CN103420993B - Compound containing thiophene substitution, its preparation method and use - Google Patents

Abstract

The invention relates to the field of medicines related to anticancer. Specifically, the present invention relates to a compound with general formula I having antitumor activity and its preparation method, pharmaceutical composition and use. Wherein the definition of each group is as described in the description.

Description

Compound containing thiophene substitution, its preparation method and use

technical field

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

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 people every year. In my country, more than 1 million patients die from cancer every year, and it is gradually increasing. 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 a pyrazole amide structure, which can be used to prepare antitumor drugs.

Contents of the invention

One object of the present invention is to provide a protein tyrosine kinase inhibitor having a general formula I containing a pyrazole amide structure.

Another object of the present invention is to provide the preparation of compounds of general formula I.

Another object of the present invention is to provide the application of the compound containing general formula I in anti-tumor.

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 general formula I have the following structural formula:

in,

R ₁ is selected from H, C1-C3 alkyl.

Preferred compounds of the present invention having general formula I are shown below:

The compound of general formula I 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, KOH, NaOH, potassium carbonate, sodium carbonate, sodium ethoxide, sodium hydride, and the solvent is selected from chloroform, dichloromethane, acetonitrile, DMF and the like. This reaction can use iodine salt as catalyst, such as KI and NaI etc.

The compound of the general 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 general formula I in the present invention is verified by anti-tumor model in vitro.

The compounds of general formula I according to the invention are effective over a fairly wide dosage range. The actual dosage of the compound of general formula I of the present invention can be determined by a doctor according to 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: self-made in laboratory, conventional method.

3.59g (10mmol) of compound A-1 and 2.16g (10mmol) of compound B-1 were dissolved in 20mL of dry DMF, then 4.15g (30mmol) of potassium carbonate and 0.50g of potassium iodide were added, and then heated to reflux overnight in a nitrogen atmosphere. The reaction mixture was poured into ice water, extracted with 50mL×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 the crude product of I-1, which was subjected to column chromatography After purification, the pure product of I-1 was obtained as a pale yellow solid, mp191-194°C, MS, m/z=512 ([M+NH ₄ ] ⁺ ).

Example 2-3

Referring to the operation of Examples 1 and 2, the compounds with formula I in the following table were synthesized.

Example 4

(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 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 incubate for 24 hours at 37°C, 100% relative humidity, 5% CO ₂ , and 95% air in an incubator 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 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

From the above in vitro test results, it can be seen that the compound of general formula I of the present invention has a strong inhibitory effect on these 4 kinds of human cancer cells after acting in vitro for 48 hours at a concentration of 10 μg/ml.

Claims (3)

1. Compounds with general formula I: in, R ₁ is selected from H, C1-C3 alkyl. 2. The compound of general formula I as defined in claim 1, selected from the group consisting of: 3. The use of the compound of general formula I as defined in claim 1 or 2 in the preparation of antitumor drugs.