Sulfated derivatives of fucosylated chondroitin sulfate oligosaccharides and uses thereof

Abstract

The application provides sulfated derivatives of fucosylated chondroitin sulfate oligosaccharides and application thereof, and belongs to the technical field of marine drugs. The structure of the sulfated derivatives of fucosylated chondroitin sulfate oligosaccharides is mainly a disaccharide repeating unit with GlcA and GalNAc connected through a beta-1,3 / beta-1,4 glycosidic bond as a main chain, and a fucose connected to the O-3 position of GlcA as a side chain; sulfation occurs at any 1-3 positions of C4 and / or C6 of the acetylglucosamine in the main chain, and C2,3,4 of the fucose in the side chain. Through in-vivo and in-vitro level verification, the application plays an anti-tumor metastasis role by significantly inhibiting the adhesion of tumors to endothelium, platelets and the like and tumor angiogenesis, so as to use S-dFCS as an active substance, prepare a medicine or a health product with the aid of a pharmaceutically acceptable carrier, and be capable of being used for preventing and treating tumors and diseases related to postoperative metastasis.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of marine organisms and medicine, and particularly relates to a sulfated derivative of fucosylated chondroitin sulfate oligosaccharide and application thereof. BACKGROUND

[0002] Tumor is a common and serious disease endangering human health in the world, and its incidence rate is only next to cardiovascular disease. Although the level of chemotherapy, radiotherapy and surgical treatment of tumor has been greatly improved compared with the past, due to the easy metastasis of malignant tumor, it brings great difficulty to clinical medication, surgical treatment and the like. On the one hand, the proliferation of tumor cells is inhibited by chemical drugs, which often has great side effects, and the effect on circulating tumor cells with low proliferation rate is poor, and it is difficult to control the metastasis and recurrence of tumor; on the other hand, postoperative metastasis of tumor is the main reason for treatment failure and death of most cancer patients, and such postoperative metastasis is often systemic and organ metastasis, which cannot be controlled by drug chemotherapy. Tumor metastasis often leads to serious consequences such as infection, cachexia, hemorrhage, immune disorder, organ damage, metabolic abnormality and the like, finally causes the deterioration of the disease, and even death. Therefore, it is of great significance to develop a safe and effective drug for preventing postoperative recurrence and metastasis of tumor.

[0003] Fucosylated chondroitin sulfate (FCS) is a glycosaminoglycan (GAG) derived from the body wall of sea cucumber, which is a natural acidic mucopolysaccharide with fucose branch, and the sulfation degree of GalNAc and Fuc residues is high. FCS has a variety of potential biological activities, but most of the current researches are still focused on its anticoagulant and antithrombotic effects. In recent years, FCS has also attracted many researchers to explore its antitumor activity, and Chinese patents CN 110776578B, CN 106349397A and CN 101724086B report that the depolymerization product of FCS has activities such as anti-inflammatory and anti-vascular lesion on the basis of significantly reduced bleeding tendency.

[0004] However, the current research on the antitumor activity of FCS is still insufficient, and no drug application is carried out, and it still has great research potential. SUMMARY

[0005] The application aims to provide a sulfated derivative of fucosylated chondroitin sulfate oligosaccharide, and provide specific application of the derivative, thereby expanding the use of the series of compounds in the prevention and treatment of postoperative metastasis of tumor.

[0006] To achieve the above application purposes, the application adopts the following technical solutions:

[0007] A sulfated derivative (S-dFCS) of fucosylated chondroitin sulfate oligosaccharide, the structure of which is mainly a backbone of disaccharide repeating units of GlcA and GalNAc connected by β-1,3 / β-1,4 glycosidic bonds, with a side chain of fucose connected to the O-3 position of GlcA; sulfated at any 1-3 of the C4 and / or C6 positions of the acetylglucosamine in the backbone and the C2,3,4 positions of the fucose in the side chain.

[0008] Further, the S-dFCS has the structural formula (I) as shown:

[0009]

[0010] wherein n = 0-4; R1, R2, R3 = -H or -SO3H.

