CN117510679A - Sulfated derivative of fucosylated chondroitin sulfate oligosaccharide and application thereof - Google Patents
Sulfated derivative of fucosylated chondroitin sulfate oligosaccharide and application thereof Download PDFInfo
- Publication number
- CN117510679A CN117510679A CN202311469593.1A CN202311469593A CN117510679A CN 117510679 A CN117510679 A CN 117510679A CN 202311469593 A CN202311469593 A CN 202311469593A CN 117510679 A CN117510679 A CN 117510679A
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- CN
- China
- Prior art keywords
- chondroitin sulfate
- fucosylated chondroitin
- derivative
- sulfated derivative
- sulfated
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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Classifications
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- C08B37/0069—Chondroitin-4-sulfate, i.e. chondroitin sulfate A; Dermatan sulfate, i.e. chondroitin sulfate B or beta-heparin; Chondroitin-6-sulfate, i.e. chondroitin sulfate C; Derivatives thereof
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- A—HUMAN NECESSITIES
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- A61P35/04—Antineoplastic agents specific for metastasis
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The invention provides a sulfated derivative of fucosylated chondroitin sulfate oligosaccharide and application thereof, belonging to the technical field of marine medicaments. The structure of the sulfated derivative of the fucosylated chondroitin sulfate oligosaccharide is characterized in that disaccharide repeated units of GlcA and GalNAc which are connected through beta-1, 3/beta-1, 4 glycosidic bonds are taken as a main chain, and fucose connected to the GlcA O-3 position is taken as a side chain; sulfation occurs at the C4 and/or C6 positions of the backbone acetamido galactose, at any 1-3 of the C2,3,4 positions of the branched fucose. Through in vivo and in vitro level verification, the invention plays a role in resisting tumor metastasis by remarkably inhibiting the adhesion of tumors to endothelium, blood platelets and the like and tumor angiogenesis, takes S-dFCS as an active substance and is matched with a pharmaceutically acceptable carrier to prepare medicines or health care products, and can be used for preventing and treating tumors and postoperative metastasis related diseases thereof.
Description
Technical Field
The invention belongs to the technical field of marine biological medicines, and particularly relates to a sulfated derivative of fucosylated chondroitin sulfate oligosaccharide and application thereof.
Background
Tumors are one of the most common and serious diseases endangering human health in today's world, with incidence of disease being inferior to cardiovascular disease. Although the current chemotherapy, radiotherapy and operation treatment level of tumor are greatly developed and improved compared with the past, the malignant tumor is easy to be transferred, and great difficulty is brought to clinical medication, operation treatment and the like. On one hand, the chemical drugs are used for inhibiting the proliferation of tumor cells, so that the side effect is large, the effect on circulating tumor cells with low proliferation rate is poor, and the metastasis and recurrence of tumors are difficult to control; on the other hand, tumor postoperative metastasis is a major cause of failure and death in most cancer patients, and often, the postoperative metastasis is systemic multi-organ metastasis and cannot be controlled by drug chemotherapy. Tumor metastasis often results in serious consequences such as infection, cachexia, hemorrhage, immune malfunction, organ damage, metabolic abnormalities, etc., ultimately leading to worsening of the condition and even death. Therefore, the development of a safe and effective medicament for preventing the recurrence and metastasis after tumor operation has very important significance.
Fucosylated chondroitin sulfate (Fucosylated chondroitin sulfate, FCS), a Glycosaminoglycan (GAG) derived from the body wall of sea cucumber, is a natural acidic Glycosaminoglycan with fucose branches, and has a high degree of sulfation on GalNAc and Fuc residues. FCS has a variety of potential biological activities, but most of the research is still focused on its anticoagulant and antithrombotic effects. In recent years, FCS has attracted research on antitumor activity by several researchers, and chinese patent CN 110776578B, CN 106349397A, CN 101724086B reports that FCS depolymerized products have anti-inflammatory and anti-vasculopathy activities on the basis of significantly reduced bleeding tendency.
However, current research on FCS antitumor activity is still insufficient, nor is it applied as a drug, which still has great research potential.
