Reagent for inhibiting pancreatic cancer cell proliferation and application thereof

Abstract

The invention belongs to the technical field of pancreatic cancer cell inhibitors, and particularly relates to a reagent for inhibiting pancreatic cancer cell proliferation and application thereof. The reagent for inhibiting pancreatic cancer cell proliferation is prepared by mixing rapamycin, isopropyl myristate and levulinic acid according to the mass ratio of 10: (1-2): (0.1-1). The invention develops a reagent for inhibiting the proliferation of pancreatic cancer cells, and the effect of inhibiting the proliferation of MIA PaCa-2 and PANC-1 pancreatic cancer cells is achieved by inhibiting the expression of NKAP.

Description

Technical Field

[0001] This invention belongs to the field of pancreatic cancer cell inhibitor technology, specifically relating to a reagent for inhibiting the proliferation of pancreatic cancer cells and its application. Background Technology

[0002] Pancreatic cancer suffers from low survival rates due to the long-term interaction of multiple factors, including genetic factors, environmental factors, and molecular biological abnormalities. Molecular biological studies have shown that pancreatic cancer is associated with abnormalities in genes such as BRCA1 / BRCA2, CDKN2A, STK11, PALB2, and KRAS. For example, the mutation rate of the KRAS gene in pancreatic cancer is over 90%. Mutated KRAS genes continuously activate downstream signaling pathways, leading to uncontrolled cell proliferation and differentiation.

[0003] A crucial factor in pancreatic cancer treatment is inhibiting the proliferation of pancreatic cancer cells. Current research on agents that inhibit pancreatic cancer cell proliferation focuses on various gene therapy targets. For example, sotorasiib inhibits pancreatic cancer cells through the KRAS G12C-related pancreatic cancer treatment target; erlotinib combined with gemcitabine can inhibit pancreatic cancer cells by suppressing GFR / MEK / STAT; and PD-1 inhibitors can also inhibit pancreatic cancer cell proliferation to some extent.

[0004] Although the aforementioned existing technologies have investigated the effects of different targets on the treatment of pancreatic cancer and have studied pancreatic cancer cell inhibitory products targeting different therapeutic targets, the emergence of drug resistance can affect the inhibitory effect on pancreatic cancer cells, thereby impacting the overall treatment efficacy. Therefore, there is still a need to develop new pancreatic cancer cell inhibitory products. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides a reagent for inhibiting the proliferation of pancreatic cancer cells and its application.

[0006] The purpose of this invention is to provide a reagent for inhibiting the proliferation of pancreatic cancer cells, which is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1~2:0.1~1.

[0007] Rapamycin is a macrolide compound that was first isolated from soil bacteria and has immunosuppressive and antitumor effects.

[0008] Isopropyl myristate is a fatty acid ester and is stable.

[0009] Acetopropionic acid is an organic acid containing ketone and carboxyl groups.

[0010] When rapamycin, isopropyl myristate, and levulinic acid are thoroughly mixed, they can remain relatively stable under normal temperature, pressure, and light-protected conditions, which facilitates the exertion of their medicinal effects.

[0011] Preferably, the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells refers to pancreatic cancer cells that are MIA PaCa-2 and / or PANC-1.

[0012] Preferably, the above-mentioned reagents for inhibiting the proliferation of pancreatic cancer cells are used to prepare an anti-pancreatic cancer drug, which is compounded in equal mass proportions.

[0013] Preferably, in the application of the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells, the drug has the reagent for inhibiting the proliferation of pancreatic cancer cells as its sole active ingredient.

[0014] Preferably, in the application of the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells, the active ingredient of the drug is a reagent for inhibiting the proliferation of pancreatic cancer cells and a known anti-pancreatic cancer drug.

[0015] Preferably, in the application of the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells, the known anti-pancreatic cancer drug is at least one of gemcitabine, albumin-bound paclitaxel, capecitabine, irinotecan, oxaliplatin, erlotinib, and olaparib.

