A composition for treating melanoma

Abstract

The present invention discloses a composition for treating melanoma, belonging to the field of medical technology; the composition for treating melanoma provided by the present invention comprises fangchinoline and dendritic cells. The composition for treating melanoma provided by the present invention has a synergistic effect through fangchinoline and dendritic cells. On the one hand, fangchinoline can enhance the antigen cross-presentation function of dendritic cells, activate the immune system to clear tumors, and achieve autoimmune therapy. On the other hand, dendritic cells can further enhance the anti-tumor effect of fangchinoline itself; thereby obtaining an excellent melanoma treatment effect; and the composition for treating melanoma provided by the present invention has low toxic side effects, can be used to prepare a pharmaceutical preparation, and can also be used in combination with other common drugs or approaches for treating melanoma to improve the treatment effect and prolong the patient's survival.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to a composition for treating melanoma. Background Art

[0002] Melanoma is a highly malignant skin tumor, with an increasing incidence and mortality rate year by year, posing a serious threat to human life and health. Melanoma often occurs on the skin, but can also occur in different parts or tissues such as the mucosa, uvea, and pia mater. Although there are treatment options for melanoma, although melanomas in different parts of the body have certain similarities, the treatment options cannot be used in the same way; for example, conjunctival melanoma and melanoma are both malignant tumors, but they occur in different parts. Conjunctival melanoma is a malignant tumor that occurs on the conjunctiva, while melanoma is generally a malignant tumor that occurs in the skin, mucosa, fundus, etc. Due to the different locations of occurrence, the treatment options cannot be simply transferred.

[0003] Immune surveillance is an important way for the human body to eliminate cancerous cells. When the immune surveillance function is impaired, the malignant transformation of melanocytes cannot be identified and eliminated in time, leading to immune escape and the occurrence of melanoma. Dendritic cells (DCs), as the most powerful antigen-presenting cells, play a key role in human tumor immune surveillance; the loss of DC antigen cross-presentation function is an important reason for tumor cell immune escape. Therefore, it is of great significance to find compounds that can enhance DC antigen cross-presentation function. Summary of the invention

[0004] The purpose of the present invention is to overcome the deficiencies of the above-mentioned prior art and provide a composition for treating melanoma.

[0005] To achieve the above object, in a first aspect of the present invention, the present invention provides a composition for treating melanoma, the composition comprising fangchinoline and dendritic cells.

[0006] A composition for treating melanoma provided by the present invention achieves excellent melanoma treatment effect through the synergistic effect of fangchinoline and dendritic cells (DC cells). In animal experiments, it is found that the obtained composition can significantly reduce the volume and weight of tumors. Specifically, fangchinoline and dendritic cells act synergistically. On the one hand, fangchinoline can enhance the antigen cross-presentation function of dendritic cells, activate the immune system to clear tumors, achieve autologous immune therapy, and avoid immune-related side effects that may be caused by exogenous immunotherapy. On the other hand, dendritic cells can further enhance the anti-melanoma effect of fangchinoline. At the same time, fangchinoline is a natural compound and dendritic cells are also a kind of immune cells. Both have good biocompatibility and low toxic side effects.

[0007] As a preferred embodiment of the composition of the present invention, in the composition, the ratio of dendritic cells to fangchinoline is (1×10 6 -1×10 7 )pcs:(5-15)mg.

[0008] Preferably, in the composition, the ratio of dendritic cells to fangchinoline is (2×10 6 -5×10 6 )pcs:(8-12)mg.

[0009] More preferably, in the composition, the ratio of dendritic cells to fangchinoline is 2.5×10 6 Piece: 10mg.

[0010] The inventors have found that the ratio of dendritic cells and fangchinoline in the provided composition will also affect the performance of the product. When the ratio of the two is within the range given in the present invention, the two can exert a better synergistic effect, and the obtained product has a better therapeutic effect on melanoma.

[0011] As a preferred embodiment of the composition of the present invention, the dendritic cells are monocyte-derived dendritic cells.

[0012] There are differences between dendritic cells from different sources. The inventors have found that when dendritic cells from monocytes are selected, the resulting product has a better effect.

[0013] As a preferred embodiment of the composition of the present invention, the melanoma is skin melanoma.

[0014] Although melanomas in different parts of the body have certain similarities, the treatment methods are different due to the different parts of the body. The inventors have found that the composition provided by the present invention has a better therapeutic effect on skin melanoma.

[0015] In a second aspect of the present invention, the present invention provides a pharmaceutical preparation for treating melanoma, the pharmaceutical preparation comprising fangchinoline and dendritic cells.

