CN103408541B - Indole-substituted thiazolo cyclohexane compound and antineoplastic applications thereof - Google Patents

Abstract

The present invention relates to the field of medicines related to anticancer. Specifically, the present invention relates to indole-substituted thiazolocyclohexane derivatives having the structure of formula I and having antitumor activity, as well as their preparation method, pharmaceutical composition and application.

Description

Indole-substituted thiazolocyclohexane compounds and their antitumor applications

technical field

The invention relates to the field of anti-tumor related drugs. Specifically, the present invention relates to thiazolocyclohexane derivatives with antitumor effects, their preparation methods, pharmaceutical compositions containing them and their antitumor application.

Background technique

Cancer is the primary disease that threatens human life. According to statistics, the total number of cancer deaths in the world reaches 7 million every year. In my country, more than 1 million patients die of tumors every year, and gradually increase. It has become the first cause of death in urban population. At present, there are many traditional drugs for the treatment of cancer in our country, and they have obvious therapeutic effects in clinical practice, but their disadvantages are: low specificity and poor selectivity, leading to obvious toxic and side effects, and prone to serious cancer multidrug resistance drug phenomenon.

With the development of molecular biology, today's anti-cancer drugs are developing from traditional cytotoxic drugs to new anti-cancer drugs that target the multi-link mechanism of the mechanism. It is one of the most important new targets of anti-cancer effects at home and abroad. It is protein tyrosine kinase (Huang Min, Ding Jian, New Targets of Antineoplastic Drugs, "Chinese Prescription Drugs", 2006, 12(57), 10-15). Protein tyrosine kinase At present, more than 20 receptor and non-receptor tyrosine kinases belonging to different families are used as targets for anticancer drug screening, and several of their inhibitors are already on the market. In order to find drugs with better activity In recent years, another challenging concept has been put forward in molecular targeted anticancer drug therapy: the strategy of multiple targeted tyrosine kinase inhibition is an important direction of anti-tumor.

The invention discloses a protein tyrosine kinase inhibitor containing an indolyl-substituted thiazolocyclohexane structure, which has obvious inhibitory effect on tumor cells.

Contents of the invention

An object of the present invention is to provide a protein tyrosine kinase inhibitor having a formula I containing a thiazolocyclohexane structure and a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide methods for the preparation of compounds of formula I and pharmaceutically acceptable salts thereof.

Another object of the present invention is to provide a pharmaceutical composition containing a compound of formula I and a pharmaceutically acceptable salt thereof as an active ingredient, and one or more pharmaceutically acceptable carriers, excipients or diluents, and its application in antitumor.

The content of the present invention will now be specifically described in conjunction with the purpose of the present invention.

Compounds of the present invention having formula I have the following structural formula:

Formula I compound of the present invention is synthesized through the following steps:

Compound I can be obtained by reacting compound A and compound B in a suitable solvent in the presence of a base.

Wherein the base is selected from triethylamine, diisopropylethylamine, potassium carbonate, sodium carbonate, and the reaction temperature is from room temperature to the temperature at which the used solvent refluxes.

The pharmaceutically acceptable salts of the compound of formula I of the present invention include, but are not limited to, various inorganic acids, such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, etc., or organic acids, such as formic acid, acetic acid, citric acid, oxalic acid, Pharmaceutically acceptable salts formed from fumaric acid, maleic acid, amino acids, etc.

The compound of formula I in the present invention has an inhibitory effect on cancer, and can be used as an active ingredient to prepare a medicine for treating cancer. The activity of the compound of formula I described in the present invention is verified by an in vitro anti-tumor model.

The compounds of formula I according to the invention are effective over a fairly wide dosage range. The actual dose of the compound of formula I of the present invention can be determined by a doctor according to the relevant conditions. These conditions include: the physical state of the person being treated, the route of administration, age, weight, individual response to the drug, and the severity of symptoms.

Detailed ways

The present invention will be further described below in conjunction with embodiment. It should be noted that the following examples are only for illustration, but not for limiting the present invention. Various changes made by those skilled in the art according to the teaching of the present invention shall be within the scope of protection required by the claims of the present application.

Example 1

Reaction raw materials: commercially available or self-made.

