried out for 10 min, the pyridine / DMF solvent is removed, new pyridine / DMF solvent is added, stirring is continued for 10 min, the Rink Amide Resin resin is washed with DMF, and a deprotection group Rink Amide Resin resin is obtained;

[0082] A2, Fmoc-Pbf-arginine 1.2 g was accurately weighed into a flask, 40 mL of DMF was added to dissolve Fmoc-Pbf-arginine, HBTU 0.8 g, HOBt 0.16 g were added, mixed at 180 rpm, after mixing evenly, 0.6 mL of DIEA was added, and mixed for 5 min, then the Rink Amide Resin resin prepared in step A1 was added, stirred at 300 rpm at room temperature, reacted for 1 h, filtered, and the solid Rink Amide Resin resin was washed with DMF repeatedly to obtain Rink Amide Resin resin-oligarginine;

[0083] A3, accurately weigh p-carboxybenzenesulfonamide 1 g into a flask, add 20 mL of DMF to dissolve p-carboxybenzenesulfonamide, transfer the reaction system to an ice water bath, add 4.0 mL of dichlorosulfide, stir at room temperature at 200 rpm, stir overnight, after TLC detection, remove the excess DMF under reduced pressure to obtain intermediate A, add 30 mL of DMF to redissolve intermediate A, mix with Rink Amide Resin resin-oligarginine in step A2, add triethanolamine to adjust the pH of the reaction system to 9.0, react at 25°C for 8 h, then extract with ethyl acetate, wash the organic phase with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain Rink Amide Resin resin-oligarginine-benzenesulfonamide;

[0084] A4, the Rink Amide Resin resin-oligarginine-benzenesulfonamide prepared in step A3 was placed in 120 mL of TFA cleavage solution, stirred at 150 rpm for 3 h, filtered, the filtrate was collected, precipitated with pre-cooled anhydrous ether, centrifuged, the supernatant was discarded, the precipitate was collected and dried to obtain oligarginine-benzenesulfonamide;

[0085] A5, accurately weigh 4-pentynoic acid 0.4 g and dissolve in 10 mL of dichloromethane, transfer to an ice water bath, add dichlorosulfide 4.0 mL, stir at room temperature at 200 rpm for 16 h, then add potassium carbonate 1.6 g, mix evenly in an ice water bath, add oligarginine-benzenesulfonamide prepared in step A4, continue to stir at room temperature for 10 h, then extract with ethyl acetate, collect the organic phase, wash the organic phase with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain the polypeptide modifier;

[0086] The embodiment also provides a preparation method of the anti-tumor N-acetyl-D-glucosamine-containing composition, and specifically comprises the following steps:

[0087] ① 2g of functionalized N-acetyl-D-glucosamine and 4g of polypeptide modifier are dissolved in a t-butanol / water solution (volume ratio of 3:1) according to weight parts, 0.4g of sodium ascorbate and 0.6g of copper sulfate pentahydrate are added, and after stirring at 85℃ for 3h, ethyl acetate is added for extraction, the organic phase is collected, dried with anhydrous sodium sulfate, and after purification, an intermediate product B is obtained;

[0088] ② The intermediate product B prepared in step 1 is dissolved in 20mL of acetone, 0.25mL of 1ml / L methanolic sodium methoxide solution is taken, and after stirring at room temperature for 40min, the pH is adjusted to neutral, and a solid is precipitated, which is filtered and collected, and after purification, an N-acetyl-D-glucosamine-containing composition is obtained.

[0089] Embodiment 3

[0090] The embodiment provides an anti-tumor N-acetyl-D-glucosamine-containing composition, which comprises the following components according to weight parts: 3 parts of functionalized N-acetyl-D-glucosamine, 5 parts of polypeptide modifier.