[0016] Preferably, in the application of the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells, the drug contains pharmaceutically acceptable excipients.

[0017] Preferably, in the application of the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells, the pharmaceutically acceptable excipients include at least one of lipid-soluble solvents, co-solvents, oily carriers, solubilizers, emulsifiers, dispersants, binders, diluents, adhesives, disintegrants, lubricants, and preservatives.

[0018] Preferably, in the application of the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells, the dosage form of the drug is a capsule, and the appropriate pharmaceutically acceptable excipients are diluents (such as lactose, microcrystalline cellulose, mannitol), binders (such as povidone, hydroxypropyl methylcellulose, gelatin), disintegrants (such as sodium crosyl carboxymethyl cellulose, crosyl povidone), lubricants (such as magnesium stearate, talc, polyethylene glycol 6000), and preservatives (such as benzyl alcohol, parabens).

[0019] Preferably, in the application of the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells, the dosage form of the drug is a solid dosage form, and the appropriate pharmaceutically acceptable excipients are dispersants and binders. Examples include polyvinylpyrrolidone, povidone, hydroxypropyl methylcellulose, β-cyclodextrin, and starch.

[0020] Preferably, in the application of the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells, the dosage form of the drug is a liquid, and the appropriate excipients are emulsifiers, solubilizers, dispersants, lipid-soluble solvents, and co-solvents. For example, phospholipids (hydrogenated soybean phosphatidylcholine, distearyl phosphatidylcholine), cholesterol, distearyl phosphatidylethanolamine-polyethylene glycol 2000, Tween-80, and poloxamer 188 can be used as excipients to prepare liposomes, microemulsions, or nanoemulsions in the form of a liquid emulsion.

[0021] Preferably, in the application of the above-mentioned reagent for inhibiting the proliferation of pancreatic cancer cells, the active ingredient in the drug has a mass percentage of 10 mg / g to 50 mg / g.

[0022] Compared with the prior art, the present invention has the following beneficial effects: The core innovation of this invention lies in the development of a reagent that inhibits the proliferation of pancreatic cancer cells, composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1~2:0.1~1. This reagent inhibits the proliferation of MIA PaCa-2 and PANC-1 pancreatic cancer cells by suppressing NKAP expression. This invention clarifies for the first time the relationship between rapamycin, isopropyl myristate, and levulinic acid and NKAP, and also clarifies for the first time the relationship between rapamycin, isopropyl myristate, and levulinic acid and pancreatic cancer.

[0023] Based on the same inventive concept, this invention provides examples of formulations for preparing antitumor drugs using the reagent that inhibits the proliferation of pancreatic cancer cells, such as solid dosage forms, liquid dosage forms, or capsule dosage forms, and also provides exemplary types of excipients. These practically applicable drug preparation techniques rely on the reagent that inhibits the proliferation of pancreatic cancer cells; that is, the active ingredients all contain the reagent that inhibits the proliferation of pancreatic cancer cells. Therefore, these drugs also have anti-pancreatic cancer cell effects.

[0024] Furthermore, for the purpose of treating diseases, the reagent of the present invention for inhibiting the proliferation of pancreatic cancer cells can also be combined with other known pancreatic cancer treatment drugs to enhance the therapeutic effect. These known anti-pancreatic cancer drugs are at least one of gemcitabine, albumin-bound paclitaxel, capecitabine, irinotecan, oxaliplatin, erlotinib, and olaparib. Attached Figure Description

[0025] Figure 1 The OD values ​​of tumor cell sap after knockdown or overexpression of NKAP are shown. A represents MIA PaCa-2 and its knockdown cells, B represents cells related to MIA PaCa-2 overexpression, C represents PANC-1 and its knockdown cells, and D represents cells related to PANC-1 overexpression.

[0026] Figure 2This is a colony formation assay for pancreatic cancer cells after NKAP knockdown. Row A shows the results for MIA PaCa-2 cells, and row B shows the results for PANC-1 cells.

