CN106632000A - Application of diaryl ketone compounds in preparation of antitumor drug - Google Patents

Abstract

The invention relates to the field of antitumor drugs and provides application of diaryl ketone compounds in preparation of the antitumor drug. The diaryl ketone compounds are compounds shown as formulas M11, M13, M14, M15, M-126, M128, M130, M131, M132 and M149. By antitumor activity research experiments, the compounds show anti-tumor cell proliferation activity with a certain degree, and most compounds show strong inhibition effect; the compounds can strongly inhibit the growth of tumor cells, can promote apoptosis at relatively-low concentration and show stronger inhibition effect compared with positive contrast compounds. The preparation process of the compounds is simple and feasible; raw materials are low in price and easy to obtain; an antitumor effect target of the compounds can be found by researching a structure-function relationship, and a valuable guiding function is provided for further research and development of the drugs. The diaryl ketone compounds disclosed by the invention provide a basis for screening new drugs.

Description

Application of diaryl ketone compounds in the preparation of antitumor drugs

technical field

The invention belongs to the field of antitumor drugs, and in particular relates to the application of diaryl ketone compounds in the preparation of antitumor drugs.

Background technique

Malignant tumor is a common and frequently-occurring disease that seriously threatens human health. The mortality rate of human beings caused by malignant tumors ranks second among all diseases, becoming the second "killer" after cardiovascular diseases. At the beginning of 2013, the National Cancer Registry Center issued a version of the "2012 China Cancer Registration Annual Report", indicating that there are about 3.12 million new cancer cases in China every year, and more than 2 million cancer deaths, which means that 6 people are diagnosed with cancer every minute. cancer. At present, the incidence of cancer in the whole country is severe, and the incidence and mortality are on the rise. Cancer has become one of the major problems that seriously threaten the safety of human life and affect the quality of human life, and the problems caused by it have increasingly become a burden on social and economic development.

At present, tumor treatment methods mainly include surgery, radiation therapy and drug therapy (chemotherapy), but drug therapy is still the main method to a large extent. Most common solid tumors such as lung cancer, liver cancer, colon cancer, and pancreatic cancer still lack effective drugs, and many anticancer drugs have developed drug resistance during clinical application. With the advancement of society's science and technology, human beings' understanding of cancer has gradually become clearer, and the methods, technologies and theories for the prevention and early diagnosis of certain cancers have also become more mature, but they cannot completely cure cancer, and can only prolong the life of cancer in a very limited way. patient survival time. Therefore, the research and development of new anti-tumor drugs is imperative, and the continuous development of new anti-tumor drugs with high efficiency and low toxicity has become a research work of great significance.

Diaryl ketone compounds are a class of important additives and chemical intermediates. Back in the 1950s. Diaryl ketone compounds have been widely used in developed industrialized countries such as Europe and America. Mainly used in plastics, medicine, spices and electronics industries and other fields. In terms of pharmaceutical products, diaryl ketone compounds are intermediates of the antihistamine perhexiline, the narcolepsy drug modafinil, and the new powerful cerebral vasodilator flunarizine. In terms of spices, because it has the scent of roses, it is often used as a perfume fixative, and it is also used as a raw material for many perfumes and soap essences. The development and application of diaryl ketone compounds in my country started relatively late. It was not until the 1990s that diaryl ketone plants were built and put into production. The applicant's project team independently synthesized a series of diaryl ketone compounds, and disclosed the preparation methods of these compounds in the journal Tetrahedron Letters in 2016. Its biological activity has not been evaluated.

Contents of the invention

The object of the present invention is to aim at the above-mentioned present situation, provide the application of a series of diaryl ketone compounds and pharmaceutically acceptable salts thereof in the preparation of antitumor drugs, said diaryl ketone compounds are one of the following formulas:

Further, the pharmaceutically acceptable salts of diaryl ketone compounds provided by the present invention are hydrochloride, sulfate, phosphate and organic acid salts, such as methanesulfonate, trifluoromethanesulfonate, acetic acid Salt, Trifluoroacetate, Benzoate.