[0011] Further, the sulfated derivative of fucosylated chondroitin sulfate oligosaccharide has a degree of polymerization less than 21 and a degree of sulfation of 40%-60%; the S-dFCS is a sulfated derivative of fucosylated chondroitin sulfate oligosaccharide with a weight average molecular weight of 500 Da to 20,000 Da.

[0012] Further, the preparation method of the S-dFCS is:

[0013] (1) obtaining an intermediate product by deacetylation of fucosylated chondroitin sulfate polysaccharide extracted from the body wall and / or internal organs of animals in the class Holothuroidea of the phylum Echinozoa;

[0014] (2) obtaining fucosylated chondroitin sulfate oligosaccharide (dFCS) by nitrite degradation;

[0015] (3) obtaining a sulfated derivative of fucosylated chondroitin sulfate oligosaccharide (S-dFCS) by sulfation reaction with a sulfation reagent.

[0016] Further, the sulfation reagent is any one or a mixture of more than one of sulfur trioxide·triethylamine, sulfur trioxide·pyridine or chlorosulfonic acid·pyridine.

[0017] The S-dFCS is used in the preparation of a drug or health product for preventing or treating tumors.

[0018] The pharmaceutically acceptable salt of the sulfated derivative of fucosylated chondroitin sulfate oligosaccharide is used in the preparation of a drug or health product for preventing or treating tumors; the pharmaceutically acceptable salt is a sodium salt, a potassium salt or a calcium salt.

[0019] The types of tumors include, but are not limited to, common cancer types such as breast cancer, lung cancer, melanoma, colon cancer or gastric cancer.

[0020] The application mechanism is that the sulfated derivatives of FCS oligosaccharide interact with adhesion molecules P, L-selectin and vascular endothelial growth factor VEGF, thereby inhibiting the adhesion of tumor cells to endothelial cells, platelets and the like and tumor angiogenesis; the S-dFCS oligosaccharide inhibits tumor cell migration, thereby resisting tumor metastasis and having no toxic side effects, that is, the S-dFCS oligosaccharide plays a role in inhibiting the spread and metastasis of tumor cells and is highly efficient and non-toxic; the S-dFCS oligosaccharide inhibits chicken embryo chorioallantoic membrane (CAM) angiogenesis, thereby resisting tumor metastasis, that is, the S-dFCS oligosaccharide plays a role in inhibiting the abnormal neovascularization of tumor-related blood vessels and destroying the tumor metastasis microenvironment.

[0021] The application is specifically for the development of a tumor metastasis prevention and / or treatment drug or health care product for circulating tumor cells and their metastasis microenvironment; in addition, the application is also suitable for postoperative patients and sub-healthy people after tumor chemotherapy, radiotherapy and surgical treatment.

[0022] A drug for resisting tumor, which comprises one or more combinations of sulfated derivatives of fucosylated chondroitin sulfate hexaose, sulfated derivatives of fucosylated chondroitin sulfate nonaose, sulfated derivatives of fucosylated chondroitin sulfate dodecaose, sulfated derivatives of fucosylated chondroitin sulfate pentadecaose and sulfated derivatives of fucosylated chondroitin sulfate octadecaose.

[0023] A health care product for resisting tumor, which comprises one or more combinations of sulfated derivatives of fucosylated chondroitin sulfate hexaose, sulfated derivatives of fucosylated chondroitin sulfate nonaose, sulfated derivatives of fucosylated chondroitin sulfate dodecaose, sulfated derivatives of fucosylated chondroitin sulfate pentadecaose and sulfated derivatives of fucosylated chondroitin sulfate octadecaose.

[0024] The drug is in the form of tablets, capsules, oral liquids, injections, powders, ointments or external liquid medicines, supplemented with various pharmaceutically acceptable carriers; or is used in combination with other anti-tumor drugs; the health care product is in the form of tablets, capsules, oral liquids, powders, ointments and the like, supplemented with other edible carriers.