Disclosure of Invention
The invention aims to provide a sulfated derivative of fucosylated chondroitin sulfate oligosaccharide and a specific application of the derivative, so that the application of the series of compounds in prevention and treatment of tumor postoperative metastasis is expanded.
In order to achieve the aim of the invention, the invention is realized by adopting the following technical scheme:
a sulfated derivative (S-dFCS) of a fucosylated chondroitin sulfate oligosaccharide, the derivative having a structure in which a disaccharide repeating unit formed by connecting GlcA and GalNAc via a β -1,3/β -1,4 glycosidic bond is used as a main chain, and fucose attached to the GlcA O-3 position is used as a side chain; sulfation occurs at the C4 and/or C6 positions of the backbone acetamido galactose, at any 1-3 of the C2,3,4 positions of the branched fucose.
Further, the structural formula of the S-dFCS is shown as (I):
wherein n=0 to 4; r1, R2, R3 = -H or-SO 3 H。
Further, the degree of polymerization of the sulfated derivative of the fucosylated chondroitin sulfate oligosaccharide is less than 21, and the degree of sulfation is 40% -60%; the S-dFCS is a sulfated derivative of a fucosylated chondroitin sulfate oligosaccharide having a weight average molecular weight of 500Da to 20000 Da.
Further, the preparation method of the S-dFCS comprises the following steps:
(1) Deacetylating fucoidan polysaccharide extracted from body wall and/or viscera of Holothuria nobilis of Echinodermata to obtain intermediate product;
(2) Degrading nitrous acid to obtain fucosylated chondroitin sulfate oligosaccharide (dFCS);
(3) The sulfated derivative of fucosylated chondroitin sulfate oligosaccharide (S-dFCS) is obtained by subjecting it to a sulfation reaction by a sulfating agent.
Further, the sulfating agent is any one or more of sulfur trioxide-triethylamine, sulfur trioxide-pyridine or chlorosulfonic acid-pyridine.
The application of the S-dFCS in preparing medicaments or health-care products for preventing or treating tumors.
The application of the pharmaceutically acceptable salt of the sulfated derivative of the fucosylated chondroitin sulfate oligosaccharide in preparing medicaments or health-care products for preventing or treating tumors; the pharmaceutically acceptable salt is sodium salt, potassium salt or calcium salt.
The tumor types include, but are not limited to, common cancer species such as breast cancer, lung cancer, melanoma, colon cancer or stomach cancer.
The mechanism of the application is as follows: sulfated derivatives of FCS oligosaccharides inhibit adhesion of tumor cells to endothelial cells, platelets, etc. and tumor angiogenesis by interacting with adhesion molecules P, L-selectin, vascular endothelial growth factor VEGF; the S-dFCS oligosaccharide inhibits tumor cell migration so as to resist tumor metastasis, has no toxic or side effect, namely plays a role in inhibiting tumor cell diffusion and metastasis, and is efficient and nontoxic; the chick embryo chorioallantoic membrane (CAM) angiogenesis is inhibited to resist tumor metastasis, namely the chick embryo chorioallantoic membrane (CAM) angiogenesis is inhibited, and the chick embryo chorioallantoic membrane has the effects of inhibiting abnormal angiogenesis related to tumor and destroying the tumor metastasis microenvironment.
The application is specifically as follows: for developing a tumor metastasis preventing and/or treating drug or health care product aiming at circulating tumor cells and a metastasis microenvironment thereof; in addition, the composition is also suitable for postoperative patients and sub-health people after tumor chemotherapy, radiotherapy and operation treatment.
A medicament for use in anti-tumour comprising one or more combinations of a sulfated derivative of fucosylated chondroitin sulfate hexasaccharide, a sulfated derivative of fucosylated chondroitin sulfate nonasaccharide, a sulfated derivative of fucosylated chondroitin sulfate dodecasaccharide, a sulfated derivative of fucosylated chondroitin sulfate pentasaccharide, a sulfated derivative of fucosylated chondroitin sulfate octadeca saccharide.
A health product for resisting tumor comprises one or more of sulfated derivative of fucosylated chondroitin sulfate hexasaccharide, sulfated derivative of fucosylated chondroitin sulfate nine saccharide, sulfated derivative of fucosylated chondroitin sulfate dodecasaccharide, sulfated derivative of fucosylated chondroitin sulfate pentasaccharide, and sulfated derivative of fucosylated chondroitin sulfate octadecanose.
