Application of cordyceps sinensis polysaccharide in preparation of medicine for enhancing tumor treatment effect of PD-1 antibody

Abstract

The invention relates to the technical field of biological medicine, in particular to application of cordyceps sinensis polysaccharide in preparation of a medicine for enhancing the treatment effect of a PD-1 antibody on tumors. According to the invention, the cordyceps sinensis polysaccharide and the PD-1 antibody are combined for use, so that the treatment effect on colorectal cancer is remarkably improved. The combined application of the cordyceps sinensis polysaccharide and the PD-1 antibody not only utilizes the immunoregulation effect of the cordyceps sinensis polysaccharide and the immune checkpoint blocking effect of the PD-1 antibody to improve the tumor inhibition rate, but also discloses the ability of the cordyceps sinensis polysaccharide to regulate the composition and function of intestinal flora, which is a key mechanism for enhancing the curative effect of the PD-1 antibody. According to the invention, a synergistic effect mechanism during combined application is defined for the first time, and a new theoretical basis is laid for future research and drug development.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the use of cordyceps sinensis polysaccharide in the preparation of a drug for enhancing the therapeutic effect of PD-1 antibodies on tumors. Background Art

[0002] Colorectal cancer is a common digestive tract malignancy worldwide, and its occurrence is closely related to genetic, environmental, and lifestyle factors. In the field of tumor immunotherapy, PD-1 antibodies, as immune checkpoint inhibitors, activate T cells and enhance immune responses to tumors by blocking the interaction between PD-1 and PD-L1. However, the efficacy of PD-1 antibodies is limited by individual differences and the tumor microenvironment, and may be accompanied by adverse reactions. Therefore, improving its efficacy is the focus of current research.

[0003] Cordyceps sinensis is a precious traditional Chinese medicine, which is a complex formed by the parasitism of Cordyceps sinensis (Berk.) Sacc. of the family Ergotaceae on the larvae of the insects of the family Bat Moth. Cordyceps sinensis contains a variety of active ingredients, among which Cordyceps sinensis polysaccharide is a natural polysaccharide with multiple biological activities. So far, there has been no report on the combined treatment of colorectal cancer with Cordyceps sinensis polysaccharide and PD-1 antibody. Summary of the invention

[0004] The purpose of the present invention is to provide the use of Cordyceps polysaccharide in the preparation of a drug for enhancing the therapeutic effect of PD-1 antibody on tumors, so as to solve the problems existing in the above-mentioned prior art. The present invention has found that the combined use of Cordyceps polysaccharide and PD-1 antibody can significantly improve the therapeutic effect on tumors.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides the use of Cordyceps sinensis polysaccharide in preparing a medicine for enhancing the therapeutic effect of PD-1 antibody on tumors.

[0007] Furthermore, the tumor is colorectal cancer.

[0008] Furthermore, the preparation method of Cordyceps sinensis polysaccharide comprises the following steps:

[0009] After the Cordyceps sinensis is crushed and decolorized, it is subjected to hot water extraction to obtain a water extract, and then subjected to deproteinization and alcohol precipitation treatment to obtain the Cordyceps sinensis polysaccharide.

[0010] Furthermore, the solid-liquid ratio of the hot water extraction is 1:20.

[0011] The present invention also provides a composition for treating tumors, comprising Cordyceps sinensis polysaccharide and PD-1 antibody.

[0012] Furthermore, the tumor is colorectal cancer.

[0013] Furthermore, the preparation method of Cordyceps sinensis polysaccharide comprises the following steps:

[0014] After the Cordyceps sinensis is crushed and decolorized, it is subjected to hot water extraction to obtain a water extract, and then subjected to deproteinization and alcohol precipitation treatment to obtain the Cordyceps sinensis polysaccharide.

[0015] Furthermore, the solid-liquid ratio of the hot water extraction is 1:20.

[0016] The present invention also provides a medicine for treating tumors, wherein the active ingredient comprises the above-mentioned combination.

[0017] Furthermore, the medicine also includes pharmaceutically acceptable excipients.