[0027] Figure 3 This is an assay for colony formation of pancreatic cancer cells after overexpression of NKAP. Row A shows the results for MIA PaCa-2 cells, and row B shows the results for PANC-1 cells.

[0028] Figure 4 For MIA PaCa-2 drug experiments.

[0029] Figure 5 For PANC-1 drug experiments.

[0030] Figure 6 This is a graph showing the results of a Western blotting analysis. A represents MIA PaCa-2 cells, and B represents PANC-1 cells. Detailed Implementation

[0031] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings.

[0032] Unless otherwise specified, all reagents used in this invention are commercially available, and all methods used are conventional techniques in the art.

[0033] The inventive concept of this invention is as follows: NKAP (NF-κB Activating Protein) is a multifunctional protein associated with various diseases. For example, in studies of lung cancer related to smoke exposure, NKAP expression levels are correlated with the phosphorylation status of the MARCKS protein; missense mutations in the NKAP gene can lead to phenotypic abnormalities and induce hereditary diseases; furthermore, research on NKAP in gliomas is still in its early stages, but it is known to participate in tumorigenesis by regulating the NF-κB signaling pathway. However, the relationship between NKAP and pancreatic cancer is not yet clear. Furthermore, there is an urgent need in this field to develop new therapeutic drugs for pancreatic cancer.

[0034] Therefore, the present invention provides a reagent for inhibiting the proliferation of pancreatic cancer cells, which is composed of rapamycin, isopropyl myristate and levulinic acid in a mass ratio of 10:1~2:0.1~1.

[0035] Rapamycin is a macrolide compound (CAS number 53123-88-9), first isolated from soil bacteria, and possesses immunosuppressive and antitumor effects. Existing research indicates that it regulates cell growth, proliferation, and metabolism by inhibiting the mTOR signaling pathway, making it a focus of research in organ transplantation, cancer treatment, and anti-aging. The results of this invention show that rapamycin can inhibit NKAP expression to a certain extent, clarifying the association between rapamycin and pancreatic cancer.

[0036] Isopropyl myristate is a stable fatty acid ester (CAS number 110-27-0). It is easily absorbed by cells and readily mixes with other substances. In the reagent for inhibiting pancreatic cancer cell proliferation in this invention, it acts as a dispersant and absorption promoter, while also slightly inhibiting NKAP expression, thus contributing to the inhibition of pancreatic cancer cell proliferation.

[0037] Acetopropionic acid, an aliphatic keto acid (CAS number 123-76-2), is commonly used as an intermediate in drug synthesis, such as in the production of anti-inflammatory and analgesic drugs. Its calcium salt can also promote bone formation. In the reagent of this invention for inhibiting pancreatic cancer cell proliferation, prolonged (72h) action can slightly inhibit NKAP expression, laying the foundation for inhibiting pancreatic cancer cell proliferation.

[0038] This invention clarifies for the first time the relationship between rapamycin, isopropyl myristate, and levulinic acid and NKAP, and also clarifies for the first time the relationship between rapamycin, isopropyl myristate, and levulinic acid and pancreatic cancer. Furthermore, the reagent of this invention for inhibiting pancreatic cancer cell proliferation, with rapamycin, isopropyl myristate, and levulinic acid as active ingredients, when combined, can significantly inhibit NKAP expression and inhibit the proliferation of pancreatic cancer cells.

[0039] Example 1 An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1:0.1.

[0040] Example 2 An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:2:1.

[0041] In this invention, the reagent that inhibits the proliferation of pancreatic cancer cells achieves its effect by suppressing the expression of NKAP. The mechanism and the verification of the inhibitory effect are presented below.

[0042] Experiment 1: Construction of NKAP knockdown and overexpression cell models Using molecular biology techniques such as gene synthesis, siRNA interference, and vector construction, NKAP was knocked down in MIA PaCa-2 and PANC-1 pancreatic cancer cells, resulting in low-expression cell models. Conversely, NKAP was overexpressed in both MIA PaCa-2 and PANC-1 pancreatic cancer cells, resulting in overexpression cell models.