The medicine is in the form of tablets, capsules, oral liquid, injection, powder, paste or external liquid medicine, and is supplemented with various pharmaceutically acceptable carriers; or is compounded with other antitumor drugs; the health care product is in the form of tablets, capsules, oral liquid, powder, ointment and the like, and is supplemented with other edible carriers.
Further, in vitro antitumor studies were performed on the sulfated derivative of fucosylated chondroitin sulfate oligosaccharide (S-dFCS): S-dFCS can interfere with the actions of various cytokines and adhesion molecules in tumor microenvironment, obviously inhibit the migration, diffusion and abnormal angiogenesis of tumor cells, thereby destroying the tumor metastasis microenvironment and having obvious dose-effect relationship; meanwhile, the S-dFCS has no tumor cytotoxicity, is different from a chemotherapeutic drug, and is an efficient low-toxicity anti-tumor metastasis drug.
Further, in vivo antitumor study was performed on the sulfated derivative of fucosylated chondroitin sulfate oligosaccharide (S-dFCS): the experimental lung metastasis model is established by using the high metastasis cell B16F10 of the melanoma of the mice through tail vein injection, S-dFCS can obviously inhibit experimental lung metastasis of the melanoma of the mice, and when the dosage is 15mg/kg, the lung metastasis of the tumor is almost completely inhibited, the pulmonary alveolar structure of the lung tissue is restored to be complete, and the effect is obviously better than that of unmodified oligosaccharide (dFCS). Therefore, the S-dFCS provided by the invention expands the application of the series of compounds in the prevention and treatment of tumor postoperative metastasis and improves the activity of the series of compounds in inhibiting tumor metastasis in vivo and in vitro.
The invention has the advantages and beneficial effects that:
the sulfated derivative of the fucosylated chondroitin sulfate oligosaccharide provided by the invention has the advantages of rich sources, simple preparation process, uniform structure, mildness, safety, high stability and the like.
The practical verification proves that the S-dFCS has the effect of obviously inhibiting tumor metastasis at both in vivo and in vitro levels, and the S-dFCS is taken as an active substance and is used for preparing medicines by being assisted with pharmaceutically acceptable carriers, so that the S-dFCS is used for preventing and treating tumors and postoperative metastasis related diseases thereof. In addition, the S-dFCS can be combined with clinically used medicines (chemotherapy and targeted medicines) for treating cancers, so that the treatment effect of tumors is improved.
Drawings
FIG. 1 is a graph showing the fluorescence integration of the binding between S-dFCS and cytokine (P, L-selectin, VEGF); wherein A is binding to P-selectin; b is the binding to L-selectin; c is binding to VEGF.
FIG. 2 shows the effect of S-dFCS on proliferation of melanoma cells (B16F 10).
FIG. 3 is the effect of S-dFCS on migration of melanoma cells (B16F 10); wherein A is a photographed image under a microscope; b is a quantitative histogram.
FIG. 4 is the effect of S-dFCS on chick embryo allantoic membrane (CAM) vascular growth.
FIG. 5 is the effect of S-dFCS on pulmonary nodules in melanoma (B16F 10) metastatic mice; wherein A is a representative image of a pulmonary nodule of an experimental mouse; b is a quantitative histogram of pulmonary nodules of experimental mice.
FIG. 6 is the effect of S-dFCS on the lung pathological tissue structure of melanoma (B16F 10) metastatic mice; a is NC group; b is MD group; c is dFCS group; d is a low dose S-dFCS group; e is the high dose S-dFCS group.
Detailed Description
Other advantages and features of the present invention will become apparent upon reading the following detailed description of the invention in conjunction with the drawings. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art. Reagents and materials used in the following examples are commercially available unless otherwise specified.