[0016] As a preferred embodiment of the pharmaceutical preparation of the present invention, the dendritic cells are monocyte-derived dendritic cells.

[0017] As a preferred embodiment of the pharmaceutical preparation of the present invention, the melanoma is skin melanoma.

[0018] As a preferred embodiment of the pharmaceutical preparation of the present invention, in the pharmaceutical preparation, the dosage of fangchinoline is (5-15) mg / kg, and the dosage of dendritic cells is (1×10 6-1×10 7 ) pieces / kg.

[0019] Preferably, in the pharmaceutical preparation, the dosage of fangchinoline is (8-12) mg / kg, and the dosage of dendritic cells is (2×10 6 -5×10 6 ) pieces / kg.

[0020] More preferably, in the pharmaceutical preparation, the dosage of fangchinoline is 10 mg / kg, and the dosage of dendritic cells is 2.5×10 6 Pieces / kg.

[0021] As a preferred embodiment of the pharmaceutical preparation of the present invention, the pharmaceutical preparation further comprises a pharmaceutically acceptable carrier.

[0022] As a preferred embodiment of the pharmaceutical preparation of the present invention, the dosage form of the pharmaceutical preparation is an injection.

[0023] In the third aspect of the present invention, the present invention provides a pharmaceutical composition for treating melanoma, wherein the pharmaceutical composition comprises the composition for treating melanoma of the present invention, and other commonly used drugs for treating melanoma.

[0024] As a preferred embodiment of the pharmaceutical composition of the present invention, the other commonly used drugs for treating melanoma include at least one of paclitaxel, cisplatin, temozolomide, dacarbazine, carboplatin and vemurafenib.

[0025] As a preferred embodiment of the pharmaceutical composition of the present invention, the dendritic cells are monocyte-derived dendritic cells.

[0026] As a preferred embodiment of the pharmaceutical composition of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0027] In the fourth aspect of the present invention, the present invention provides the use of fangchinoline and dendritic cells in combination for preparing a drug for treating melanoma.

[0028] As a preferred embodiment of the application of the present invention, the dendritic cells are monocyte-derived dendritic cells.

[0029] As a preferred embodiment of the use of the present invention, the melanoma is skin melanoma.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] A composition for treating melanoma provided by the present invention has a synergistic effect through fangchinoline and dendritic cells. On the one hand, fangchinoline can enhance the antigen cross-presentation function of dendritic cells, activate the immune system to clear tumors, and achieve autoimmune therapy. On the other hand, dendritic cells can further enhance the anti-tumor effect of fangchinoline itself, thereby obtaining an excellent melanoma treatment effect. In addition, the composition for treating melanoma provided by the present invention has low toxic and side effects, can be used to prepare a pharmaceutical preparation, and can also be used in combination with other common drugs or approaches for treating melanoma to improve the treatment effect and prolong the patient's survival. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is a graph showing the effect of fangchinoline on antigen cross-presentation function in Example 1;

[0033] Figure 2 This is a comparison chart of the tumor mass results in Example 2. DETAILED DESCRIPTION

[0034] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0035] Unless otherwise specified, the reagents and equipment used in the present invention are all conventionally available.

[0036] Example 1

[0037] The present invention explores the effect of fangchinoline on antigen cross-presentation function, which specifically includes the following steps:

[0038] Experimental steps: DC cell line MutuDC was used to flow cytometry to detect the effect of fangchinoline (Fan) on MutuDC antigen cross-presentation. Grouping: negative, IgG, Ctrl, Fan 5μM, 1 million MutuDC were plated per well in a six-well plate, Fan and OVA protein were added the next day, OVA protein 100ug / mL was added to all four wells, and Fan 5μM was added to the last group. After 24 hours of action, the OVA257-264 (SIINFEKL) peptide is an MHC class I molecule (H-2K in mice). b )-restricted OVA peptide antigen epitope, MutuDC cell surface H-2K b The amount of / SIINFEKL complexes represents the ability of MutuDC to cross-present antigens. b The fluorescent antibody staining of the / SIINFEKL complex was followed by flow cytometry detection. The stronger the fluorescence intensity, the stronger the antigen cross-presentation ability of MutuDC.

[0039] The results are as follows Figure 1 As shown, dendritic cells treated with fangchinoline can significantly enhance their presentation of OVA protein.