[0091] The preparation method of the functionalized N-acetyl-D-glucosamine comprises the following steps:

[0092] S1, 1.2g of D-glucosamine is dissolved in 20mL of dichloromethane, and placed in a 0℃ environment, 1.2mL of triethylamine is added, and after stirring and mixing for 10min, 0.9mL of chloroformic acid-2,2,2-trichloroethyl ester / dichloromethane solution is obtained by mixing 5mL of dichloromethane, and the chloroformic acid-2,2,2-trichloroethyl ester / dichloromethane solution is slowly added dropwise, and the reaction is kept at 0℃ for 30-50min, and then the reaction temperature is increased to room temperature, and the stirring reaction is continued for 2-3h, the reaction pH is adjusted to neutral with saturated sodium bicarbonate solution, dichloromethane is added for extraction, the organic phase is washed with saturated sodium chloride aqueous solution, the organic phase is dried with anhydrous sodium sulfate, and then concentrated and purified to obtain N-Troc glucosamine;

[0093] S2, dissolve the N-Troc-amino glucose prepared in step S1 in 12.5 mL of dichloromethane, mix hydrobromic acid and acetic acid in a volume ratio of 1:3 under ice water bath conditions, add the hydrobromic acid-acetic acid solution dropwise, stir at a speed of 180 rpm, after 2 h of reaction, quench the reaction with deionized water, neutralize the reaction system to neutral with saturated sodium bicarbonate aqueous solution, extract with dichloromethane, collect the organic phase, wash with saturated sodium chloride aqueous solution, dry the organic phase with anhydrous sodium sulfate, concentrate and purify to obtain bromo-substituted-N-Troc-amino glucose;

[0094] S3, dissolve 0.5 g of bromo-substituted-N-Troc-amino glucose prepared in step S2 in 15 mL of DMF, add 0.4 g of sodium azide and mix uniformly, stir at a speed of 300 rpm at room temperature, after 30 min of reaction, quench the reaction with water, add ethyl acetate, collect the organic phase, wash with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain azido alpha-D-glucose;

[0095] S4, dissolve 1 g of azido alpha-D-glucose prepared in step S3 in 20 mL of anhydrous methanol, add 15 mL of glacial acetic acid, mix uniformly, add 0.3 g of zinc powder, and react at 70°C for 2 h, filter, collect the filtrate, add 15 mL of acetic anhydride, stir at a speed of 180 rpm, after 3 h of reaction, adjust the pH to neutral with saturated sodium bicarbonate aqueous solution, extract with ethyl acetate, wash with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain functionalized N-acetyl-D-amino glucose;

[0096] The method for preparing the polypeptide modifier specifically comprises the following steps:

[0097] A1, soak and swell the Rink Amide Resin resin in DCM for 30 min, remove the solvent, transfer the Rink Amide Resin resin to 25 vol% pyridine / DMF solvent, stir for 5 min, remove the pyridine / DMF solvent, add new pyridine / DMF solvent and continue to stir for 10 min, wash the Rink Amide Resin resin with DMF to obtain the deprotection group Rink Amide Resin resin;

[0098] A2, Fmoc-Pbf-arginine 1.2 g was placed in a flask, 30 mL of DMF was added to dissolve Fmoc-Pbf-arginine, HBTU 0.67 g, HOBt 0.4 g were added, mixed at 150 rpm, after mixing evenly, 0.4 mL of DIEA was added, and mixed for 5 min, then the Rink Amide Resin resin prepared in step A1 was added, stirred at 250 rpm at room temperature, reacted for 1.5 h, filtered, and the solid Rink Amide Resin resin was washed with DMF to obtain Rink Amide Resin resin-oligarginine;

[0099] A3, accurately weigh p-carboxybenzenesulfonamide 0.9 g into a flask, add 20 mL of DMF to dissolve p-carboxybenzenesulfonamide, transfer the reaction system to an ice water bath, add 4.5 mL of dichlorosulfide, stir at 250 rpm at room temperature, stir overnight, then detect the completion of the reaction by TLC, remove the excess DMF under reduced pressure to obtain intermediate A, dissolve intermediate A in 30 mL of DMF, mix with Rink Amide Resin resin-oligarginine in step A2, add triethanolamine to adjust the pH of the reaction system to 8.0, react at 25°C for 8 h, then extract with ethyl acetate, wash the organic phase with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain Rink Amide Resin resin-oligarginine-benzenesulfonamide;

[0100] A4, the Rink Amide Resin resin-oligarginine-benzenesulfonamide prepared in step A3 was placed in 100 mL of TFA cleavage solution, stirred at 150 rpm for 2.5 h, then filtered, collected the filtrate, precipitated with pre-cooled anhydrous ether, centrifuged, discarded the supernatant, collected the precipitate and dried to obtain oligarginine-benzenesulfonamide;