The invention obtains the Fucosylated Chondroitin Sulfate (FCS) through degreasing, enzymolysis, CPC complex precipitation, alcohol precipitation, dialysis and strong anion exchange column separation. The method for degrading FCS by nitrous acid with glycosidic bond selectivity is adopted to degrade the FCS to obtain fucosylated chondroitin sulfate oligosaccharide (dFCS), the fucosylated chondroitin sulfate oligosaccharide is taken as a raw material to be sulfated and modified to obtain an oligosaccharide derivative S-dFCS, and the sulfation degree of the S-dFCS is 40% -60% and is higher than that of dFCS. And then evaluating the effect of dFCS and S-dFCS on resisting tumor metastasis through molecular level, cell level and in vivo animal experiment tests.
EXAMPLE 1 preparation of fucosylated chondroitin sulfate oligosaccharide (dFCS)
After weighing FCS1.0g in a reaction flask and dissolving in 25mL of hydrazine hydrate, 250mg of hydrazine sulfate, N 2 The reaction was stirred at 90℃for 30h under protection. Cooling to room temperature, adding four times of absolute ethyl alcohol into the reaction solution, repeating alcohol precipitation for multiple times, redissolving the precipitate, dialyzing (with a molecular weight cut-off of 1000 Da), concentrating the dialysate, and freeze-drying to obtain a deacetylation product DaFCS sample. 100mg of DaFCS was weighed and dissolved in 5mL of H at 0 ℃ 2 After O, 10mL of 5.5M nitrous acid solution is added, the mixture is stirred at 0 ℃ for 3 hours, and after the reaction is finished, 1.0M NaOH is added for regulating the pH value to 8.0-9.0, and the reaction is stopped. 300mg/mL NaBH was added to the reaction solution 4 0.6mL was heated at 50deg.C for 2h, cooled to room temperature, and then 0.5MH was added 2 SO 4 Adjusting pH to about 4.0 removes excess NaBH 4 Finally, regulating the pH to be approximately 7.0 by using 1.0MNaOH, dialyzing, concentrating, and freeze-drying to obtain the FCS oligosaccharide (dFCS).
The structural formula of dFCS prepared in the embodiment is shown as a formula (II), wherein n=0 to 4; r1, r2=h or-SO 3 H is formed; the degradation degree is detected by HPGPC, and the FCS oligosaccharide mixture of hexasaccharide, nonasaccharide, dodecasaccharide, pentasaccharide and octadecane with uniform structure can be obtained by the method, and the sulfation degree is measured to be between 20% and 40%.
EXAMPLE 2 preparation of sulfated derivatives of FCS oligosaccharides (S-dFCS)
200mg of dFCS was dissolved in 20mL of anhydrous DMF, and 2.0g of thiotriethylamine trioxide, N was added after dissolution 2 And (3) heating and stirring at 70 ℃ for reaction for 24 hours under the atmosphere, placing a reaction bottle in an ice bath after the reaction is finished, slowly adding saturated sodium bicarbonate solution after the reaction liquid is cooled to about 0 ℃, adjusting the pH to about 8.0, continuing to react for 30 minutes, and dialyzing and freeze-drying to obtain the sulfated derivative (S-dFCS) of the FCS oligosaccharide.
The structural formula of the sulfated FCS oligosaccharide prepared in the embodiment is shown as the formula (I), and the sulfation degree of the product after modification under the condition is between 40% and 60%, which indicates that the-OH of Fuc is not completely sulfated, and meanwhile, the bleeding side effect possibly caused by oversulfate is avoided.
Wherein n=0 to 4; r1, R2, r3=h or-SO 3 H。
EXAMPLE 3 molecular weight determination of FCS oligosaccharides and sulfated derivatives thereof
The weight average molecular weight of the samples was determined using High Performance Gel Permeation Chromatography (HPGPC) in combination with an eighteen angle laser light scattering (MALLS). The chromatographic conditions were as follows: chromatographic column: shodex Ohpak SB-803HQ (8.0 μm. Times.300 mm) and Shodex Ohpak SB-802.5HQ (8.0 μm. Times.300 mm) columns were connected in series; mobile phase: 0.1M Na 2 SO 4 A solution; a detector: the differential detector is used in combination with an eighteen angle laser scatterometer detector. The molecular weight of the samples was calculated by data processing with Astra software.