[0018] The present invention discloses the following technical effects:

[0019] The present invention significantly improves the therapeutic effect of colorectal cancer by combining Cordyceps polysaccharides with PD-1 antibodies. The combined application of Cordyceps polysaccharides and PD-1 antibodies not only utilizes the immunomodulatory effect of Cordyceps polysaccharides and the immune checkpoint blocking effect of PD-1 antibodies to improve the tumor inhibition rate, but also reveals the ability of Cordyceps polysaccharides to regulate the composition and function of intestinal flora, which is a key mechanism for enhancing the efficacy of PD-1 antibodies. The present invention clarifies for the first time the synergistic mechanism of this combined application, laying a new theoretical foundation for future research and drug development.

[0020] In addition, the composition provided by the present invention is not limited to the treatment of colorectal cancer, but also has the potential to be extended to other tumor types and has broad clinical application prospects.

[0021] The present invention has found that the combined use of Cordyceps polysaccharide and PD-1 antibody can improve the treatment response rate of a specific patient group, especially patients who are insensitive to traditional PD-1 antibody treatment. At the same time, the Cordyceps polysaccharide extraction process provided by the present invention is simple, low-cost, easy to scale production, and conducive to industrial application. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0023] Figure 1 This is the ultraviolet spectrum analysis diagram of Cordyceps sinensis polysaccharide;

[0024] Figure 2 Cordyceps sinensis polysaccharide 1 H spectrum;

[0025] Figure 3 Cordyceps sinensis polysaccharide 13 C spectrum;

[0026] Figure 4 The subcutaneous tumor pictures of mice in different treatment groups in Example 2;

[0027] Figure 5 This is a graph showing the tumor growth curves of mice in different treatment groups in Example 2; * P<0.05, ** P<0.01, *** P < 0.001; ns: P > 0.05;

[0028] Figure 6 This is a statistical analysis of tumor weights of mice in different treatment groups in Example 2; ** P<0.01, *** P < 0.001; ns: P > 0.05;

[0029] Figure 7 CD8 + T cells (A), Treg cells (B), CD8 + T / Treg ratio (C) and IFN-γ + TNF-α + CD8 + Flow cytometry analysis results of T cells (D); * P<0.05, ** P<0.01,

[0030] *** P < 0.001; ns: P > 0.05;

[0031] Figure 8 The results of fluorescence quantitative PCR detection of mice in different treatment groups in Example 2; wherein AD are the expression levels of CD8, Foxp3, granzyme B and perforin, respectively; * P<0.05, ** P<0.01, *** P < 0.001; ns: P > 0.05;

[0032] Fig. 9 The figure is a test result diagram of the intestinal bacterial α diversity of mice in different treatment groups in Example 2; wherein AD represents the ACE index, Chao1 index, Shannon index value and Simpson index respectively; *P<0.05, ** P < 0.01;

[0033] Fig.10 This is the PCoA analysis chart based on the binary_jaccard algorithm;

[0034] Fig.11 This is the PCoA analysis diagram based on the unweighted Unifra algorithm;

[0035] Fig.12 This is the PCA analysis diagram;

[0036] Fig.13 This is the NMDS analysis chart based on the binary_jaccard algorithm;

[0037] Fig.14 This is a gate level analysis diagram of the intestinal flora of mice in different treatment groups in Example 2;

[0038] Fig.15 This is a genus-level analysis diagram of the intestinal flora of mice in different treatment groups in Example 2;

[0039] Fig.16 This is the result of LEfSe analysis and comparison of the difference in intestinal flora of mice in different treatment groups in Example 2;

[0040] Fig.17 The subcutaneous tumor pictures of mice in different treatment groups in Example 3;

[0041] Fig.18 This is a graph showing the tumor growth curves of mice in different treatment groups in Example 3; * P<0.05, ** P<0.01, *** P < 0.001;

[0042] Fig.19 This is a statistical analysis of tumor weights of mice in different treatment groups in Example 3; * P<0.05, *** P<0.001. DETAILED DESCRIPTION

[0043] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0044] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0045] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0046] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0047] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0048] The PD-1 antibody used in the following examples was purchased from Bio X Cell, catalog number: BE0146-50MG.