[0101] A5, accurately weigh 4-pentynoic acid 0.45 g and dissolve in 10 mL of dichloromethane, transfer to an ice water bath, add dichlorosulfide 3.0 mL, then stir at 250 rpm at room temperature for 14 h, add potassium carbonate 1.7 g, mix evenly in an ice water bath, then add oligarginine-benzenesulfonamide prepared in step A4, continue to stir at room temperature for 12 h, then extract with ethyl acetate, collect the organic phase, wash the organic phase with saturated sodium chloride solution, separate the organic phase, dry with anhydrous sodium sulfate, filter, collect the filtrate, concentrate and purify to obtain the polypeptide modifier.

[0102] The embodiment also provides a preparation method of the anti-tumor composition containing N-acetyl-D-glucosamine, and specifically comprises the following steps:

[0103] ① 3 g of functional N-acetyl-D-glucosamine and 5 g of polypeptide modifier are dissolved in a t-butyl alcohol / water solution (volume ratio of 2:1) according to weight parts, 0.9 g of sodium ascorbate and 0.9 g of copper sulfate pentahydrate are added, and after stirring at 90°C for 2 h, ethyl acetate is added for extraction, the organic phase is collected, and then dried with anhydrous sodium sulfate, to obtain an intermediate product B after purification;

[0104] ② The intermediate product B prepared in step ① is dissolved in 15 mL of acetone, 0.3 mL of a methanolic solution of sodium methoxide with a concentration of 1 ml / L is added, and after stirring at room temperature for 50 min, the pH is adjusted to neutral, and a solid is precipitated, which is collected by filtration, and then purified to obtain the composition containing N-acetyl-D-glucosamine.

[0105] Comparative Example 1

[0106] The comparative example provides an anti-tumor composition and a preparation method thereof, which are different from those of the embodiment 1 only in that steps A1-A4 are not included, and p-carboxybenzenesulfonamide is directly reacted with 4-pentynoic acid to obtain the modifier, and the remaining components and component contents are the same as those of the embodiment 1.

[0107] Comparative Example 2

[0108] The comparative example provides an anti-tumor composition and a preparation method thereof, which are different from those of the embodiment 1 only in that steps S2-S4 are not included, and the remaining components and component contents are the same as those of the embodiment 1.

[0109] Experimental Example 1

[0110] The carbonic anhydrase enzyme activity of the compositions prepared in Examples 1-3 and Comparative Examples 1-2 was determined. Carbonic anhydrase can catalyze the hydrolysis of 4-nitrophenyl acetate (4-NPA) to generate 4-nitrophenol. The change in absorbance of 4-nitrophenol at 405 nm was measured by a spectrophotometer to determine the enzyme activity of carbonic anhydrase. The inhibitory activity of the target compound on carbonic anhydrase was determined by a method that can only determine the inhibitory activity of the target compound on carbonic anhydrase. The method comprises the following steps: a buffer solution was prepared by dissolving 15 mmol / L 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, 0.01% tetraglycol dodecyl ether and 100 mmol / L sodium chloride, and the pH of the buffer solution was adjusted to 7.4. The carbonic anhydrase was diluted with the buffer solution to a concentration of 10 ng / μL. The 4-nitrophenyl acetate was diluted with the buffer solution to a substrate concentration of 1 nmol / L. The positive control acetazolamide was prepared by dissolving acetazolamide in DMSO to a concentration of 20 nmol / L. The target compound was prepared by dissolving the target compound in DMSO to a concentration of 10 nmo / L. 18 μL of the CAIX solution was taken and centrifuged. The acetazolamide and the target compound were diluted to different concentration gradients, respectively. The target compound was incubated with the enzyme at 25°C for 15 min to form an enzyme-inhibitor complex. Then, the substrate 4-NPA was added to the complex solution, and the solution was incubated at 25°C for 90 min. The absorbance was measured, and the IC50 of the compound on carbonic anhydrase was calculated by a standard curve.