[0040] Example 2

[0041] The present invention explores the therapeutic effect of fangchinoline and dendritic cells on mouse melanoma, which specifically includes the following steps:

[0042] (1) A mouse xenograft tumor model was established by inoculating C57BL / 6 mice with B16 cells (mouse melanoma cells);

[0043] (2) The mice with the successfully constructed melanoma transplanted tumor model were grouped into Ctrl, DC, Fan, and DC+Fan groups, with 8 mice in each group. Ctrl was a blank group, which was neither inoculated with DC cells nor treated with Fan; Example 1 was both inoculated with DC cells and treated with Fan; Comparative Example 1 was only inoculated with DC cells without Fan treatment; Comparative Example 2 was only treated with Fan without DC cell inoculation. Three days before inoculation of DC cells, the groups corresponding to Example 1 and Comparative Example 2 were intraperitoneally injected with corresponding doses of fangchinoline, and the injection was continued for three consecutive days, once a day. Comparative Example 1 and the blank group were injected with normal saline at a dose of 10 mg / kg at the same time; after three consecutive injections, the groups corresponding to Example 1 and Comparative Example 1 were inoculated with DC cells according to the dose; Example 1 and Comparative Example 2 continued to inject fangchinoline daily according to the dose in the table until the sample was obtained. On the 6th day, the tumor volume was recorded every day. On the 15th day, the tumor volume of each group was measured first, and then the tumor was taken out and photographed.

[0044] Table 1

[0045] Fangchinoline Monocyte-derived DCs Blank Group 0 0 Example 1 10mg / kg <![CDATA[2.5×10 6 Piece / kg<!-- 3 --> ]]> Comparative Example 1 0 <![CDATA[2.5×10 6 pcs / kg]]> Comparative Example 2 10mg / kg 0

[0046] (3) The tumor volume was measured and recorded every day starting from the 6th day after inoculation. The tumor was sampled and dissected 15 days after inoculation, and the tumor weight was measured and recorded in Table 2, where nd represents the nth day after inoculation of melanoma cells, and the tumor weight is the weight of the sample taken 15 days after inoculation of melanoma cells, after the tumor was dissected, cleaned, and dried; all values ​​are the average values ​​of each group.

[0047] Table 2

[0048] <![CDATA[0d(mm 3 )]]> <![CDATA[6d(mm 3 )]]> <![CDATA[10d(mm 3 )]]> <![CDATA[15d(mm 3 )]]> Tumor weight (g) Blank Group 0 30.75 196 927 1.2539 Example 1 0 9.26 66 269 0.7348 Comparative Example 1 0 25 193 967 1.4089 Comparative Example 2 0 34 104 951 1.4076

[0049] As can be seen from Table 2, when the technical solution of the present invention (Example 1) is adopted, the growth rate of the tumor can be significantly controlled, and the tumor volume increases slowly after inoculation. The average volume of the tumor is 269 mm at 15 days after inoculation. 3The average tumor weight was 0.735 g, and compared with the blank group, the volume reduction was more than 70%, and the tumor weight reduction was more than 40%; it can be seen from Example 1 and Comparative Examples 1-2 that when fangchinoline or dendritic cells are not added, the anti-tumor effect cannot be achieved; wherein Figure 2 The pictures of melanoma tumor masses in the blank group (Ctrl), Example 1 (Fan+DC), Comparative Example 1 (DC) and Comparative Example 2 (Fan) were taken 15 days later. It can be seen intuitively from the picture that only under the technical solution provided by the present invention can the volume of the tumor be effectively reduced and an excellent therapeutic effect be achieved.

[0050] Finally, it should be noted that the above embodiments are intended to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. A composition for treating melanoma, characterized in that: The composition comprises fangchinoline and dendritic cells; In the composition, the ratio of dendritic cells to fangchinoline is (1×10 6 -1×10 7 )pcs:(5-15)mg; The dendritic cells are monocyte-derived dendritic cells; The melanoma is skin melanoma.

2. A pharmaceutical preparation for treating melanoma, characterized in that: The pharmaceutical preparation comprises fangchinoline and dendritic cells.

3. The pharmaceutical preparation according to claim 2, characterized in that In the pharmaceutical preparation, the dosage of fangchinoline is (5-15) mg / kg, and the dosage of dendritic cells is (1×10 6 -1×10 7 ) pieces / kg.

4. The pharmaceutical preparation according to claim 2, characterized in that The pharmaceutical preparation also includes a pharmaceutically acceptable carrier.

5. The pharmaceutical preparation according to claim 2, characterized in that The dosage form of the pharmaceutical preparation is injection.

6. A pharmaceutical composition for treating melanoma, characterized in that: The pharmaceutical composition comprises the composition for treating melanoma as claimed in claim 1, and other commonly used drugs for treating melanoma.

7. Application of fangchinoline and dendritic cells in combination for the preparation of drugs for the treatment of melanoma.

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