[0025] Further, the sulfated derivatives of fucosylated chondroitin sulfate oligosaccharide (S-dFCS) are subjected to in-vitro anti-tumor research: the S-dFCS can intervene in the action of various cytokines and adhesion molecules in the tumor microenvironment, significantly inhibit tumor cell migration, spread and abnormal neovascularization, thereby destroying the tumor metastasis microenvironment and having an obvious dose-effect relationship; at the same time, the S-dFCS has no tumor cell toxicity, which is different from chemotherapeutic drugs and is a highly efficient and low-toxic anti-tumor metastasis drug.

[0026] Further, the sulfated derivative (S-dFCS) of the fucosylated chondroitin sulfate oligosaccharide is subjected to in-vivo anti-tumor research: the experimental lung metastasis model of mice is established by injecting the high-metastasis cell B16F10 of melanoma into the tail vein of the mice, and the S-dFCS can significantly inhibit the experimental lung metastasis of the mice, and when the dosage is 15 mg / kg, the lung metastasis of the tumor is almost completely inhibited, and the alveolar structure of the lung tissue is restored to be complete, and the effect is significantly better than that of the unmodified oligosaccharide (dFCS). Therefore, the S-dFCS provided by the application expands the application of the series of compounds in the prevention and treatment of postoperative metastasis of tumors, and improves the in-vivo and in-vitro activity of the series of compounds in inhibiting tumor metastasis.

[0027] The advantages and beneficial effects of the application are as follows:

[0028] The sulfated derivative of the fucosylated chondroitin sulfate oligosaccharide provided by the application has many advantages such as rich source, simple preparation process, relatively uniform structure, mild safety, high stability and the like.

[0029] It is verified in practice that the S-dFCS has a significant effect of inhibiting tumor metastasis at in-vivo and in-vitro levels, and the S-dFCS is used as an active substance to prepare a medicine by using a pharmaceutically acceptable carrier, and is used for preventing and treating tumors and diseases related to postoperative metastasis of tumors. In addition, the S-dFCS can also be used in combination with clinically used cancer treatment drugs (chemotherapy and targeted drugs) to improve the treatment effect of tumors. BRIEF DESCRIPTION OF DRAWINGS

[0030] Figure 1 It is the fluorescence integral graph of the combination of S-dFCS and cytokines (P, L-selectin, VEGF); wherein A is the combination with P-selectin; B is the combination with L-selectin; and C is the combination with VEGF.

[0031] Figure 2 It is the influence of S-dFCS on the proliferation of melanoma cells (B16F10).

[0032] Figure 3 It is the influence of S-dFCS on the migration of melanoma cells (B16F10); wherein A is a photographing image under a microscope; and B is a quantitative column chart.

[0033] Figure 4 It is the influence of S-dFCS on the growth of blood vessels of chicken embryo allantoic membrane (CAM).

[0034] Figure 5 It is the influence of S-dFCS on the lung nodules of melanoma (B16F10) metastasis mice; wherein A is a representative image of lung nodules of the experimental mice; and B is a quantitative column chart of lung nodules of the experimental mice.

[0035] Figure 6 Effect of S-dFCS on lung pathological structure of melanoma (B16F10) metastasis mice; A is the NC group; B is the MD group; C is the dFCS group; D is the low-dose S-dFCS group; E is the high-dose S-dFCS group. DETAILED DESCRIPTION

[0036] Other advantages and features of the present application will become more apparent from the following detailed description of the application when read in conjunction with the accompanying drawings. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. The following examples are given to illustrate the application and should not be construed to limit the scope of the application. The examples are presented for the purpose of clarity and a person of ordinary skill in the art will be able to employ the teachings of the application without undue experimentation.