[0111] Figure 2 The results of the inhibitory activity of the compositions prepared in Examples 1-3 and Comparative Examples 1-2 on carbonic anhydrase are shown in the following figure. The IC50 of the compositions prepared in Examples 1-3 is less than 200 mmol / L, the IC50 of the composition prepared in Comparative Example 1 is 540.5 mmol / L, and the IC50 of the composition prepared in Comparative Example 2 is 708.6 mmol / L. The above data show that the compositions prepared in Examples 1-3 have a significant inhibitory activity on carbonic anhydrase. The oligo-arginine and the sulfonamide group with inhibitory activity are modified on the outside of the NK cell. The arginine has more guanidino groups, which can interact with the hydrophobic part of the active site, thereby producing an inhibitory effect on carbonic anhydrase.

[0112] Experimental Example 2

[0113] The cell activity of the compositions prepared in Examples 1-3 and Comparative Examples 1-2 was determined. 5-fluorouracil (5-FU) was used as a positive control drug, and the MTT method was used to test the in vitro anti-proliferative activity of the target compound on human colon cancer cells (HCT116). The cells in logarithmic growth were trypsinized, and the cell suspension was adjusted to a concentration of about 5×10 4The cells were added to a 96-well plate at 100 μL per well, and a blank control was added with only culture solution, and was placed in a 37°C, 5% CO2 incubator for 24 h. The culture solution of the experimental group was replaced with a new culture solution containing different concentrations of the sample to be tested, and the culture solution of the control group was replaced with a culture solution containing an equal volume of solvent, and each group had 3 parallel holes, and was incubated at 37°C, 5% CO2 for 48 h. The supernatant was discarded, and the cells were washed twice with PBS, and 100 μL of MTT solution (5 mg·mL-1) was added to each well, and was incubated for 4 h. The supernatant was discarded, 100 μL of dimethyl sulfoxide was added to each well, and the 96-well plate was placed on a micro-vibration instrument for 3 min, and was mixed, and then the optical density (OD) value at 492 nm was measured using an enzyme-labeled instrument. The cell viability inhibition rate was calculated according to the negative control group, and the IC50 value of each compound was obtained by fitting the curve;

[0114]

[0115] wherein, : the average OD value of the blank control group; : the average OD value of the test group.

[0116] Figure 3 The inhibition activity of the compositions prepared in Examples 1-3 and Comparative Examples 1-2 of the present application on HCT116 is shown in the graph, and the IC50 of Examples 1-3 is less than 70 μmol / L, while the IC50 of the composition prepared in Comparative Example 1 is 234.5 μmol / L, and the IC50 of the composition prepared in Comparative Example 2 is 486.6 μmol / L, which shows that the compositions prepared in Examples 1-3 have obvious inhibition activity on carbonic anhydrase. The oligoguanidine and the sulfonyl group with inhibitory activity are modified on the outside of the NK cell, and the nitrogen atoms of the sulfonamide group form coordination with Zn 2+ The sulfonamide group also forms hydrogen bond interactions with Thr200 and Thr201, respectively, and the oxygen atoms on the N-acetyl-D-glucosamine sugar ring form hydrogen bond interactions with Thr73, and the guanidino group of the oligoguanidine forms hydrophobic interactions with Gln92, so the compositions of the comparative examples cannot interact with the active center amino acids, thereby reducing the activity.

[0117] Although the embodiments of the present application have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present application.

[0118] The above describes the present application and its embodiments, which are not limited, and the drawings only show one of the embodiments of the present application, and the actual application is not limited thereto. In general, if a person skilled in the art is inspired by it, without departing from the purpose of the present application, without creative design, similar ways and embodiments of the technical solution should belong to the protection scope of the present application.