1.90 g (10 mmol) of compound A and 2.75 g (10 mmol) of compound B were dissolved in 20 mL of dry THF, and 3.04 g (30 mmol) of Et ₃ N were added, followed by stirring at room temperature until the reaction was complete (24-48 hours). The reaction mixture was poured into ice water, extracted with 50 mL×3 dichloromethane, the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and the solvent was evaporated on a rotary evaporator to obtain a crude product of I, which was purified by column chromatography. The pure product of I was obtained as a white solid, mp.157-159°C; MS, m/z=385 ([M+H] ⁺ ).

Example 2

1.90 (10 mmol) of compound A and 2.75 g (10 mmol) of compound B were dissolved in 20 mL of dry THF, and 3.88 g (30 mmol) of DIPEA were added, followed by reflux with stirring until the reaction was complete (within 10 hours). The reaction mixture was poured into ice water, extracted with 50 mL×3 dichloromethane, the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and the solvent was evaporated on a rotary evaporator to obtain a crude product of I, which was purified by column chromatography. The pure product of I was obtained as a white solid, mp.157-159°C; MS, m/z=385 ([M+H] ⁺ ).

Example 3

(1) Material

Cell lines: leukemia HL-60 cells, gastric adenocarcinoma SGC-7901 cells, breast cancer MCF-7 cells, and lung cancer A-549 cells were all purchased from Shanghai Cell Institute, Chinese Academy of Sciences.

Reagents: MTT, Amresco aliquots; DMEM medium, Gibco; calf serum, Lanzhou Minhai Biology; trypsin, Amresco aliquots; fluorouracil injection, 0.25g/10ml (branch), Tianjin Jinyao Amino Acid Co., Ltd.

Instruments: ultra-clean bench, Suzhou Purification Equipment Factory; _CO2 incubator, Thermo Company, model: HERA Cell150; inverted microscope, Carl Zeiss Company, model: Axiovert200; enzyme-linked immunoassay instrument, TECAN Company, model: Sunrise; centrifugation Machine, Kerdro Corporation, model: Heraeus.

(2) method

Cell culture: Tumor cells were inoculated in DMEM culture medium containing 10% calf serum, 100IU/ml penicillin G sodium salt and 100ug/ml streptomycin sulfate, placed at 37°C, 100% relative humidity, and 5% _CO2 In the incubator, after 3 passages, it is ready for use.

Determination of MTT method: Take the cells in the logarithmic growth phase, digest them with 0.25% trypsin (the suspension cells do not need to be digested), suspend them in DMEM culture medium containing 10% calf serum, and gently blow them into single cells with a glass dropper. For the cell suspension, live cells were counted with a hemocytometer under a microscope. Inoculate 90 μl of cell suspension in each well of a 96-well culture plate (adjust the cell concentration to 6-8×10 ⁴ cells/ml), and culture in an incubator at 37°C, 100% relative humidity, 5% CO ₂ , and 95% air for 24 hours After that, add 5 μl of drug solution to each well (final concentration is 10 μg/ml). In addition, a negative control (equal concentration of DMSO) and a blank background (without adding cells) were set for each concentration, and 6 replicate wells were set for each group. Continue culturing for another 48 hours, then add 10 μl of 5 mg/ml MTT solution to each well, continue culturing for 4 hours, and then carefully suck off the supernatant (suspended cells need to be centrifuged first, and then suck off the supernatant). Add 100 μl DMSO to each well, shake with a micro-oscillator to completely dissolve the crystals, and measure the OD value with a single-wavelength microplate reader at 492 nm. The cell growth inhibition rate was calculated by the following method as an evaluation index.

Inhibition rate (%)=[1-(mean OD value of experimental group-mean OD value of blank group)/(mean OD value of control group-mean OD value of blank group)]×100%.

(3) Results

Table 1. Inhibition rate (%) of samples to tumor cells cultured in vitro

(4 Conclusion

It can be seen from the above in vitro test results that the compound of formula I of the present invention has a strong inhibitory effect on these four human cancer cells after being acted in vitro for 48 hours at a concentration of 10 μg/ml.

Claims (2)

1. there is compound and the pharmaceutically acceptable salt thereof of formula I structure:

2. the formula I that claim 1 defines is preparing the purposes in antitumor drug.