[0037] The fucosylated chondroitin sulfate (FCS) is obtained by defatting, enzymatic hydrolysis, CPC complex precipitation, alcohol precipitation, dialysis and strong anion exchange column separation. The fucosylated chondroitin sulfate oligosaccharide (dFCS) is obtained by using nitrous acid degradation method with selective glycosidic bond. The oligosaccharide derivative S-dFCS is obtained by sulfation modification of dFCS. The sulfation degree of S-dFCS is 40% to 60%, which is higher than that of dFCS. The effects of dFCS and S-dFCS on anti-tumor metastasis are evaluated by molecular level, cell level and in vivo animal experiment.

[0038] Example 1 Preparation of fucosylated chondroitin sulfate oligosaccharide (dFCS)

[0039] FCS 1.0 g is weighed in a reaction bottle, dissolved in 25 mL of hydrazine hydrate, then 250 mg of hydrazine sulfate is added, and N 2 The reaction is stirred at 90°C for 30 h under protection. After cooling to room temperature, four times the volume of anhydrous ethanol is added to the reaction solution, and alcohol precipitation is repeated several times. The precipitate is redissolved and dialyzed (molecular weight cut-off 1000 Da). The dialysate is concentrated and freeze-dried to obtain the deacetylated product DaFCS sample. 100 mg of DaFCS is weighed and dissolved in 5 mL of H2O at 0°C, then 10 mL of 5.5M nitrous acid solution is added, and the reaction is stirred at 0°C for 3 h. After the reaction is completed, 1.0M NaOH is added to adjust the pH to 8.0-9.0 to terminate the reaction. 300 mg / mL NaBH4 0.6 mL is added to the reaction solution, and the reaction is heated at 50°C for 2 h. After cooling to room temperature, 0.5M H2SO4 is added to adjust the pH to about 4.0 to remove excess NaBH4. Finally, 1.0M NaOH is added to adjust the pH to about 7.0, dialyzed, concentrated, and freeze-dried to obtain the FCS oligosaccharide (dFCS).

[0040] The structural formula of the dFCS prepared in this example is shown in formula (II), wherein n = 0-4; R1, R2 = H or -SO3H; the degradation degree is detected by HPGPC method, and the FCS oligosaccharide mixture of hexose, nonose, dodecaose, pentadecaose and octadecaose with uniform structure can be obtained by the method of this example, and the sulfation degree is between 20% and 40%.

[0041]

[0042] Example 2 Preparation of sulfated derivatives (S-dFCS) of FCS oligosaccharide

[0043] 200 mg of dFCS was weighed and dissolved in 20 mL of anhydrous DMF, and after dissolution, 2.0 g of sulfur trioxide triethylamine was added. Under N2 atmosphere, the reaction was heated and stirred at 70°C for 24 h. After the reaction was completed, the reaction bottle was placed in an ice bath, and after the reaction liquid was cooled to about 0°C, saturated sodium bicarbonate solution was slowly added to adjust pH to about 8.0. After 30 min of continuous reaction, dialysis and freeze-drying were performed to obtain the sulfated derivatives (S-dFCS) of FCS oligosaccharide.

[0044] The structural formula of the sulfated FCS oligosaccharide prepared in this example is shown in formula (I) above. The sulfation degree of the product after modification under this condition is between 40% and 60%, which indicates that the -OH of Fuc has not been completely sulfated, which also avoids the bleeding side effects that may be caused by over-sulfation.

[0045]

[0046] wherein n = 0-4; R1, R2, R3 = H or -SO3H.

[0047] Example 3 Determination of molecular weight of FCS oligosaccharide and its sulfated derivatives

[0048] The weight average molecular weight of the sample was determined by high performance gel permeation chromatography (HPGPC) combined with multi-angle laser light scattering (MALLS). The chromatographic conditions are as follows: chromatographic column: Shodex Ohpak SB-803HQ (8.0 μm x 300 mm) and Shodex Ohpak SB-802.5HQ (8.0 μm x 300 mm) chromatographic column in series; mobile phase: 0.1 M Na2SO4 solution; detector: differential detector combined with multi-angle laser light scattering detector. The data were processed by Astra software to calculate the molecular weight of the sample.