[0049] Example 1 Extraction of Cordyceps polysaccharide

[0050] 1. Polysaccharide Extraction

[0051] (1) Grinding Cordyceps sinensis to obtain Cordyceps sinensis powder, then adding a decolorizing solution (prepared with methanol, dichloromethane and water in a volume ratio of 4:2:1) and shaking overnight on a shaker, collecting the precipitate by centrifugation, and then drying in an oven at 50° C. until the weight is constant, thereby obtaining decolorized Cordyceps sinensis powder;

[0052] (2) After decolorization, the Cordyceps sinensis powder was soaked in water for 30 minutes and subjected to hot water extraction (a total of 3 extraction cycles, a temperature of 95° C., a solid-liquid ratio of Cordyceps sinensis powder to water of 1:20 (w / v), and a time of 2 hours). The supernatants were combined and concentrated by rotary evaporation to obtain a concentrated solution.

[0053] (3) Deproteinization was performed using Sevag reagent at a volume ratio of 4:1. After oscillation for 30 minutes, the mixture was centrifuged at 4000 r / min for 15 minutes to remove the middle protein layer and the lower organic layer. The upper aqueous phase was subjected to the same operation for 6-7 times until the middle protein layer no longer appeared. The residual organic reagent in the aqueous phase was removed by vacuum rotary evaporation to obtain a crude polysaccharide extract.

[0054] (4) Then, 3 volumes of anhydrous ethanol were added to the crude extract of polysaccharide to precipitate polysaccharide, and after being placed in a refrigerator at 4°C overnight, the extract was centrifuged (3000 r / min, 10 min), the supernatant was removed, and the precipitate was washed with acetone and ether. The precipitate was dissolved in water and dialyzed (to retain polysaccharides with a molecular weight of more than 3500 Da), and then freeze-dried to obtain Cordyceps sinensis polysaccharide.

[0055] 2. Detection and analysis

[0056] The following tests and analyses were performed on the Cordyceps sinensis polysaccharides extracted as described above:

[0057] 2.1 Sugar content analysis

[0058] The polysaccharide content was determined using the phenol-sulfuric acid method. The specific operation was as follows: 0.0, 0.5, 0.75, 1.0, 1.25, 1.5, and 2.0 mL of standard glucose solution (40 μg / mL) were taken into glass test tubes, and the volume was adjusted to 2.0 mL with distilled water. Then 1.0 mL of 5% redistilled phenol solution and 5.0 mL of concentrated sulfuric acid were added, and vortexed to mix. Then the test tubes were placed in a 30°C water bath for 20 minutes. After the reaction was completed, the absorbance was measured at a wavelength of 490 nm. A standard curve was prepared: the x-axis was the glucose concentration and the y-axis was the absorbance.

[0059] Dissolve the sample in distilled water, dilute to a certain concentration, measure the absorbance according to the above method, and calculate the sugar content by comparing with the standard curve. Each sample should be measured at least 3 times.

[0060] 2.2 UV spectrum analysis

[0061] Accurately weigh an appropriate amount of Cordyceps militaris polysaccharide sample, prepare a 1 mg / mL polysaccharide solution with distilled water, use distilled water as a control, scan at 190-500 nm, use wavelength (nm) as the abscissa and the measured absorbance as the ordinate, and draw the UV scanning curve of the polysaccharides of different components of Cordyceps militaris, see Figure 1 .

[0062] 2.3 Protein content detection

[0063] The Bradford protein concentration assay kit produced by Bio-Tech was used to detect the protein content in the polysaccharide.

[0064] 2.4 Monosaccharide composition analysis

[0065] In the experiment, seven monosaccharide standards were first accurately weighed and dissolved, followed by PMP derivatization, including alkali treatment, addition of PMP methanol solution, light-proof water bath, neutralization, dilution, chloroform extraction, and centrifugation, to finally obtain a monosaccharide reference solution. For polysaccharide samples, hydrolysis was first performed, and then the same PMP derivatization steps were used to obtain a sample solution. Both were filtered through a 0.22 μm filter membrane for HPLC analysis.

[0066] 2.5 Nuclear magnetic resonance analysis

[0067] Weigh 20 mg of the sample, dissolve it in 1 mL of D2O, and then freeze-dry it to exchange H with D. The deuteration process was repeated three times. Finally, the polysaccharide was dissolved in 1 mL of D2O and left at room temperature for 3 hours before NMR analysis (see Figure 2-Figure 3 ).

