Application of Curcuma oxadiol in the preparation of drugs for preventing, treating and inhibiting lung cancer

Abstract

The present invention discloses the use of zedoary oxadiol in the preparation of drugs for preventing, treating and inhibiting lung cancer, and relates to the technical field of traditional Chinese medicine. The present invention finds that zedoary oxadiol can effectively induce apoptosis of A549 cells, inhibit the growth of non-small cell lung cancer, and significantly inhibit the migration of A549 cells, hinder the development of non-small cell lung cancer, and inhibit the growth of tumors. The present invention provides a new use of zedoary oxadiol, expands the application of zedoary oxadiol in medicine, and provides a new technical means for the treatment of lung cancer.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicines, and in particular to the application of zedoary oxadiol in the preparation of drugs for preventing, treating and inhibiting lung cancer. Background Art

[0002] Lung cancer is the most common malignant tumor that poses the greatest threat to humans today. Lung cancer has the characteristics of high morbidity, high metastasis rate, and high mortality rate, with a 5-year survival rate of only 18%. Therefore, the diagnosis and treatment of lung cancer has become one of the research focuses and work centers of modern medical researchers. Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, accounting for about 85% of all lung cancer cases, and has become a top priority in prevention and treatment research. Non-small cell lung cancer includes different pathological subtypes such as squamous cell carcinoma, adenocarcinoma, large cell lung cancer, and poorly differentiated variants, but the treatment and prognosis methods used are basically similar. At this stage, surgical treatment is generally adopted as the main treatment, supplemented by chemoradiotherapy, targeted therapy, or immunotherapy. Although the above treatment methods can delay disease progression to a certain extent, there are serious side effects and drug resistance problems. Therefore, the need to develop new anti-tumor drugs is more urgent.

[0003] Traditional Chinese medicine has potential advantages in the field of anti-cancer due to its small side effects on the human body. Zedoary turmeric oil is a volatile oil extracted from the dried rhizome of Atractylodes lancea, containing ingredients such as curcumol, curdione, curcumenol, isocurcumenol, dehydrocurdione, curcuma diol, and curcumenone. Studies have found that it has antiviral, anti-inflammatory, anti-tumor, and antibacterial effects, and can also change and enhance the immunogenicity of tumor cells, and has a certain effect on the treatment of lung cancer.

[0004] Research from Guangdong Pharmaceutical University found that curcuma alcohol showed anti-tumor effects in both in vivo and in vitro experiments. For example, non-small cell lung cancer cells A549 and NCIH460 showed different degrees of proliferation inhibition after being treated with different concentrations of curcuma alcohol for different periods of time. This inhibition phenomenon was concentration-dependent and time-dependent. Through the exploration of its mechanism of action, it was believed that curcuma alcohol caused lung cancer cell apoptosis through the ROS / JNK signaling pathway. At the same time, curcuma alcohol could also arrest the cell cycle in the G / M phase, leading to cell proliferation inhibition.

[0005] Due to the large number of ingredients and unclear mechanism of action, Curcuma oil cannot maximize its therapeutic effect. At present, there are still very few monomeric Chinese herbal ingredients that have clear good therapeutic effects on lung cancer. It is urgent to discover more effective Chinese herbal monomers for the treatment of lung cancer and provide more effective options for the treatment of lung cancer.

[0006] Zedoarondiol (CAS No.98644-24-7) is an active ingredient in Curcuma zedoaria, and its chemical structure is as follows:

[0007]

[0008] Currently, its effect in treating lung cancer has not been reported. The inventors found in their research that Curcuma zedoariae oxadiol has good activity in treating lung cancer. In view of this, the present invention provides the use of Curcuma zedoariae oxadiol in the preparation of drugs for treating lung cancer with fewer side effects. Summary of the invention

[0009] The purpose of the present invention is to provide an application of zedoary oxadiol in the preparation of drugs for preventing, treating and inhibiting lung cancer, which has definite efficacy and small side effects.

[0010] To achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows:

[0011] The present invention provides application of zedoary oxadiol in preparing medicine for preventing, treating and inhibiting lung cancer.

[0012] Preferably, the lung cancer is lung cancer of mammals (including humans and non-human mammals).