[0049] Example 4 Binding of S-dFCS to P, L-selectin and VEGF

[0050] Glass substrate was soaked in Piranha solution for 30 min, washed with water for three times, and dried. Then the substrate was soaked in APTMS ethanol solution (10%) for 30 min, washed with ethanol. The substrate was soaked in dichloromethane solution containing BIBB (1%) and TEA (1%), washed with dichloromethane and ethanol, and dried. 2 Dried. HEMA (2.86 M), BPY (60.7 mM) and CuBr (24.6 mM) were dissolved in a mixed solution of methanol and water with equal proportion, and degassed. N 2 Dried. The polymerization reaction was completed in the protected reactor. The substrate was washed with ethanol and water, and dried. 2 Dried. The chip was soaked in a solution of CC and DIPEA in acetone, and reacted at 4°C for 8 h, washed with acetone, and dried. 2 Dried. Microarrays were fabricated on the substrate surface by spotting, and the immobilization of saccharide compounds on the substrate surface was achieved by incubation overnight in a humidified chamber. The chip was blocked with EOA (1 M, pH 8.60) as needed. His-FITC was dissolved or diluted in Tris-HCl buffer solution (25 mM, pH 7.60; containing 1 mM CaCl2, 1 mM MnCl2); P-selection-His tag, L-selection-His tag, and VEGF-His tag were dissolved or diluted in PBS (10 mM, pH 7.40). The above obtained solutions were incubated with the saccharide chip, and the binding signal was detected by a fluorescence chip scanner (excitation wavelength 488 nm).

[0051] The results are shown in Figure 1 S-dFCS can bind to P, L-selection, and VEGF; the binding strength of S-dFCS to P-selectin is higher than 1000, the binding strength of S-dFCS to L-selectin and VEGF is higher than 2000, and both are higher than the binding ability of unmodified oligosaccharide.

[0052] The results preliminarily show that S-dFCS can inhibit the process of tumor metastasis by inhibiting the adhesion of tumor cells to endothelial cells and platelets and the mechanism of tumor angiogenesis.

[0053] Example 5 Effect of S-dFCS on the proliferation of melanoma cells

[0054] B16F10 cells in the logarithmic growth phase were digested with 0.25% (w / v) trypsin containing 0.02% (w / v) EDTA, and 100 μL of 3×10 4Cell suspensions of cells / mL were incubated at 37°C for 24 h. Cells were then treated with a series of sample concentrations, with six replicates per concentration, and incubated for another 24 h. Afterward, 20 μL of LCK-8 solution was added to each well, and the cells were incubated at 37°C in the dark for 4 h. The absorbance (OD value) at 450 nm was then measured using a microplate reader.

[0055] The results are as follows Figure 2 As shown, dFCS, S-dFCS and their purified nonaglycone had no effect on the proliferation of B16F10 cells within the detected concentration range (25–400 μg / mL), indicating that they had no toxic effect on tumor cells within this range and were highly safe, making them suitable for subsequent experimental studies.

[0056] Example 6: Effect of S-dFCS on melanoma cell migration

[0057] B16F10 cells in the logarithmic growth phase were harvested and starved for 12 hours in FBS-free medium. The starved cells were then resuspended in FBS-free medium and diluted to a density of 5 × 10⁻⁶ cells / mL. 5 / mL. Simultaneously, add 100μL of FBS-free culture medium to the Transwell chamber for activation for 15 min. Add 100μL of cell suspension to the upper chamber and add complete culture medium containing 20% ​​FBS to the lower chamber. Treat the cells in the upper chamber with different concentrations of samples, with three replicates for each experiment. Incubate the 24-well plate in an incubator for 24 h. Then, gently wipe the inner wall of the chamber with a clean cotton swab, discard the culture medium in the wells, fix with 4% paraformaldehyde at room temperature for 20 min, stain with 0.1% crystal violet at room temperature for 20 min, rinse the chamber with PBS 2-3 times, allow it to air dry, observe and photograph it under a microscope, and destain with 33% acetic acid. Measure the absorbance (OD value) at 570 nm using a microplate reader.

