Anti-tumor composition and application thereof

Abstract

The invention discloses an anti-tumor composition and application thereof. The active ingredients of the composition are sunitinib with a treatment effective dose and zingiberenone A for enhancing the treatment effect of the sunitinib. The concentration ratio of zingiberenone A to sunitinib is 1: (0.5-2.5). According to the anti-tumor composition provided by the invention, the zingiberenone A and sunitinib are combined for use, so that the drug sensitivity of tumors can be improved, a synergistic effect on inhibiting the multiplication capacity and migration capacity of human kidney cancer cells 786-O is achieved, and a new strategy is provided for treating renal cell carcinoma.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to an antitumor composition and its application. Background Technology

[0002] Renal cell carcinoma (RCC) is the most common malignant tumor of the kidney, accounting for over 90% of all kidney tumors, and is also one of the deadliest types of genitourinary cancer. Currently, targeted therapy with tyrosine kinase inhibitors, represented by sunitinib, has become the first-line treatment for advanced RCC. Sunitinib targets VEGFR2, blocking tumor angiogenesis, thereby improving the survival and prognosis of RCC patients. However, most patients develop resistance after six months of sunitinib use, posing a significant challenge to the clinical management of RCC. Therefore, there is an urgent need to explore new treatment strategies to overcome sunitinib resistance in RCC in order to control its malignant progression.

[0003] Ginger extract, zingerone A, has been explored for the treatment of various diseases, including ulcerative colitis and breast cancer, due to its anti-inflammatory and anti-cancer properties. However, the efficacy of zingerone A in renal cell carcinoma, and whether it can enhance the therapeutic effect of sunitinib in renal cell carcinoma, remains unproven. Summary of the Invention

[0004] The purpose of this invention is to address the shortcomings of the prior art by providing an antitumor composition and its application.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] The first aspect is to provide an antitumor composition, the active ingredient of which is a therapeutically effective dose of sunitinib, and shogaol A for enhancing the therapeutic effect of said sunitinib; the concentration ratio of said shogaol A to sunitinib is 1:(0.5 to 2.5).

[0007] Furthermore, the concentration ratio of sunitinib to gingerone A is 1:2.5.

[0008] More preferably, the concentration of sunitinib is 5–25 μM.

[0009] More preferably, the concentration of gingerone A is 10–50 μM.

[0010] The second aspect is to provide the use of the above composition in the preparation of drugs for treating renal cell carcinoma.

[0011] Furthermore, the drug also includes pharmaceutically acceptable excipients for the composition.

[0012] Furthermore, the excipients include one or more of the following: preservatives, antioxidants, sweeteners, fragrances, adhesives, colorants, surfactants, disintegrants, lubricants, emulsifiers, binders, plasticizers, stabilizers, and suspensions.

[0013] Furthermore, the routes of administration of the drug include one or more of oral, intravenous, intramuscular, and subcutaneous injection.

[0014] The present invention adopts the above technical solution and has the following technical effects compared with the prior art:

[0015] This invention provides an antitumor composition in which gingerone A, when used in combination with sunitinib, can enhance the drug sensitivity of tumors and has a synergistic effect on inhibiting the proliferation and migration of human renal cell carcinoma 786-O cells, providing a new strategy for the treatment of renal cell carcinoma. Attached Figure Description

[0016] Figure 1 This is a graph showing the effect of gingerone A and sunitinib on the drug sensitivity of human renal cell carcinoma cells 786-O in one embodiment of the present invention.

[0017] Figure 2 This is a graph showing the effect of gingerone A and sunitinib on the proliferation of human renal cell carcinoma cells 786-O in one embodiment of the present invention.

[0018] Figure 3 This is a graph showing the effect of gingerone A and sunitinib on the migration ability of human renal cell carcinoma cells 786-O in one embodiment of the present invention. Detailed Implementation

[0019] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but this is not intended to limit the invention. It should be noted that, unless otherwise specified, the embodiments and features described in the embodiments of the present invention can be combined with each other.

[0020] Experimental methods in the following examples, unless otherwise specified, were performed under standard conditions or as recommended by the manufacturer. Unless otherwise stated, all reagents and materials used in the following examples were commercially available.

[0021] Example 1

[0022] This embodiment investigates the effects of gingerone A alone and in combination with sunitinib on the drug sensitivity of human renal cell carcinoma 786-O. The specific experimental steps and structure are as follows:

[0023] 1) Take 786-O cells in the logarithmic growth phase, digest them with trypsin, centrifuge (1000 r / min, 3 min), resuspend and adjust the cell density to 5 × 10⁻⁶. 4Cells / mL. The cell suspension was seeded at 100 μL / well in a 96-well plate and cultured at 37°C and 5% CO2 for 24 h.

[0024] 2) After 24 hours, remove the culture medium and add fresh culture medium containing different concentrations of sunitinib and 12.5 μM gingerone A to the 96-well plate, and continue to treat 786-O cells for 24 hours.

[0025] 3) After 24 hours, add 10 μL of CCK8 reagent to each well and incubate at 37°C for 1 hour.

[0026] 4) Finally, the absorbance at 450 nm was measured using an ELISA reader, the data were analyzed, and the half-maximal inhibitory concentration (IC50) of sunitinib was calculated.

[0027] The results are as follows Figure 1 As shown, gingerone A inhibited the activity of renal cell carcinoma cells, and compared with sunitinib alone, the IC50 of sunitinib decreased and the sensitivity of renal cell carcinoma cells to sunitinib increased after the addition of gingerone A.