Further, the diaryl ketone compound is preferably of formula M11, M13, M15, M126, M131, M132, and the drug is used for the treatment of liver cancer, lung cancer, breast cancer, doxorubicin-resistant breast cancer, gastric cancer , cervical cancer, and glioma drugs.

Further, the medicine is a medicine for treating cervical cancer.

Further, the diaryl ketone compounds are preferably formula M131, M132, and the drug is used for the treatment of liver cancer, lung cancer, breast cancer, doxorubicin-resistant breast cancer, gastric cancer, cervical cancer, glioma Drug.

Further, the diaryl ketone compound is preferably M131, M132, and the drug is a drug for treating doxorubicin-resistant breast cancer.

The diaryl ketone compound and the pharmaceutically acceptable salt thereof of the present invention are used as an active ingredient and a conventional pharmaceutical carrier to prepare a pharmaceutical composition.

The diaryl ketone compounds of the present invention and pharmaceutically acceptable salts thereof can be prepared into various practical medicaments, such as granules, tablets, pills, capsules, injections, suspensions or emulsions, by conventional methods.

The present invention proves that the diaryl ketone compound has a good MCF-7/ADR tumor cells that are resistant to the positive control drugs 5-Fu and DOX also exhibit strong inhibitory effects.

The preparation of diaryl ketones involved in the present invention refers to the method of literature Tetrahedron Letters2016, 57, 90-94, specifically using transition metal palladium as a catalyst and phenylacetylene as an acylating agent to induce Under the action of aroyl ring, aroylation is carried out at the ortho position of the aromatic ring to obtain the final acylated product.

In view of the simple and easy preparation process of this type of compound, the raw material is cheap and easy to obtain; the series of compounds can be used to find their anti-tumor targets through the study of structure-activity relationship, and provide valuable guidance for further drug development. Therefore, the diaryl ketone compound of the present invention provides a basis for new drug screening.

Description of drawings

Figure 1 is a series of inhibitory activities of diaryl ketone compounds (40 μg/mL) on different tumor cells.

Fig. 2 Lesion effect of tumor cells HepG2, Hela, C6, MCF-7, MCF-7/ADR induced by M131 (20 μg/mL) and M15 (40 μg/mL).

Figure 3 Compounds M131, M15, and 5-Fu concentration-dependently inhibit the activity of tumor cells HepG2, Hela, and C6.

Fig. 4 Apoptosis detection diagram of tumor cell Hela induced by compound 5-Fu (100 μg/mL), M131 (10 μg/mL), M15 (20 μg/mL).

detailed description

The features and advantages of the present invention can be further understood through the following detailed description in conjunction with the accompanying drawings. The examples provided are only illustrative of the method of the present invention and do not limit the rest of the present disclosure in any way.

Hereinafter, unless otherwise specified, the materials and operating methods used in the present invention are well known in the art. [Example 1] In vitro anti-tumor activity evaluation of compounds

Tested tumor cells: human liver cancer cells HepG2, human lung cancer cells A549, human breast cancer cells MCF-7, human breast cancer cells resistant to doxorubicin (MCF-7/ADR), human gastric cancer cells SGC-7901, human cervical cancer cells Cell Hela, human glioma cell C6.

Cell culture: GIBCO DMEM medium, 10% fetal bovine serum and 0.01% L-glutamine were prepared as culture medium. The cultured cell lines were routinely cultured and passaged at 37°C and 5% CO ₂ saturated humidity, and cells in the logarithmic growth phase were used in the experiments.