[0058] The results are as follows Figure 3 As shown, after 24 hours of incubation in the untreated Control group, a large number of tumor cells migrated to the submembrane of the Transwell chamber, indicating that B16F10 cells themselves have strong migration ability. Treatment with different concentrations of S-dFCS significantly inhibited the migration ability of melanoma cells in a concentration-dependent manner. At a concentration of 100 μg / mL, S-dFCS had a significant inhibitory effect on melanoma cells, and at the same concentration, its inhibitory effect was superior to the positive control (LMWH).

[0059] Example 7: Effect of S-dFCS on angiogenesis in the chorioallantoic membrane of chicken embryos

[0060] Tumor angiogenesis is a complex process, and hypoxia, inflammation, and external mechanical stimulation are all contributing factors. Inhibiting tumor angiogenesis is a hot topic in anti-tumor research, and it holds promise for the development of effective and safe drugs and health products that inhibit tumor growth and metastasis.

[0061] Chicken embryos were pre-incubated for 7 days. Observation was performed using an egg candler, and the location of the embryonic head, air cell size, and ventilation point were marked with a pencil. After alcohol sterilization in a laminar flow hood, a small hole was made in the outer shell of the air cell using curved ophthalmic scissors. A small opening of approximately 10mm × 10mm was then carefully made using tweezers, gently breaking the shell membrane around the opening to separate the allantoic membrane from the shell membrane. A pre-prepared sterile mixed cellulose filter membrane (5mm in diameter) was placed on the allantoic membrane in a relatively avascular area. 20μL of sample solutions of different concentrations were added to the sterile mixed cellulose filter membrane using a micropipette. The opening was then sealed with sterile medical tape, and the embryo was further incubated. After 48 hours, the tape was removed, and changes in blood vessels were observed. The adjacent areas of newly formed blood vessels were photographed using a digital camera.

[0062] The results are as follows Figure 4 As shown, the Control group treated with saline exhibited vigorous angiogenesis and increased vascular density, with a radially growing vascular network visible around the drug application point. In contrast, the high and low dose S-dFCS groups showed a significant reduction in the number and distribution of blood vessels, with a stronger effect than the unmodified oligosaccharide dFCS group. Furthermore, the inhibitory effect on CAM angiogenesis tended to increase with increasing S-dFCS concentration, further confirming that the drug's anti-metastatic effect is related to its inhibition of angiogenesis.

[0063] Example 8: Effect of S-dFCS on experimental lung metastases of melanoma in mice

[0064] C57BL / 6J mice were randomly divided into five groups (n=12). B16F10 cells were diluted with 0.9% NaCl (MD group), dFCS (15 mg / kg), S-dFCS (5 mg / kg), and S-dFCS (15 mg / kg), respectively. After pre-incubation for 30 min, each mouse was injected intravenously with 1.5 × 10⁻⁶ cells. 5 Tumor cells were identified. The mice were then administered the drug every two days, with their body weight monitored. After 23 days, the mice were dissected, their lungs removed, fixed in formalin, and examined for lung metastasis. Paraffin sections of the lung tissue were prepared and stained with hematoxylin and eosin (HE). The average number of metastatic nodules in each group was calculated using the presence of tumor metastases in the lungs as an indicator.

[0065] The results are as follows Figure 5It is shown that, compared with the model group, FCS oligosaccharide (dFCS) and sulfated derivative of FCS oligosaccharide (S-dFCS) can significantly reduce lung metastasis of melanoma in mice, and the sulfated modification of FCS oligosaccharide can enhance the inhibition of lung metastasis of tumor; and the high-dose S-dFCS group (15 mg / kg) shows a more superior anti-tumor effect than the low-dose (5 mg / kg) group, and the lung of the mouse in the high-dose S-dFCS group has no obvious nodules, and the lung metastasis is almost completely inhibited; as shown in Figure 6 As shown in the figure, under the microscope after HE staining, it can be observed that the lung nuclei of the model group are large and the cells are arranged closely; and the lung tissue structure of the sample group is relatively complete.