[0028] Example 2

[0029] This embodiment investigates the effects of gingerone A alone and in combination with sunitinib on the proliferation of human renal cell carcinoma 786-O. The specific experimental steps and structures are as follows:

[0030] 1) Digest 786-O cells in the logarithmic growth phase with trypsin. After cell detachment, add culture medium to stop the digestion. Collect the cell suspension, centrifuge at 1000 rpm for 3 min, discard the supernatant, and adjust the cell density to 1×10⁶ cells / year with culture medium. 3 Inoculate 100 μL / well into a 96-well plate and incubate at 37°C in a 5% CO2 incubator for 24 h.

[0031] 2) After the culture is completed, remove the old culture medium from the 96-well plate and replace it with a medium that does not contain the drug, contains 5 μM sunitinib, contains 12.5 μM gingerone A, or contains 5 μM sunitinib and 12.5 μM gingerone A, and continue to culture for 24 hours.

[0032] 3) After 24 hours, remove the culture medium, add fresh culture medium containing 10 μM EdU reagent, and incubate for 24 hours.

[0033] 4) After 24 hours, remove the culture medium, fix the cells with 4% paraformaldehyde for 15 minutes, and then wash the cells three times with PBS solution containing 3% BSA.

[0034] 5) Permeate the cells with Triton X-100 for 10 minutes, then wash the cells three times with PBS solution containing 3% BSA.

[0035] 6) Under light-protected conditions, incubate cells with EdU staining reagent at room temperature for 30 minutes. After 30 minutes, wash the cells three times with PBS solution containing 3% BSA.

[0036] 7) Incubate cells with DAPI for 5 minutes, then wash cells 3 times with PBS.

[0037] 8) Use a fluorescence microscope to take pictures and record images of blue light (DAPI) and green light (EdU) respectively, and calculate the cell proliferation rate.

[0038] 9) Calculate the drug synergy index using the formula: MCI = Cell Proliferation Rate A / Cell Proliferation Rate C + Cell Proliferation Rate B / Cell Proliferation Rate C

[0039] Wherein: Cell Proliferation Rate A: Change in cell proliferation rate after treatment with gingerone A;

[0040] Cell Proliferation Rate B: Changes in cell proliferation rate after sunitinib treatment;

[0041] Cell Proliferation Rate C: Changes in cell proliferation rate after simultaneous treatment with gingerone A and sunitinib;

[0042] Based on the above formula, the synergistic index of gingerone A and sunitinib in inhibiting the proliferation of 786-O cells was calculated to be 3.68.

[0043] The results are as follows Figure 2 As shown, sunitinib or shogaol A alone inhibited the proliferation of renal cell carcinoma cells, and the combination of the two drugs showed a stronger inhibitory effect on proliferation, and the inhibitory effect was synergistic.

[0044] Example 3

[0045] This embodiment investigates the effects of gingerone A alone and in combination with sunitinib on the migration ability of human renal cell carcinoma 786-O cells. The specific experimental steps and results are as follows:

[0046] 1) Take 786-O cells in the logarithmic growth phase, digest them with trypsin, centrifuge (1000 rpm, 3 min), resuspend them and seed them in 6-well plates, and culture them at 37℃ and 5% CO2 for 24 h until the cells cover the bottom of the well.

[0047] 2) Take the old culture medium from the 6-well plate, make vertical scratches with a 200 μL sterile pipette tip, wash with PBS to remove detached cells, add drug-free culture medium containing 5 μM sunitinib, 12.5 μM gingerone A, or 5 μM sunitinib and 12.5 μM gingerone A to the 6-well plate, and immediately take pictures under a microscope to record the cell scratches.

[0048] 3) After culturing for another 24 hours, remove the old culture medium from the 6-well plate, wash with PBS to remove dead cells, and photograph the scratch pattern again. Analyze the changes in the scratch pattern between the two photographs and calculate the cell migration rate.

[0049] 4) Calculate the drug synergy index using the formula: MCI = Cell Migration Rate A / Cell Migration Rate C + Cell Migration Rate B / Cell Migration Rate C

[0050] Wherein: Cell Migration Rate A: Change in cell migration rate after treatment with gingerone A;

[0051] Cell Migration Rate B: Changes in cell migration rate after sunitinib treatment;

[0052] Cell Migration Rate C: Changes in cell migration rate after simultaneous treatment with gingerone A and sunitinib;

[0053] Based on the above formula, the synergistic index of gingerone A and sunitinib in inhibiting 786-O cell migration was calculated to be 2.87.

[0054] The results are as follows Figure 3 As shown, sunitinib or shogaol A alone inhibited the migration of renal cell carcinoma cells, while the combined use of the two drugs showed a stronger inhibitory effect on migration and a synergistic effect on the inhibitory effect.

[0055] The above description is merely a preferred embodiment of the present invention and does not limit the implementation and protection scope of the present invention. Those skilled in the art should realize that any equivalent substitutions and obvious changes made based on the content and illustrations of the present invention should be included within the protection scope of the present invention.

Claims

1. An antitumor composition, characterized by comprising, The active ingredient is a therapeutically effective dose of sunitinib and gingerenone A for enhancing the therapeutic effect of the sunitinib; the concentration ratio of the gingerenone A and sunitinib is 1:(0.5-2.5).

2. The composition of claim 1, wherein, The concentration ratio of the sunitinib and gingerenone A is 1:2.

5.

3. Use of the composition of any one of claims 1-2 in the preparation of a medicament for treating renal cell carcinoma.

4. Use according to claim 3, characterized in that, The medicament further comprises a pharmaceutically acceptable excipient of the composition.

5. Use according to claim 4, characterized in that, The excipient comprises one or more of a preservative, antioxidant, sweetener, fragrance, gum base, colorant, surfactant, disintegrant, lubricant, emulsifier, binder, plasticizer, stabilizer, suspending agent.

6. Use according to claim 3, characterized in that, The administration route of the medicament comprises one or more of oral administration, intravenous injection, intramuscular injection, subcutaneous injection.

Images