EXAMPLE 4 binding of S-dFCS to P, L-selection, VEGF
The glass substrate is soaked in Piranha washing liquid for 30min, washed with water for three times, dried, soaked in APTMS ethanol solution (10%) for reaction for 30min, and washed with ethanol. Soaking the substrate in dichloromethane solution containing BIBB (1%) and TEA (1%) for 30min, washing with dichloromethane and ethanol, and N 2 And (5) blow-drying. HEMA (2.86M), BPY (60.7 mM) and CuBr (24.6 mM) were dissolved in a mixed solution of methanol and water in equal proportions, and pre-desorptedAnd (3) air. Adding N 2 In the protected reactor, the polymerization reaction is finished, the substrate is washed by ethanol and water, N 2 And (5) blow-drying. Soaking the chip in acetone solution of CC and DIPEA, reacting at 4deg.C for 8 hr, washing with acetone, and N 2 And (5) blow-drying. Microarrays were prepared on the substrate surface by spotting, the immobilization of saccharide compounds on the substrate surface was achieved by incubating overnight with moisture, and the chip was properly blocked with EOA (1 m, ph 8.60). His-FITC was buffered in Tris-HCl (25 mM, pH 7.60; containing 1mM CaCl) 2 、1mM MnCl 2 ) Dissolving or diluting; the P-selection-His tag, L-selection-His tag, VEGF-His tag were dissolved or diluted with PBS (10 mM, pH 7.40). Each of the solutions obtained above was incubated with a sugar chip, and the binding signal was detected by a fluorescence chip scanner (excitation wavelength 488 nm).
As a result, as shown in FIG. 1, S-dFCS can bind to P, L-selection, VEGF; wherein, the binding strength of S-dFCS and P-selectin is higher than 1000, the binding strength of S-dFCS and P-selectin is higher than 2000, and the binding capacity of S-dFCS and P-selectin is higher than that of L-selectin, VEGF.
The results preliminarily indicate that S-dFCS can inhibit tumor metastasis process by inhibiting the mechanism of tumor cell adhesion to endothelium and platelets and tumor angiogenesis.
Example 5 effect of S-dFCS on proliferation of melanoma cells
Taking B16F10 cells in logarithmic phase, digesting with 0.25% (w/v) pancreatin containing 0.02% (w/v) EDTA, mixing thoroughly, adding 100 μl of 3×10 per well into 96-well plate 4 The cell suspension was cultured at 37℃for 24 hours. Sample treatment cells of a series of concentrations were added, each concentration was plated with 6 duplicate wells, and culture was continued for 24h. Thereafter, 20. Mu. LCCK-8 solution was added to each well, incubated at 37℃for 4 hours in the absence of light, and absorbance at 450nm (OD value) was measured by an ELISA reader.
As shown in FIG. 2, dFCS, S-dFCS and purified nonasaccharide thereof have no influence on proliferation of B16F10 cells in the concentration range (25-400 mu g/mL), namely have no toxic effect on tumor cells in the range, and have high safety, and can be used for subsequent experimental study.
Example 6 effect of S-dFCS on migration of melanoma cells
Taking B16F10 cells in logarithmic growth phase, starving with FBS-free medium for 12h, re-suspending the starved cells in FBS-free medium, and diluting the cell density to 5×10 5 /mL. Simultaneously, 100. Mu.L of FBS-free medium was added to the Transwell chamber and activated for 15min. 100. Mu.L of the cell suspension was added to the upper chamber and the complete medium containing 20% FBS was added to the lower chamber. The upper chamber cells were treated with samples of different concentrations, 3 replicates were set for each set of experiments, and 24 well plates were placed in an incubator for 24h. Then, the inner wall of the cell was gently wiped with a clean cotton swab, kong Zhongpei solution was discarded, the cell was fixed with 4% paraformaldehyde at room temperature for 20min, stained with 0.1% crystal violet at room temperature for 20min, rinsed with PBS for 2 to 3 times, naturally air-dried, then placed under a microscope for observation and photographing, and after decolorization with 33% acetic acid, absorbance (OD value) at 570nm was detected by an enzyme-labeled instrument.