Claims

1. An anti-colon cancer N-acetyl-D-glucosamine containing compound, characterized in that: The compound is prepared by reaction of a functionalized N-acetyl-D-glucosamine and a polypeptide modifier; The preparation method of the functionalized N-acetyl-D-glucosamine specifically comprises the following steps: S1, taking D-glucosamine is dissolved in dichloromethane, placed in 0 DEG C environment, adding triethylamine, stirring mixed 10 min, slowly drop 2,2,2-trichloroethyl chloroformate / dichloromethane solution, keep 0 DEG C condition, reaction 30-50 min, increase reaction temperature to room temperature, continue stirring reaction 2-3 h, using saturated sodium bicarbonate solution to adjust the reaction pH to neutral, adding dichloromethane for extraction, with saturated sodium chloride aqueous solution washes organic phase, with anhydrous sodium sulfate drying organic phase, concentrated, purified, N-Troc glucosamine is obtained; S2, the N-Troc-glucosamine prepared in step S1 is dissolved in dichloromethane, drop 2,2,2-trichloroethyl chloroformate / dichloromethane solution in ice water bath conditions, stirring is carried out according to 120-180 rpm, after reaction 2-4 h, adding deionized water for quenching reaction, with saturated sodium bicarbonate aqueous solution neutralization reaction system to neutral, adding dichloromethane for extraction, collection organic phase, with saturated sodium chloride aqueous solution for washing, with anhydrous sodium sulfate drying organic phase, concentrated, purified, bromo-substituted-N-Troc-glucosamine is obtained; S3, the bromo-substituted-N-Troc-glucosamine prepared in step S2 is dissolved in DMF, adding sodium azide for mixing uniform, stirring is carried out at room temperature according to 200-300 rpm, after reaction 30-60 min, adding water for quenching reaction, adding ethyl acetate, collection organic phase, with saturated sodium chloride solution for washing, separation organic phase is dried with anhydrous sodium sulfate, filtration, collection filtrate, concentrated, purified, azido alpha-D-glucose is obtained; S4, the azido alpha-D-glucose prepared in step S3 is dissolved in anhydrous methanol, adding glacial acetic acid, mixing uniform, adding zinc powder, placed in 50-70 DEG C for 2-4 h, filtration, collection filtrate, transferred to ice water bath conditions, adding acetic anhydride, stirring reaction is carried out according to 150-180 rpm, after reaction 2-3 h, adding saturated sodium bicarbonate aqueous solution to adjust pH to neutral, adding ethyl acetate for extraction, with saturated sodium chloride solution for washing, separation organic phase is dried with anhydrous sodium sulfate, filtration, collection filtrate, concentrated, purified, functionalized N-acetyl-D-glucosamine is obtained; The preparation method of the polypeptide modifier specifically comprises the following steps: A1, after the Rink Amide Resin resin is placed in DCM for soaking and swelling, the solvent is removed, the Rink Amide Resin resin is transferred to a pyridine / DMF solution, after stirring for 5-10 min, the pyridine / DMF solvent is removed, new pyridine / DMF solvent is added, and stirring is continued for 10-15 min, then the Rink Amide Resin resin is washed with DMF to obtain a deprotection group Rink Amide Resin resin; A2, Fmoc-Pbf-arginine is dissolved in DMF, HBTU and HOBt are added, mixed thoroughly, DIEA is added, stirring is continued at a speed of 150-180 rpm for 3-5 min, the deprotection group Rink Amide Resin resin prepared in step A1 is added, stirring is carried out at room temperature at a speed of 200-300 rpm for 1-2 h, then filtration is performed, the Rink Amide Resin resin is collected, and repeatedly washed with DMF to obtain a Rink Amide Resin resin-oligomeric arginine; A3, p-carboxybenzenesulfonamide is dissolved in DMF, dichlorosulfoxide is added dropwise under an ice water bath, stirring is carried out at room temperature at a speed of 200-300 rpm overnight, then TLC is used to detect the completion of the reaction, the solvent is removed to obtain an intermediate product A, the intermediate product A is dissolved in DMF, and added to the reaction system in step A2, TEA is added to keep the pH of the reaction system at 8-9, the reaction is carried out at 25-40℃ for 4-8 h, then ethyl acetate is added for extraction, the organic phase is collected, washed with saturated sodium chloride solution, separated, dried with anhydrous sodium sulfate, filtered, the filtrate is collected, concentrated, and purified to obtain a Rink Amide Resin resin-oligomeric arginine-benzenesulfonamide; A4, the Rink Amide Resin resin-oligomeric arginine-benzenesulfonamide prepared in step A3 is placed in a TFA cleavage solution, stirring is carried out at a speed of 150-180 rpm for 2-3 h, then filtration is performed, the filtrate is collected, pre-cooled anhydrous ether is added for precipitation, centrifugation is performed, the supernatant is discarded, the precipitate is collected and dried to obtain an oligomeric arginine-intermediate product A; A5, 4-pentynoic acid is dissolved in dichloromethane, placed in an ice water bath, dichlorosulfoxide is added, the temperature is increased to room temperature, stirring is carried out at a speed of 200-300 rpm for 12-16 h, then potassium carbonate is added, transferred to ice water, and added to the oligomeric arginine-benzenesulfonamide prepared in step A4, stirring is carried out at room temperature at a speed of 200-300 rpm for 8-12 h, then ethyl acetate is added for extraction, the organic phase is collected, washed with saturated sodium chloride solution, separated, dried with anhydrous sodium sulfate, filtered, the filtrate is collected, concentrated, and purified to obtain a polypeptide modifier; The preparation method of the anti-colon cancer N-acetyl-D-glucosamine-containing compound specifically comprises the following steps: In step S1, the weight concentration of the D-glucosamine in dichloromethane is 40-60 mg / mL; The mass-volume ratio between the D-glucosamine and triethylamine is 0.7-0.9 g / mL; 2. The N-acetyl-D-glucosamine containing compound against colon cancer according to claim 1, characterized in that: The mass-volume ratio between the D-glucosamine and chloroformic acid-2,2,2-trichloroethyl ester is 1-1.2 g / mL. In step S2, the mass concentration of the N-Troc-glucosamine in dichloromethane is 20-40 mg / mL; The volume ratio between the hydrobromic acid and acetic acid in the hydrobromic acid-acetic acid solution is 1:2-3; 3. The N-acetyl-D-glucosamine containing compound against colon cancer according to claim 2, characterized in that: The mass-volume ratio between the N-Troc-glucosamine and the hydrobromic acid-acetic acid solution is 0.05-0.15 g / mL. In step S3, the mass concentration of the bromo-substituted-N-Troc-glucosamine in DMF is 10-30 mg / mL; The mass ratio between the bromo-substituted-N-Troc-glucosamine and sodium azide is 5:2-4.