[0013] Preferably, the lung cancer is at least one of small-cell lung cancer and non-small cell lung cancer.

[0014] More preferably, the lung cancer is non-small cell lung cancer.

[0015] Preferably, the lung cancer is at least one of lung adenocarcinoma, lung squamous cell carcinoma, large cell lung cancer, lung adenosquamous carcinoma, and salivary gland tumor.

[0016] Preferably, the prevention, treatment and inhibition of lung cancer is at least one of inhibiting the proliferation rate of lung cancer cells, changing the cell cycle distribution of lung cancer cells, promoting apoptosis of lung cancer cells, inhibiting the migration ability of lung cancer cells, and inhibiting the growth of lung cancer tissue.

[0017] Preferably, the prevention, treatment and inhibition of lung cancer is at least one of promoting apoptosis of lung cancer cells, inhibiting the migration ability of lung cancer cells and inhibiting the growth of lung cancer tissue.

[0018] Preferably, the prevention, treatment and inhibition of lung cancer is to simultaneously promote apoptosis of lung cancer cells, inhibit the migration ability of lung cancer cells and inhibit the growth of lung cancer tissue.

[0019] Preferably, in the application, Curcuma oxadiol can be used simultaneously with other anticancer agents.

[0020] Examples of the anticancer agent include acivicin, aclarubicin, acodazole hydrochloride, acronine, aldesleukin, altretamine, ambomycin, ametantrone acetate, aminoglutethimide, amsacrine, anastrozole, anthramycin, asparaginase, asperlin, azacitidine, azetepa, azotomycin, batimastat, benzodepa, bicalutamide, bisantrene hydrochloride, bizelesin, bleomycin sulfate, sulfate), brequinar sodium, bropirimine, calusterone, caracemide, carbetimer, carboplatin, carmustine, carubicin hydrochloride, carzelesin, cedefingol, chlorambucil, cirolemycin, fluorouracil, flurocitabine, fosquidone, iproplatin, irinotecan hydrochloride, lanreotide acetate, letrozole, liarozole hydrochloride, lometrexol sodium sodium), lomustine, losoxantrone hydrochloride, masoprocol, maytansine, mechlorethamine hydrochloridehydrochloride, megestrolacetate, melengestrol acetate, melphalan, menogaril, mitosper, mitotane, mitoxantrone hydrochloride, mycophenolic acid, nocodazole, nogalamycin, ormaplatin, oxisuran, paelitaxel, pegaspargase, peliomycin, pentamustine, peplomycin sulfate, perfosfamide, pipobroman, piposulfan, piroxantrone hydrochloride hydrochloride), plicamycin, plomestane, porfimer sodium, porfiromycin, prednimustine, procarbazine hydrochloride, puromycin, pyrazofurin, riboprine, rogletimide, safingol, safingol hydrochloride, semustine, simtrazene, sparlfosate sodium, sparsomycin, vapreotide, verteporfin, andrographolide, atrimustine, axinastatin 1, axinastatin 2 2) Marine cyclic peptide 3 (axinastatin3), azasetron, azatoxin, azatyrosine, batimastat, carboxyamidotriazole, cicaprost, dacliximab, ganirelix, lanreotide, lovastatin, loxoribine, lurtotecan, miltefosine, mofarotene, tirapazamine.

[0021] The beneficial effects of the present invention are:

[0022] (1) The present invention provides the use of curcuma oxadiol in the preparation of drugs for treating lung cancer, discovers a new use of curcuma oxadiol, expands the application of curcuma oxadiol in medicine, and provides a new technical means for the treatment of lung cancer;

[0023] (2) The experimental results of the present invention show that Curcuma zedoariae can effectively induce apoptosis of A549 cells, inhibit the growth of non-small cell lung cancer, and significantly inhibit the migration of A549 cells, hinder the development of non-small cell lung cancer, and inhibit tumor growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The figure is the result of the effect of Curcuma oxadiol on the viability of A549 cells in Example 1, wherein, compared with the blank group, ****p<0.0001, ***p<0.001, **p<0.01, *p<0.05.

[0025] Figure 2 This is a graph showing the effect of Curcuma zedoariae oxadiol on the wound healing rate of A549 in Example 2, wherein, compared with the blank group, **p<0.01, *p<0.05.