[0068] 3. Experimental results

[0069] The sugar content of Cordyceps polysaccharide was tested by phenol-sulfuric acid method, and the sugar content of Cordyceps polysaccharide was 80.66%. The protein content was 0.18% as detected by Bradford protein concentration assay kit. It can be seen from the full-band UV scanning diagram ( Figure 1 ), Cordyceps polysaccharide showed a strong absorption peak at 200nm, which is the characteristic peak of polysaccharide, while there was no characteristic absorption at 260nm and 280nm, indicating that the extracted Cordyceps polysaccharide contained almost no protein and nucleic acid. The results of monosaccharide composition analysis showed that Cordyceps polysaccharide was mainly composed of galactose, mannose, glucose, xylose and rhamnose, with the composition ratio of mannose (19.4%), rhamnose (2.5%), glucose (5.5%), galactose (59.3%), and xylose (13.3%).

[0070] Example 2: Study on the efficacy of Cordyceps polysaccharide combined with PD-1 antibody in colorectal cancer-bearing mice:

[0071] 1. Experimental Animals

[0072] SPF-grade C57BL / 6 female mice, 6 weeks old and weighing 18±2g, were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd. The experimental animals were raised in an SPF-grade environment in the Experimental Animal Department of Central South University, with free access to water, an ambient temperature of 22±2℃, and an air relative humidity of 60±5%. All animal experiments were conducted in strict accordance with the rules and regulations of the Experimental Animal Ethics Committee of Central South University and international animal welfare and health standards.

[0073] 2. Experimental Methods

[0074] 2.1 Grouping and Dosing

[0075] The mice were randomly divided into six groups: model control group, PD-1 antibody group (PD-1 antibody, 200 μg / mouse), low-dose cordyceps polysaccharide group (CPSL, 50 mg / kg), low-dose cordyceps polysaccharide combined with PD-1 antibody group (PD-1 antibody 200 μg / mouse + CPSL 50 mg / kg), high-dose cordyceps polysaccharide group (CPSH, 150 mg / kg), high-dose cordyceps polysaccharide combined with PD-1 antibody group (PD-1 antibody 200 μg / mouse + CPSH 150 mg / kg). When the inoculated tumor volume reached about 30 mm 3 Cordyceps sinensis polysaccharide was administered orally once a day, and PD-1 antibody treatment was performed simultaneously once every three days, for a total of four times.

[0076] 2.2 Inoculation

[0077] Take colorectal cancer MC38 cells in the logarithmic growth phase and adjust the cell concentration to about 5×10 6 After depilation of the back of C57BL / 6 mice (hereinafter referred to as C57), 100 μL of cell suspension was injected subcutaneously, and then the inoculated C57 mice were kept, the status of the mice was regularly observed and the tumor volume was measured.

[0078] 2.3 Tumor measurements and experimental protocol

[0079] During the administration period, the tumor volume of the mice was measured every three days (the tumor volume calculation formula V = 0.5 × length × width 2 ).

[0080] After the experiment, the mice were killed and the tumor tissue was removed to measure its weight. Part of the tissue was used for subsequent flow cytometry detection, and the remaining tissue could be stored in paraformaldehyde or wrapped in tin foil and frozen in a -80°C refrigerator.

[0081] 2.4 Flow cytometry

[0082] Take the mouse tumor sample, cut it into pieces, grind it through a 70 μm cell sieve, and centrifuge it at 500×g for 10 minutes. Wash the cell pellet twice with PBS, discard the supernatant, and adjust the cell concentration to 10 7 After antibody staining and fixation, resuspend the cells and transfer them to a flow cytometry tube for detection.

[0083] 3. Experimental results

[0084] 3.1 Cordyceps sinensis polysaccharide can enhance the inhibitory effect of PD-1 antibody on MC38 subcutaneous tumor

[0085] like Figure 4-Figure 6As shown in the data, the tumor volume of mice in the model control group increased continuously over time. The use of PD-1 antibodies could significantly reduce the tumor volume and weight of mice and inhibit tumor growth. The combination of Cordyceps polysaccharide and PD-1 antibodies could further reduce the tumor volume compared with the use of PD-1 antibodies alone, and the difference was statistically significant (P<0.05).

[0086] The above results show that Cordyceps polysaccharides can significantly promote the efficacy of PD-1 antibodies.