4. The N-acetyl-D-glucosamine containing compound against colon cancer according to claim 3, characterized in that: In step S4, the mass concentration of the azido α-D-glucose in anhydrous methanol is 0.05-0.1 g / mL; The volume ratio between the glacial acetic acid and anhydrous methanol is 0.75-1.2:1; 5. A colon cancer containing N-acetyl-D-glucosamine compound according to claim 4, characterized by: The added mass of zinc is 10-30% of the mass of the azido α-D-glucose; The mass-volume ratio between the azido α-D-glucose and acetic anhydride is 10-50 mg / mL. In step A1, the volume fraction of pyridine in DMF in the pyridine / DMF solution is 15%-25%; In step A2, the mass concentration of the Fmoc-Pbf-arginine in DMF is 30-50 mg / mL; 6. The N-acetyl-D-glucosamine containing compound against colon cancer according to claim 5, characterized in that: In step A2, the mass ratio between the Fmoc-Pbf-arginine, HBTU and HOBt is 1.5-2:1:0.2-0.6; In step A2, the mass-volume ratio between the Fmoc-Pbf-arginine and DIEA is 2.0-2.5 g / mL In step A3, the mass concentration of the p-carboxybenzenesulfonamide in DMF is 40-50 mg / mL; In step A3, the mass-volume ratio between the p-carboxybenzenesulfonamide and dichlorosulfoxide is 0.2-0.25 g / mL; ​ ​ In step A4, the volume of the TFA cleavage solution is 100-150 mL; In step A5, the mass concentration of the 4-pentynoic acid in dichloromethane is 40-50 mg / mL; In step A5, the mass-volume ratio between the 4-pentynoic acid and dichlorosulfoxide is 0.1-0.15 g / mL: In step A5, the mass ratio between the 4-pentynoic acid and potassium carbonate is 1:2.5-4.

7. A colon cancer containing N-acetyl-D-glucosamine compound according to claim 6, characterized by: In step ①, the mass ratio between the t-butyl alcohol and deionized water in the t-butyl alcohol / water solution is 2-3:1; In step ①, the mass ratio between the available N-acetyl-D-glucosamine, sodium ascorbate, and copper sulfate pentahydrate is 10:1-3:2-3; In step ②, the volume of the acetone is 10-20 mL; In step ②, the amount of sodium methoxide added is 10-16 mg.

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