[0026] Figure 3 This is a graph showing the effects of various drugs in Example 3 on the tumor diameter of nude mice.

[0027] Figure 4 This is a graph showing the effects of various drugs in Example 3 on the body weight of tumor-bearing nude mice.

[0028] Figure 5 The results of the effects of the drugs in Example 3 on the spleen weight (spleen weight) of nude mice are shown in FIG. 3 , where **p<0.01 compared with the blank group; #p<0.05 compared with the model group.

[0029] Figure 6 The results of the effects of the drugs in Example 3 on the tumor weight of nude mice are shown in Figure 3, where compared with the blank group, **p<0.01; compared with the model group, #p<0.05, ##p<0.01.

[0030] Among them, “Control”, “con” and “CON” all refer to the blank group;

[0031] “Zedoarondiol” and “Zed” refer to the zedoarondiol-treated group;

[0032] “Model” refers to the model group;

[0033] “5-FU” refers to the positive drug treatment group;

[0034] “Cur” refers to the curcuminol-treated group;

[0035] “Zedoary Turmeric Oil” and “ZTO” both refer to the zedoary turmeric oil treatment group. DETAILED DESCRIPTION

[0036] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is only an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the invention of the present invention based on the disclosed content, and they should also fall within the scope of the present application.

[0037] The present invention is further described below by way of specific examples. The various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the operating methods used are conventional operating methods, the equipment used are conventional equipment, and the equipment materials used in each embodiment are the same. Unless otherwise specified, all percentages, ratios and proportions herein are by weight. Unless otherwise specified, all temperatures are in degrees Celsius (°C).

[0038] Zedoarondiol used in the present invention has a CAS number of 98644-24-7 and a chemical structural formula as follows:

[0039]

[0040] Example 1

[0041] 1. Materials

[0042] 1.1 Experimental cells

[0043] Human non-small cell lung cancer cells, A549 cells, were purchased from Wuhan Pronocell Biotechnology Co., Ltd.

[0044] 1.2 Drugs

[0045] Curcuma oxadiol is homemade with a purity of ≥98%.

[0046] 1.3 Reagents

[0047] Ham's F-12K medium (PM150910, Pronosai) and trypsin (0458) were from GIBCO; DMSO (D-5879, Sigma) and CCK8 (CK04, Dongren Chemical Technology Co., Ltd.) were from GIBCO.

[0048] 1.4 Instruments

[0049] MCO175 CO2 incubator (Sanyo); Sh1 inverted microscope (OLYMPUS); SYNERGYTM 4 microplate reader (BioTek).

[0050] 2. Methods

[0051] 2.1 Cell culture

[0052] The cells were revived and cultured in normal culture medium at 5% CO2 and 37°C. After two passages at a ratio of 1:3, the cells were collected and digested with trypsin to adjust the cell concentration to 1×10 5 / mL, and inoculated into 96-well culture plates, 6-well culture plates, 10 cm culture dishes, etc., and the medium was changed after 24 hours for subsequent experiments.

[0053] 2.2 Drug concentration determination

[0054] The experiment was divided into a normal group and different dose groups of Curcuma oxadiol. The drug doses were set as follows: 2mM, 1mM, 0.5mM, 0.25mM, 0.125mM, and 0.0625mM.

[0055] The cultured cells were washed 3 times with PBS, and Ham's F-12KD was added to the blank group, Ham's F-12KD and DMSO (added amount 0.1%) were added to the negative group, and Ham's F-12KD and drugs were added to the drug group. Before adding the drug, DMSO was used to dissolve it to a concentration of "mmol / mL", and then diluted 1:1000 with culture medium (Ham's F-12KD) to reach the target dose mM.

[0056] 2.3 Determination of cell viability

[0057] After the drug was applied for 24 h, 48 h, and 72 h, 100 µL of CCK8 solution was added to each well of the 96-well plate and incubated in an incubator for 2-4 h. The 96-well plate was placed on a shaker and shaken at a low speed for 10 min. The OD value was detected at a wavelength of 450 nm using an enzyme reader.