[0043] Specifically, the methods for constructing the MIA PaCa-2 knockdown negative control cell line (si-NC cells), knockdown cell line (si NKAP-1), overexpression cell line (over-NKAP), and overexpression negative control cell line (over-NC) are as follows: Based on the NKAP nucleic acid sequence (NM_024528.4), specific siRNA was designed and synthesized to construct a knockdown model, and the overexpression model used the commercial pEX-3-NKAP plasmid. si-NC and pEX-1 empty vectors were used as knockdown negative controls and overexpression negative controls, respectively. Transient cell transfection was performed, and mRNA levels were detected by qRT-PCR after 24 h, and protein levels were detected by Western Blot after 48 h. Knockdown negative control cell lines, knockdown cell lines, overexpression cell lines, and overexpression negative control cell lines were screened.

[0044] The construction methods for the PANC-1 knockdown negative control cell line (si-NC cells), knockdown cell line (si NKAP-1), overexpression cell line (over-NKAP), and overexpression negative control cell line (over-NC) were the same as those for MIA PaCa-2.

[0045] The si-NC sequence is as follows: 5'-UUCUCEGAACGUGUCACGUTTACGUGACACGUUCGGAGAATT-3'.

[0046] The si NKAP-1 sequence is as follows: 5'-CUGCGGCAGAAGAGAUUAATTUUAAUCUCUUCUGCCGCAGTT-3'.

[0047] Experiment 2: Cell proliferation-related experiments (1) CCK-8 cell proliferation experiment (1.1) Collect MIA PaCa-2 pancreatic cancer cells in good growth condition, digest them, and resuspend them in a culture medium without antibiotics. The cell concentration was measured by a cell counting chamber and adjusted to 1000 cells / 100μL of MIA PaCa-2 cell suspension.

[0048] (1.2) After thoroughly mixing the above MIA PaCa-2 cell suspension, seed it into a 96-well plate with 5 replicates per group. Leave blank wells with the same number of sample wells when laying the plate.

[0049] (1.3) After culturing for 24 hours, the cells were transfected, and the culture medium in the 96-well plate was replaced 6 hours after transfection. The materials used for transfection were the siRNA described in Experiment 1, the commercial pEX-3-NKAP plasmid, si-NC, and pEX-1 empty vector. This step of the culture was carried out in the dark.

[0050] (1.4) At 0h, 24h, 48h and 72h after transfection, 10μL of CCK-8 reagent was added to each well. After reacting in the dark for 2h, the absorbance of each well was measured at 450nm using a microplate reader. In addition, a control group (Mock) of MIA PaCa-2 cells was prepared.

[0051] The CCK-8 cell proliferation assay for PANC-1 was performed as described in (1.1) to (1.4) above.

[0052] OD values ​​of the fluid from two types of pancreatic cancer cells are shown in the attached table. Figure 1 .

[0053] Figure 1 In MIA PaCa-2, knocking down NKAP significantly reduced the OD value of the cell sap, indicating that it could inhibit the proliferation of the cells. Figure 1 In cell line B, overexpression of NKAP in MIA PaCa-2 significantly increased the OD value of the cell sap, indicating that it can promote the proliferation of this cell. A similar pattern was observed with MIA PaCa-2. Figure 1 In the C group, knocking down NKAP in PANC-1 significantly reduced the OD value of the cell sap, indicating that it could inhibit the proliferation of the cell. Figure 1 In the D-cell, overexpression of NKAP in PANC-1 significantly increased the OD value of the cell sap, indicating that it can promote the proliferation of the cell. Figure 1 *** indicates a significant difference compared to the control group (i.e., the Mock). p <0.001, Mock represents primitive pancreatic cancer cells, si-NC represents the negative control with knockdown, si NKAP-1 represents NKAP knockdown, over-NC represents the empty plasmid control with overexpression, and over-NKAP represents NKAP overexpression.