As shown in fig. 3, it can be seen that after the Control group which is not treated by the cells is incubated for 24 hours, a great amount of tumor cells migrate to the position under the membrane of the Transwell chamber, which indicates that the migration capacity of the B16F10 cells is stronger; after being treated by S-dFCS with different concentrations, the migration capacity of melanoma cells is obviously inhibited, and the melanoma cells have concentration dependence. At a concentration of 100 μg/mL, S-dFCS has a significant inhibition of melanoma cells, and at the same concentration, it is superior to the positive control (LMWH).
EXAMPLE 7 Effect of S-dFCS on chick embryo chorioallantoic vascular growth
Angiogenesis of tumors is a complex process, and hypoxia, inflammation, external mechanical stimulus and the like are all causative factors of angiogenesis of tumors. Inhibition of tumor angiogenesis is a hotspot problem in anti-tumor research, and is expected to be developed into effective and safe medicaments and health-care products for inhibiting tumor growth and metastasis.
The chick embryo was pre-incubated for 7 days, and the embryo head position, air chamber size and fenestration were marked with a pencil, as observed with an egg candler. After alcohol disinfection in an ultra-clean workbench, a small hole is knocked on the shell of the chick embryo air chamber by using elbow ophthalmic scissors, a small hole with the diameter of about 10mm multiplied by 10mm is peeled off by using forceps, and the eggshell membrane around the small hole is carefully clipped and broken, so that the allantoic membrane and the shell membrane are separated. A sterile mixed cellulose filter membrane chip (diameter: 5 mm) prepared in advance was placed in a relatively avascular region on the allantoic membrane, 20. Mu.L of sample solutions of different concentrations were dropped onto the sterile mixed cellulose filter membrane chip with a micropipette, then sealed with a sterile medical adhesive tape, and embryos were further incubated. After 48 hours, the tape was peeled off, the vessel was observed for changes and the adjacent neonatal vessel area was photographed with a digital camera.
As a result, as shown in FIG. 4, the blood vessel growth of the Control group to which physiological saline was administered was vigorous, and the blood vessel density was increased, so that a vascular network radially grown around the dosing point was seen; the number and distribution of blood vessels under the action of the S-dFCS high and low dose group are obviously reduced, and the effect is stronger than that of the unmodified oligosaccharide dFCS group. Meanwhile, with the increase of the concentration of S-dFCS, the inhibiting effect of CAM angiogenesis tends to be enhanced, and the anti-metastasis effect of the drug is further proved to be related to the inhibiting effect of angiogenesis.
Example 8 effect of S-dFCS on experimental lung metastasis in melanoma in mice
C57BL/6J mice were randomly divided into five groups (n=12), and B16F10 cells were diluted with 0.9% NaCl (MD group), dFCS (15 mg/kg), S-dFCS (5 mg/kg), S-dFCS (15 mg/kg), respectively, and each tail vein of each mouse was injected 1.5×10 after pre-incubation for 30min 5 And (3) tumor cells. Subsequently, the mice were dosed once every two days while their body weights were monitored. After 23 days, the mice were dissected, the lungs were removed, fixed in formalin, and examined for lung metastasis. And paraffin sections were performed on lung tissue, and HE staining was performed on lung tissue. The average node number of each group was calculated using tumor lung metastasis nodes as an indicator.
As shown in fig. 5, both FCS oligosaccharide (dFCS) and sulfated derivative of FCS oligosaccharide (S-dFCS) significantly reduced pulmonary metastasis in murine melanoma compared to the model group, and sulfation modification of FCS oligosaccharide enhanced tumor pulmonary metastasis inhibition; and the high-dose S-dFCS group (15 mg/kg) shows more excellent anti-tumor effect than the low-dose (5 mg/kg), and the high-dose S-dFCS group mice have no obvious nodules in the lung and almost completely inhibit lung metastasis; as shown in fig. 6, when the model group is observed under HE staining and then under a mirror, the model group has large lung nuclei and compact cell arrangement; while the lung tissue structure of the sample group was restored to be relatively intact.
In conclusion, the in-vitro and in-vivo experiments prove that the sulfated derivative (S-dFCS) of the fucosylated chondroitin sulfate oligosaccharide plays roles in inhibiting tumor cell migration, invasion and tumor angiogenesis by interfering with various cytokines and adhesion molecules in a tumor microenvironment, has good effects superior to those of an unmodified oligosaccharide compound dFCS, carries out sulfation modification on depolymerized FCS, and has potential application values for more effectively preventing and treating tumor metastasis.