In vitro anti-tumor activity evaluation (MTT method):

The above tumor cells were respectively plated on 96-well plates, and after being cultured in a 5% CO2 incubator at 37°C to cover a single layer, the cell culture medium was discarded, and cell maintenance medium containing different concentrations of test compounds were added to continue the culture, so as to avoid drug-free tumor cells. The affected cells were used as a blank control, and the antineoplastic drugs 5-fluorouracil (5-Fu) and doxorubicin (DOX) were used as positive controls. Eight replicate wells were set up in each group, and the culture was continued for 48 hours and then 72 hours. Visually inspect and record the cell condition with a microscope, add 30 μl of MTT (5 mg/mL) to each well and continue to incubate for 4 hours, discard the supernatant, add 30 μl DMSO to each well, incubate at 37°C for 10 minutes, and detect the absorbance at 492nm wavelength (A492) with a microplate reader . Calculate the average inhibition rate according to the following formula:

Inhibition rate=(average OD492 value of cell control group-average OD492 value of drug group)/average OD492 value of cell control group)×100%.

The test results show that all the above-mentioned diaryl ketone compounds have strong inhibitory activity on the proliferation of Hela, C6, HepG2, SGC-7901, A549, MCF-7, MCF-7/ADR tumor cells, and their half inhibitory concentration IC50 As shown in Table 1 below. Among them, compounds M11, M13, M15, M-126, M131, and M132 had the strongest inhibitory effects, and the average IC50 for seven tested tumor cells was ≤20 μg/mL.

For MCF-7/ADR tumor cells, not only the control drug DOX showed drug resistance, but another control drug 5-Fu also showed drug resistance. The half inhibitory concentration (IC50) of the cells was over 100μg/mL. However, the series of diaryl ketone compounds synthesized by us still have a good inhibitory effect on MCF-7/ADR tumor cells, among which the most active compounds M131 and M132 have IC50 of 3.9 μg/mL and 2 μg/mL, respectively. In addition, DOX has a strong inhibitory effect on the other six tumor cells, with an average IC50 of 2.5 μg/mL, compared with M131 and M132 (the average IC50 of the seven tumor cells is 5.7 μg/mL and 5.2 μg/mL) At a comparable level, it is superior to other compounds. While 5-Fu has an average IC50 of 31.1 μg/mL for the other six tumor cells, and our synthetic compound M-128 μg/mL (average IC50 is 31.1 μg/mL), M130 (average IC50 is 39.9 μg/mL), M149 (average IC50 is 34.5μg/mL), M14 (average IC50 is 25.6μg/mL) at the same level of inhibition, in addition to the other compounds we synthesized have stronger inhibitory effect on various tumor cells than 5-Fu .

Table 1 series of diaryl ketone compounds for the inhibitory activity of various tumor cells

In Table 1: a IC50: half inhibitory concentration. IC50<20μg/mL is recorded as +++, IC5020～50μg/mL is recorded as ++, IC50>50μg/mL is recorded as +. b Dox, doxorubicin, positive control; c 5-Fu, 5-fluorouracil, positive control.

The inhibitory activity of 40 μg/mL series of diaryl ketone compounds on different tumor cells is shown in Figure 1. At 40μg/mL, it showed a strong inhibitory effect on all tumor cells, and the inhibitory effect on A549 cells was relatively poor. Among all the compounds, M131 and M132 showed the strongest antitumor activity. The positive control drug 5-Fu has a relatively low inhibitory rate on various tumor cells, while another positive control drug DOX has a super potent inhibitory effect on all other tumor cells except for MCF-7/ADR. inhibitory activity.

The concentration-dependent inhibitory activity of representative compounds M131, M15 and the positive control drug 5-Fu on different tumor cells is shown in Figure 2. M131 and M15 exhibit concentration-dependent inhibitory activity on all tumor cells, and can achieve the effect of completely apoptotic and lethal tumor cells under the condition of sufficient concentration. The control drug 5-Fu showed a weak concentration dependence, and when the overall concentration was 5 to 40 μg/mL, the inhibitory activity on tumor cells did not increase significantly, showing a relatively uniform inhibitory effect.