[0054] Figure 1 The results suggest that NKAP expression is significantly correlated with the proliferation of pancreatic cancer cells and can serve as a therapeutic target. When the goal is to treat pancreatic cancer and inhibit its proliferation, NKAP expression should be suppressed.

[0055] (2) Plate colony formation experiment (2.1) Cell seeding: After digesting the MIA PaCa-2 pancreatic cancer cells in the culture dish, the cells were seeded into a six-well plate at a density of 1 × 10⁶ cells per well. 6 Cells were cultured at 37°C and 5% CO2 for 24 hours before transfection. The medium was changed in the six-well plates after 6 hours. The materials used for transfection were the siRNA described in Experiment 1, the commercial pEX-3-NKAP plasmid, si-NC, and pEX-1 empty vector. This step of the culture was performed in the dark.

[0056] (2.2) 24 h after transfection, the cells in the six-well plate were digested and seeded into 12-well plates, 1000 cells per well. The cells were then placed in a 37°C, 5% CO2 incubator and cultured in the dark for 9 days to form clones. The culture medium was changed every 3 days.

[0057] (2.3) After the clones are formed and visible to the naked eye, the culture medium is aspirated, the cells are washed twice with PBS, fixed with 0.01% paraformaldehyde for 30 min, and then washed once with PBS.

[0058] (2.4) After staining with 4% crystal violet for 15 min and washing once with PBS, the 12-well plate was inverted and air-dried, and photographed and recorded. The results were quantitatively analyzed using ImageJ software. In addition, a control group (Mock) of MIA PaCa-2 cells was prepared.

[0059] The plate colony formation experiment of PANC-1 was carried out in accordance with (2.1) to (2.4) above.

[0060] See results Figure 2 and Figure 3 , Figure 2 The results showed that after knocking down NAKP, the number of MIA PaCa-2 and PANC-1 pancreatic cancer cells was significantly reduced compared with the original pancreatic cancer cell group. Figure 2 *** indicates a significant difference compared to the control group (i.e., the original tumor cells). p <0.001, Mock represents primitive pancreatic cancer cells, si-NC represents the knockdown negative control, and si NKAP-1 represents the knockdown of NKAP.

[0061] Figure 3 The results showed that after NAKP overexpression, the number of MIA PaCa-2 and PANC-1 pancreatic cancer cells was significantly increased compared with the original pancreatic cancer cell group. Figure 3 *** indicates a significant difference compared to the control group (i.e., the original tumor cells). p <0.001, Mock represents primitive pancreatic cancer cells, over-NC represents overexpression of empty plasmid control, and over-NKAP represents overexpression of NKAP.

[0062] Figure 2 and Figure 3 The results also suggest that NKAP expression is significantly correlated with the proliferation of pancreatic cancer cells and can serve as a therapeutic target. When the goal is to treat pancreatic cancer and inhibit the proliferation of pancreatic cancer cells, NKAP expression should be suppressed.

[0063] Experiment 3, Drug Experiment (1) Grouping of MIA PaCa-2 cells Blank control group 1: MIA PaCa-2+ dimethyl sulfoxide.

[0064] Blank control group 2: PANC-1 + dimethyl sulfoxide.

[0065] Experimental Group 1: The reagent from Example 1 was dissolved in dimethyl sulfoxide, and the reagent concentration was 10 mg / mL.

[0066] Experimental Group 2: The reagent from Example 2 was dissolved in dimethyl sulfoxide, and the reagent concentration was 10 mg / mL.

[0067] Single-factor control 1: Veuvulinic acid was dissolved in dimethyl sulfoxide, and the concentration of levulinic acid was prepared as 1 mg / mL.

[0068] Single-factor control 2: Isopropyl myristate was dissolved in dimethyl sulfoxide, and the concentration of isopropyl myristate was prepared as 0.1 mg / mL.

[0069] Single-factor control 3: Rapamycin was dissolved in dimethyl sulfoxide, and the concentration of rapamycin was 10 mg / mL.

[0070] (2) Grouping of PANC-1 cells Blank control group 2: PANC-1 + dimethyl sulfoxide.