The invention innovatively discovers that the fucosylated chondroitin sulfate oligosaccharide has good activity of inhibiting tumor metastasis after being sulfated and modified; the S-dFCS has the advantages of rich sources, simple preparation process, uniform structure, mildness, safety, high stability and the like, can be used for developing health care products or medicaments for preventing or treating tumor metastasis aiming at tumor metastasis microenvironment, and has wide development and application prospects.
It is obvious that the above-described embodiments of the present invention are only examples for clearly illustrating the present invention, and the scope of the present invention is not limited thereto; other variations or modifications of the present invention may be apparent to those of ordinary skill in the art, without departing from the spirit and scope of the present invention; it is not intended to be exhaustive of all embodiments, and all obvious variations or modifications that come within the scope of the invention are within the scope of the invention.
The present invention has been described in detail with reference to the above embodiments, and the functions and actions of the features in the present invention will be described in order to help those skilled in the art to fully understand the technical solution of the present invention and reproduce it.
Finally, although the description has been described in terms of embodiments, not every embodiment is intended to include only a single embodiment, and such description is for clarity only, as one skilled in the art will recognize that the embodiments of the disclosure may be combined as appropriate to form other embodiments that will be apparent to those skilled in the art.
Claims (7)
1. A sulfated derivative of fucosylated chondroitin sulfate oligosaccharide, which is characterized in that the structure of the derivative takes disaccharide repeated units of GlcA and GalNAc connected through beta-1, 3/beta-1, 4 glycosidic bonds as a main chain and takes fucose connected at the O-3 position of GlcA as a side chain; sulfation occurs at the C4 and/or C6 positions of the main chain acetamido galactose, at any 1-3 of the C2,3,4 positions of the branched fucose; the structural formula of the derivative is shown as follows:
；
wherein n=0 to 4; r1, R2, R3 = -H or-SO 3 H。
2. The sulfated derivative of a fucosylated chondroitin sulfate oligosaccharide as defined in claim 1, characterized in that the derivative has a degree of polymerization of less than 21, a degree of sulfation of 40% -60%, and a weight average molecular weight of 500Da to 20000 Da.
3. A process for the preparation of a sulfated derivative of a fucosylated chondroitin sulfate oligosaccharide, comprising the steps of:
(1) Deacetylating fucoidan polysaccharide extracted from body wall and/or viscera of Holothuria nobilis of Echinodermata to obtain intermediate product;
(2) Degrading nitrous acid to obtain fucosylated chondroitin sulfate oligosaccharide dFCS;
(3) The sulfated derivative S-dFCS of the fucosylated chondroitin sulfate oligosaccharide is obtained by subjecting it to a sulfation reaction by a sulfating agent.
4. Use of sulfated derivatives of fucosylated chondroitin sulfate oligosaccharide in preparation of health product or medicine for preventing or treating tumor is provided.
5. The application of sulfated derivative pharmaceutically acceptable salt of fucosylated chondroitin sulfate oligosaccharide in preparing medicaments or health care products for preventing or treating tumors.
6. A medicament for use in anti-tumour, the medicament comprising one or more of a sulfated derivative of fucosylated chondroitin sulfate hexasaccharide, a sulfated derivative of fucosylated chondroitin sulfate nonasaccharide, a sulfated derivative of fucosylated chondroitin sulfate dodecasaccharide, a sulfated derivative of fucosylated chondroitin sulfate pentasaccharide, a sulfated derivative of fucosylated chondroitin sulfate octadeca saccharide.
7. A health product for use in anti-tumour comprising one or more of a sulfated derivative of fucosylated chondroitin sulfate hexasaccharide, a sulfated derivative of fucosylated chondroitin sulfate nonasaccharide, a sulfated derivative of fucosylated chondroitin sulfate dodecasaccharide, a sulfated derivative of fucosylated chondroitin sulfate pentasaccharide and a sulfated derivative of fucosylated chondroitin sulfate octadecanose.
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