[0087] 3.2 Cordyceps polysaccharides can enhance the level of immune cell infiltration in mouse tumor tissue

[0088] like Figure 7 As shown, flow cytometry was used to analyze T lymphocytes in the tumor microenvironment of mice. Compared with the PD-1 antibody alone group, the CD8 + T cells increased significantly (P<0.001), while the CD8 + There was no difference in T cells (P>0.05). IFN-γ + TNF-α + CD8 + Compared with the PD-1 antibody monotherapy group, administration of Cordyceps sinensis polysaccharide significantly increased the expression of IFN-γ in tumor tissues. + TNF-α + CD8 + The proportion of T cells (P<0.05). For Treg cells, compared with the Control group, the Treg cells in the CPSH group and the CPSH combined group decreased significantly (P<0.05). Finally, the CD8 + The ratio of T / Tregs ratio, Cordyceps sinensis polysaccharide can increase the CD8 + The ratio of T / Tregs (P<0.001).

[0089] like Figure 8 As shown, real-time fluorescence quantitative PCR experiments were performed to analyze the CD8 + Cordyceps polysaccharides can increase the expression of CD8 in the tumor microenvironment. + T cell infiltration, enhancement of CD8 + It can also inhibit the cytotoxic function of T cells and reduce the number of Tregs in the tumor microenvironment.

[0090] The above results show that Cordyceps sinensis polysaccharides can regulate the immune state of the tumor microenvironment and increase CD8+ T cell infiltration, enhancement of CD8 + T cell toxicity function, enhance CD8 + The ratio of T / Tregs inhibits tumor immune escape.

[0091] 3.3 Effects of Cordyceps polysaccharides on intestinal flora of mice

[0092] like Fig. 9 As shown in the figure, compared with the Control group, the ACE and Chao1 indexes of the PD-1 antibody treatment group were significantly reduced (P<0.05), indicating that PD-1 antibody treatment may lead to a decrease in the total number of intestinal flora species. Compared with the PD-1 antibody group, after CPSL combination, Chao1, ACE and Shannon indexes all showed an upward trend, and Simpson index showed a significant increase (P<0.05), indicating that low-dose Cordyceps polysaccharides can increase the species of intestinal bacteria, the total number of species, and the α diversity. Compared with the PD-1 group, the Chao1, ACE and Shannon indexes of the CPSH combination group also showed an upward trend, indicating that high-dose Cordyceps polysaccharides can also regulate the α diversity of intestinal flora.

[0093] Compared with α diversity, β diversity is mainly used to compare the similarity of species diversity between different samples. Figure 10-13 As shown in the figure, PCoA analysis based on Binary Jaccard algorithm showed that the Control group was not well separated from PD-1, but had a certain separation from CPSH and CPSH combined with PD-1 groups, and the Control group was clearly separated from CPSL and CPSL combined with PD-1 groups. PCoA analysis based on Unweighted UniFrac also confirmed this result. According to the results of principal component analysis (PCA), it can also be seen that the CPSL and CPSL+PD-1 groups are well separated from the Control and PD-1 groups. Then, non-metric multidimensional scaling (NMDS) was used to analyze the changes in the structure of the intestinal flora under different treatments. The results of Binary Jaccard NMDS analysis showed that there was no obvious distinction between the Control group and the PD-1 group, and there was no obvious distinction between the single-dose CPS groups of each dose and the PD-1 group. However, the groups after CPS treatment were well distinguished from the Control group, especially CPSL was completely separated. These results show that CPS can regulate the β diversity of intestinal flora and affect the abundance of flora in the intestine of mice.

[0094] like Fig.14 As shown in Figure 2, at the phylum level, Cordyceps sinensis polysaccharides can change the profile of the intestinal flora of mice, reducing the abundance of Bacteroidetes and increasing the abundance of Firmicutes. Fig.15 As shown in the figure, at the genus level, compared with the PD-1 group, the abundance of Lachnospiraceae_NK4A136 and Faecalibacterium in the CPSL combination group increased, while the abundance of Akkermansia and Lactobacillus decreased. Compared with the PD-1 single-administration group, the abundance of Lactobacillus and Dubosiella in the CPSH combination group increased significantly, while the abundance of Akkermansia, Faecalibacterium and Ileibacterium decreased. In summary, Cordyceps sinensis polysaccharides can regulate the composition and abundance of the intestinal flora of mice. Fig.16 As shown in the figure, the LEfSe analysis results showed that the low-dose CPS enriched Faecalibacterium and two genera of the Lachnospiraceae family (Lachnospiraceae_UCG_006, unidentified_Lachnospiraceae). The high-dose group combined with the high-dose group significantly increased the relative abundance of Lactobacillus and Dubosiella.