[0071] Experimental Group 3: The reagent from Example 1 was dissolved in dimethyl sulfoxide, and the reagent concentration was 10 mg / mL.

[0072] Experimental Group 4: The reagent from Example 2 was dissolved in dimethyl sulfoxide, and the reagent concentration was 10 mg / mL.

[0073] Single-factor control 4: levulinic acid was dissolved in dimethyl sulfoxide to prepare a concentration of 0.1 mg / mL.

[0074] Single-factor control 5: Isopropyl myristate was dissolved in dimethyl sulfoxide to prepare a concentration of 1 mg / mL.

[0075] Single-factor control 6: Rapamycin was dissolved in dimethyl sulfoxide to prepare a concentration of 10 mg / mL.

[0076] The CCK-8 cell proliferation experiment was conducted according to the method in Experiment 2: Tumor cells in good growth condition were collected, digested, and resuspended in culture medium without antibiotics for MIA PaCa-2 and PANC-1 pancreatic cancer cells. The cell concentration was determined by cell counting chamber and adjusted to 1000 cells / 100 μL of cell suspension. The cell suspension was thoroughly mixed and seeded into 96-well plates with 5 replicates per group. Blank wells were reserved with the same number of sample wells during plate formation. After 24 h of culture, the drug was added to the cells. The specific grouping settings were as described in Experiment 3 (1) above. The culture medium in the 96-well plate was replaced after 6 h. This step of culture was carried out in the dark. 4) At 0 h, 24 h, 48 h, and 72 h after adding the drug, 10 μL of LCK-8 reagent was added to each well. After reacting in the dark for 2 h, the absorbance of each well was detected at a wavelength of 450 nm using an ELISA reader.

[0077] See results Figure 4 and Figure 5 The results showed that the agents in Examples 1 and 2 exhibited the strongest inhibitory effect on pancreatic cancer cells, regardless of whether they were targeting MIA PaCa-2 or PANC-1 pancreatic cancer cells. While single-factor controls 1 and 2 showed varying degrees of inhibitory effect, the effect was slightly weaker. Although there were significant differences between single-factor control group 3 and blank control group 1, and between single-factor control group 6 and blank control group 2, the effects of Examples 1 and 2 were more pronounced.

[0078] Figure 4 and Figure 5 In the figure, ** indicates a significant difference (P < 0.01), and *** indicates a significant difference (P < 0.001). It should be noted that differences greater than *** still indicate significant differences (e.g., at 48h, there were significant differences between the blank control group 1, the single-factor control group 1, and the single-factor control group 2 and the experimental group 1, P < 0.001). Due to the large number of lines in the figure, they are not labeled.

[0079] Western blotting was used to detect NKAP expression levels, with GAPDH as an internal control. Results are shown below. Figure 6 .

[0080] The results showed that rapamycin, isopropyl myristate, and levulinic acid could all inhibit NKAP expression.

[0081] Combination Figures 4-6 The results showed that rapamycin, isopropyl myristate, and levulinic acid could all inhibit the proliferation of MIA PaCa-2 and PANC-1 pancreatic cancer cells by suppressing NKAP expression.

[0082] It should be noted that the above-described Examples 1 and 2 are merely illustrative examples of drug demonstration and experimental results, and should not be construed as limiting the present invention. When numerical ranges are involved in the present invention, it should be understood that both endpoints of each numerical range, as well as any value between the two endpoints, can be selected, since the steps and methods used are the same as in the examples. Although preferred embodiments of the present invention have been described, those skilled in the art, once they understand the inventive concept of the present invention, can make other changes and modifications to these embodiments, and these changes and modifications all fall within the scope of the present invention. For example, the technical solution of the present invention can also be extended to the following embodiments, all of which have the effect of inhibiting the proliferation of MIA PaCa-2 and PANC-1 pancreatic cancer cells by inhibiting the expression of NKAP.

[0083] Example 3 This embodiment involves adjusting the dosage of isopropyl myristate.

[0084] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1.2:1.