[Example 2] The tumor cell pathological effect caused by the compound

We further photographed and recorded the cytopathic effects of Hela, C6, HepG2, MCF-7, MCF-7/ADR tumor cells induced by representative compounds M131 and M15. The specific implementation method is as follows:

Hela, C6, HepG2, MCF-7, MCF-7/ADR tumor cells in the logarithmic growth phase were respectively plated on 24-well plates, cultured in a 37°C, 5% CO2 incubator to cover the monolayer, and the cell culture medium was discarded , adding 20 μg/mL M131 and 40 μg/mL M15 test compound cell maintenance solution respectively to continue culturing. At 48 hours, the cells were visually observed under a microscope and photographed for cell lesions.

The tumor cell pathological effect induced by the compound is shown in Figure 3. Under the microscope, the untreated tumor cells grow well, adhere firmly to the wall, plump in shape, and have clear borders. 20μg/mL M131, 40μg/mL M15 treatment for 48h can lead to apoptosis of all tumor cells, the cells become round in different forms, and fall off from the culture plate. It can be seen that M131 and M15 have strong growth inhibitory effects on various tumor cells.

[Example 3] Evaluation of tumor cell apoptosis activity caused by compounds

The applicant further implemented the compound-induced tumor cell apoptosis test. The test conditions are as follows: Hela cells in the logarithmic growth phase were plated in a 24-well plate, and after the monolayer was covered, they were added with 10 μg/mL M131, 20 μg/mL M15, 100 μg/mL M15, and 100 μg/mL positive The control drug 5-Fu was incubated in the cell maintenance solution, and the cell control without drug treatment was set. After about 48 hours, the cells were collected, and the apoptosis was performed on the flow cytometer by using the Annexin V-FITC/PI apoptosis detection kit detection.

The experimental results shown in Figure 4 indicate that 10 μg/mL M131 and 20 μg/mL M15 can strongly induce the apoptosis of the above-mentioned tumor cells tested. In Hela cells, the apoptotic rate of untreated positive control cells was 3.2%, and the treatment of M131 and M15 for 48 hours promoted 98.2% and 88.8% of the cells to undergo apoptosis, respectively, while the Hela cells treated with the positive control drug 5-Fu apoptotic The rate is only 71.3%.

In summary, the series of diaryl ketone compounds in the present invention have a good inhibitory effect on the proliferation of different tumor cells, and the MCF-7/ADR tumor cells that are resistant to the positive control drugs 5-Fu and DOX are also A strong inhibitory effect is exhibited, showing great application potential. If further clinical research is carried out, it is hoped that effective drugs against tumor diseases will be prepared.

Claims (9)

1. The application of diaryl ketone compounds and pharmaceutically acceptable salts thereof in the preparation of antitumor drugs, wherein the diaryl ketone compounds are one of the following formulas: 2. application according to claim 1, is characterized in that, the pharmaceutically acceptable salt of described diaryl ketone compound is hydrochloride, sulfate, phosphate and organic acid salt, as methanesulfonic acid Salt, Triflate, Acetate, Trifluoroacetate, Benzoate. 3. The application according to claim 1, wherein the tumor is a drug for liver cancer, lung cancer, breast cancer, doxorubicin-resistant breast cancer, gastric cancer, cervical cancer, and glioma. 4. The application according to claim 3, characterized in that the tumor is cervical cancer. 5. The application according to claim 1, wherein the diaryl ketone compound is any one of the formulas M11, M13, M15, M126, M131, M132. 6. The application according to claim 1, characterized in that, said diaryl ketone compound is of formula M131 or M132. 7. The application according to claim 6, characterized in that the drug is a drug for treating doxorubicin-resistant breast cancer. 8. A pharmaceutical composition, characterized in that it is made of the diaryl ketone compounds listed in claim 1 and pharmaceutically acceptable salts thereof as active ingredients and conventional pharmaceutical carriers. 9. The pharmaceutical composition according to claim 8, characterized in that it can be prepared into various practical medicaments, such as granules, tablets, pills, capsules, injections, suspensions or emulsions, by conventional methods.