[0095] The above results show that Cordyceps sinensis polysaccharides can improve the richness and diversity of intestinal flora in mice, and regulate the composition and blockade of specific bacterial genera.

[0096] Example 3: Study on the efficacy of Cordyceps sinensis polysaccharide combined with PD-1 antibody in a cell line tumor-bearing mouse with PD-1 antibody treatment resistance:

[0097] 1. Experimental Animals

[0098] The animals used in this experiment were 6-week-old male Balb / c mice (SPF grade), weighing 18±2g, purchased from Hunan Slake Jingda Experimental Animal Co., Ltd. and fostered in the Department of Experimental Animals of Central South University. Balb / c mice were raised in an SPF environment and given 12h of light per day. This animal experiment has passed the ethical review of the Department of Experimental Animals of Central South University.

[0099] 2. Experimental Methods

[0100] 2.1 Grouping and Dosing

[0101] Same as “2.1 Grouping and Dosing” in Experimental Example 2.

[0102] 2.2 Inoculation

[0103] Before mice are loaded with tumors, sufficient concentration of 5×10 6CT26.WT cell suspension of 100 μL / mL was added to the mouse, and the hair of the tumor-bearing area was shaved. 100 μL of cell suspension was drawn into the mouse using a 1 mL syringe, and injected subcutaneously into the mouse. The needle was rotated to prevent leakage. After inoculation, the status of the mouse was regularly observed and the tumor volume was measured.

[0104] 2.3 Tumor measurements and experimental protocol

[0105] During the administration period, the tumor volume of the mice was measured every three days (the tumor volume calculation formula V = 0.5 × length × width 2 ).

[0106] After the experiment, the mice were killed and the tumor tissue was removed to measure its weight. Part of the tissue was used for subsequent flow cytometry detection, and the remaining tissue could be stored in paraformaldehyde or wrapped in tin foil and frozen in a -80°C refrigerator.

[0107] 3. Experimental results

[0108] like Figure 17-Figure 19 As shown, compared with the PD-1 antibody group, the tumor volume and weight of mice in the Cordyceps polysaccharide combined with PD-1 antibody group were significantly reduced (P<0.001), and the combination of Cordyceps polysaccharide significantly inhibited tumor growth (P<0.001).

[0109] The above results indicate that Cordyceps sinensis polysaccharides can significantly promote the efficacy of PD-1 antibodies on CT26 subcutaneous tumors.

[0110] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of Cordyceps sinensis polysaccharide in the preparation of drugs for enhancing the therapeutic effect of PD-1 antibodies on tumors.

2. The use according to claim 1, characterized in that: The tumor is colorectal cancer.

3. The use according to claim 1, characterized in that: The preparation method of Cordyceps sinensis polysaccharide comprises the following steps: After the Cordyceps sinensis is crushed and decolorized, it is subjected to hot water extraction to obtain a water extract, and then subjected to deproteinization and alcohol precipitation treatment to obtain the Cordyceps sinensis polysaccharide.

4. The use according to claim 3, characterized in that: The solid-liquid ratio of the hot water extraction is 1:

20.

5. A composition for treating tumors, characterized in that: Including Cordyceps sinensis polysaccharide and PD-1 antibody.

6. The composition according to claim 5, characterized in that The tumor is colorectal cancer.

7. The composition according to claim 5, characterized in that The preparation method of Cordyceps sinensis polysaccharide comprises the following steps: After the Cordyceps sinensis is crushed and decolorized, it is subjected to hot water extraction to obtain a water extract, and then subjected to deproteinization and alcohol precipitation treatment to obtain the Cordyceps sinensis polysaccharide.

8. The composition according to claim 7, characterized in that The solid-liquid ratio of the hot water extraction is 1:

20.

9. A drug for treating tumors, characterized in that: The active ingredient comprises the composition according to claim 5.

10. The drug according to claim 9, characterized in that The drug also includes pharmaceutically acceptable excipients.

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