[0085] Example 4 This embodiment involves adjusting the dosage of isopropyl myristate.

[0086] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1.4:1.

[0087] Example 5 This embodiment involves adjusting the dosage of isopropyl myristate.

[0088] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1.6:1.

[0089] Example 6 This embodiment involves adjusting the dosage of isopropyl myristate.

[0090] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1.8:1.

[0091] Example 7 This example describes the adjustment of the dosage of levulinic acid.

[0092] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1:0.2.

[0093] Example 8 This example describes the adjustment of the dosage of levulinic acid.

[0094] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1:0.4.

[0095] Example 9 This example describes the adjustment of the dosage of levulinic acid.

[0096] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1:0.6.

[0097] Example 10 This example describes the adjustment of the dosage of levulinic acid.

[0098] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1:0.8.

[0099] Example 11 This embodiment describes the process of adding excipients to prepare a pharmaceutical preparation.

[0100] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1:0.1.

[0101] The agent prepared using the reagent that inhibits the proliferation of pancreatic cancer cells is made by adding 100 times the amount of water to the reagent that inhibits the proliferation of pancreatic cancer cells, and adding 0.02 times the amount of phospholipid emulsifier, and mixing them to obtain an emulsion.

[0102] Example 10 This embodiment describes the process of adding excipients to prepare a pharmaceutical preparation.

[0103] An inhibitory agent for the proliferation of pancreatic cancer cells is composed of rapamycin, isopropyl myristate, and levulinic acid in a mass ratio of 10:1:0.1.

[0104] The agent prepared using the reagent that inhibits the proliferation of pancreatic cancer cells is made by adding 100 times the amount of starch to the reagent that inhibits the proliferation of pancreatic cancer cells to obtain a powder.

[0105] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. If such modifications and variations fall within the scope of equivalents of this invention, then this invention also intends to include these modifications and variations.

Claims

1. A reagent for inhibiting the proliferation of pancreatic cancer cells, characterized in that, It is composed of rapamycin, isopropyl myristate and levulinic acid in a mass ratio of 10:1~2:0.1~1.

2. The reagent for inhibiting the proliferation of pancreatic cancer cells according to claim 1, characterized in that, The pancreatic cancer cells are MIA PaCa-2 and / or PANC-1.

3. The application of the reagent for inhibiting the proliferation of pancreatic cancer cells according to claim 1, characterized in that, Drugs used to treat pancreatic cancer.

4. The application according to claim 3, characterized in that, The drug has an inhibitory agent that inhibits the proliferation of pancreatic cancer cells as its sole active ingredient.

5. The application according to claim 3, characterized in that, The active ingredient of the drug is a mixture of an agent for the proliferation of pancreatic cancer cells and a known anti-pancreatic cancer drug in equal mass proportions.

6. The application according to claim 5, characterized in that, The known anti-pancreatic cancer drugs are at least one of gemcitabine, albumin-bound paclitaxel, capecitabine, irinotecan, oxaliplatin, erlotinib, and olaparib.

7. The application according to claim 3, characterized in that, The drug contains pharmaceutically acceptable excipients.

8. The application according to claim 7, characterized in that, The pharmaceutically acceptable excipients include at least one of the following: fat-soluble solvents, co-solvents, oily carriers, solubilizers, emulsifiers, dispersants, binders, diluents, adhesives, disintegrants, lubricants, and preservatives.

9. The application according to claim 7, characterized in that, The drug is in capsule form, and the appropriate pharmaceutically acceptable excipients are diluents, binders, disintegrants, lubricants, and preservatives. Alternatively, the drug may be in the form of a solid dosage form, and the appropriate pharmaceutically acceptable excipients may be dispersants and binders. Alternatively, the drug may be in the form of a liquid, and the appropriate excipients may be emulsifiers, solubilizers, dispersants, lipid-soluble solvents, and co-solvents.

10. The application according to claim 9, characterized in that, The active ingredient in the drug has a mass percentage of 10 mg / g to 50 mg / g.

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