[0058] 2.4 Statistical methods

[0059] The quantitative data were expressed as mean and standard deviation. Western blot images were analyzed by Image lab for nonparametric tests. All statistical data were analyzed by GraphPad Prism software (San Diego, CA). One-way analysis of variance (ANOVA) was used to analyze multiple group comparisons. Student's t-test was performed to analyze intergroup comparisons. P values ​​< 0.05 were considered statistically significant.

[0060] The experimental results are shown in Figure 1 The results showed that Curcuma zedoariae oxadiol could significantly induce apoptosis of A549 cells at a concentration of 1 mM (P<0.05).

[0061] Example 2

[0062] 1. Materials (same as Example 1)

[0063] 2. Experimental method: A549 non-small cell lung cancer cells were cultured in a 6-well plate. After the cells grew to 5x105, a sterile 200μL pipette tip was used to gently draw a straight line on the culture dish. The culture dish was gently shaken, and the floating cells in the scratch area were washed off with culture medium. 2mM curcuma oxadiol was added to each well. A control group without curcuma oxadiol was set up. Before adding curcuma oxadiol for treatment, an inverted microscope was used to photograph and record the initial state of the scratch area; the cells were continued to be cultured in the culture medium containing curcuma oxadiol, and different treatment times (6 hours, 12 hours, and 24 hours) were selected. The state of the scratch area was photographed and recorded at each time point using an inverted microscope; by comparing the pictures before and after the experiment, the effect of curcuma oxadiol on the migration ability of A549 non-small cell lung cancer cells was evaluated.

[0064] Randomly draw 6 to 8 horizontal lines, calculate the mean value of the distance between cells or the mean value of the scratch area, and quantitatively analyze the inhibitory effect of Curcuma zedoariae oxadiol on cell migration ability.

[0065] Wound healing rate (cell migration rate) = (initial scratch area - scratch area at time t) / initial scratch area. Experimental results: The results are as follows Figure 2 As shown, Curcuma oxadiol can significantly inhibit the migration ability of A549 cells.

[0066] Example 3

[0067] Experimental methods:

[0068] Experimental animals: BALB / c-nu nude mice, body weight 20±2g.

[0069] Animal husbandry: All animals were placed in plastic cages in a stable environment with a 12 / 12 h light / dark cycle and free access to tap water and food. All animals were acclimated for 1 week at 25 °C and air humidity maintained at 40%-60%.

[0070] drug:

[0071] Curcuma oxadiol;

[0072] Positive drug 5-Fluorouracil (5-fluorouracil; 5-FU): 20 mg / kg (converted according to the instructions)

[0073] Curcumenol (C 15 H 22 O2, Cur) was purchased from Chengdu Purifa Co., Ltd., with the product number BP0420, 300 mg / kg, and the molecular formula is:

[0074]

[0075] Zedoary Turmeric Oil (ZTO) was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with the product number S27690, 240 mg / kg.

[0076] Grouping method: After 7 days of adaptive feeding, nude mice were randomly divided into groups. 5×10 6 A549 cells were cultured and the tumorigenesis was observed.

[0077] After tumor formation, animals in each treatment group were given corresponding doses of drugs for treatment. The administration method was oral administration, with an oral volume of 10 mL / kg, once a day, for 14 consecutive days. The model group and the normal control group were given an equal amount of normal saline in the same way. The animal weight and tumor diameter were measured once every 3 days during the whole process.

[0078] Experimental results: After nude mice developed tumors, they were given medication and the results were as follows Figure 3-6 As shown in the figure, compared with the model group, the tumor growth of the positive drug and each dose drug group was very slow, especially the positive drug group and the high dose drug group, while curcumenol and curcuma oil, which have similar structures to curcuma oxadiol, did not have significant anti-tumor effects.

[0079] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. Application of Curcuma zedoariae oxadiol in the preparation of drugs for inhibiting the migration ability of non-small cell lung cancer cells.

2. The use according to claim 1, characterized in that: The drug simultaneously inhibits the proliferation rate of non-small cell lung cancer cells, changes the cell cycle distribution of non-small cell lung cancer cells, promotes the apoptosis of non-small cell lung cancer cells, inhibits the migration ability of non-small cell lung cancer cells, and inhibits the growth of non-small cell lung cancer tissue.

3. The use according to claim 1, characterized in that: In the application, curcuma oxadiol and other anticancer agents are used simultaneously.

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