[0066] In summary, through in vivo and in vitro experiments, the sulfated derivative (S-dFCS) of fucosylated chondroitin sulfate oligosaccharide can intervene in various cytokines and adhesion molecules in the tumor microenvironment, thereby playing a role in inhibiting tumor cell migration, invasion and inhibiting tumor angiogenesis, and its good effect is better than that of the unmodified oligosaccharide compound dFCS, and the sulfated modification of depolymerized FCS has a higher potential application value in preventing and treating tumor metastasis.

[0067] The present application innovatively finds that fucosylated chondroitin sulfate oligosaccharide has good activity in inhibiting tumor metastasis after sulfated modification; S-dFCS has many advantages such as rich source, simple preparation process, relatively uniform structure, mild safety, high stability, etc., and can be used for developing health care products or drugs for preventing or treating tumor metastasis in the tumor metastasis microenvironment, and has a broad development and application prospect.

[0068] Obviously, the above embodiments of the present application are only examples for clearly illustrating the present application, and the protection scope of the present application is not limited thereto; any skilled person in the art can make other different forms of changes or variations without departing from the spirit and technical scope of the present application; here, all the embodiments cannot be exhausted, and any obvious changes or variations derived from the technical solutions of the present application still fall within the protection scope of the present application.

[0069] On the basis of the above embodiments, the technical features involved therein and the functions and effects of the technical features in the present application are described in detail to help the skilled person in the art to fully understand the technical solutions of the present application and to reproduce them.

[0070] Finally, although the present specification is described in terms of embodiments, not every embodiment exhibits every characteristic or implements every combination of features described in the present specification. The present specification has been described in a manner that is thorough and complete to one skilled in the art and the specification is intended to be construed as an exemplification of one or more embodiments rather than as an exhaustive list of embodiments.

Claims

1. A sulfated derivative of a fucosylated chondroitin sulfate oligosaccharide, characterized in that, The structure of the derivative is a backbone of disaccharide repeat units of GlcA and GalNAc connected by β-1,3 / β-1,4 glycosidic bonds, and a side chain of fucose connected to the O-3 position of GlcA; any 1-3 positions of C4 and / or C6 of the acetylglucosamine in the backbone and C2,3,4 of the fucose in the side chain are sulfated; the degree of polymerization of the derivative is less than 21, the degree of sulfation is 40%-60%, and the weight average molecular weight is between 500 Da and 20000 Da; the structural formula of the derivative is as follows: ； wherein n=0-4; R1, R2, R3=-H or -SO3H.

2. Process for the preparation of sulfated derivatives of the fucosylated chondroitin sulfate oligosaccharides according to claim 1, characterized in that, The method comprises the following steps: (1) obtaining an intermediate product by deacetylation of fucosylated chondroitin sulfate polysaccharide extracted from the body wall and / or internal organs of animals in the class Holothuroidea of the phylum Echinozoa; (2) obtaining fucosylated chondroitin sulfate oligosaccharide dFCS by nitrite degradation; (3) obtaining sulfated derivative S-dFCS of fucosylated chondroitin sulfate oligosaccharide by sulfation reaction of the fucosylated chondroitin sulfate oligosaccharide dFCS with a sulfation reagent.

3. Use of the sulfated derivative of fucosylated chondroitin sulfate oligosaccharide according to claim 1 in the preparation of a medicament for preventing or treating melanoma.

4. Use of a pharmaceutically acceptable salt of the sulfated derivative of fucosylated chondroitin sulfate oligosaccharide according to claim 1 in the preparation of a medicament for preventing or treating melanoma.

5. A medicament for use against melanoma, characterized in that, The medicament comprises the sulfated derivative of fucosylated chondroitin sulfate oligosaccharide according to claim 1.

Images