CN103435573B

CN103435573B - Benzyl-substituted thiazolocyclohexane compounds, and preparation methods and antitumor uses thereof

Info

Publication number: CN103435573B
Application number: CN201310308100.6A
Authority: CN
Inventors: 郭章华
Original assignee: Zhejiang Medical College
Current assignee: Zhejiang University ZJU
Priority date: 2013-07-16
Filing date: 2013-07-16
Publication date: 2015-04-01
Anticipated expiration: 2033-07-16
Also published as: CN103435573A

Abstract

The invention relates to the anticancer related medicine field, and concretely relates to thiazolocyclohexane compounds having a structure represented by general formula I and having an antitumor performance, and preparation methods, medicinal compositions and uses thereof. All groups in the formula I are defined in the specification.

Description

Benzyl-substituted thiazolocyclohexane compounds, preparation method and application thereof

Technical Field

The invention relates to the field of anti-tumor related medicaments. In particular, the invention relates to thiazolocyclohexane derivatives with anti-tumor effect, a preparation method thereof, a pharmaceutical composition containing the same and application thereof in the aspect of anti-tumor.

Background

Cancer is the leading disease threatening human life, and according to statistics, the total number of cancer deaths in the world reaches 700 million people every year, and more than 100 million patients who die of tumor in China every year are gradually increased, and the cancer is the first cause of death of urban population. At present, a plurality of traditional medicines for treating cancer diseases are clinically available in China, and the traditional medicines have obvious clinical treatment effects, but have the following defects: low specificity and poor selectivity, causes obvious toxic and side effects, and is easy to generate serious multi-drug resistance phenomena of cancers.

With the development of molecular biology, the current anti-cancer drugs are developing from traditional cytotoxic drugs to novel anti-cancer drugs aiming at multi-link mechanism, and one of the important new targets of the current anti-cancer effect concerned at home and abroad is protein tyrosine kinase (new targets of xanthone, butyl-Jian and anti-tumor drugs, Chinese prescription drugs, 2006, 12(57), 10-15). More than 20 receptor tyrosine kinases and non-receptor tyrosine kinases belonging to different families are currently used as targets for screening anticancer drugs, inhibitors of the protein tyrosine kinases have been on the market, and in order to find drugs with better activity, molecular targeted anticancer drug therapy also provides another challenging concept in recent years: the strategy of multiple target tyrosine kinase inhibition (multiple targeted tyrosine kinase inhibition) is an important direction for anti-tumor.

The invention discloses a protein tyrosine kinase inhibitor containing a thiazolocyclohexane structure, which can be used for preparing antitumor drugs.

Disclosure of Invention

The invention aims to provide a protein tyrosine kinase inhibitor containing a thiazolocyclohexane structure with a general formula I and pharmaceutically acceptable salts thereof.

It is another object of the present invention to provide a process for the preparation of compounds having the general formula I and pharmaceutically acceptable salts thereof.

It is a further object of the present invention to provide pharmaceutical compositions containing compounds of formula I and pharmaceutically acceptable salts thereof as active ingredients, together with one or more pharmaceutically acceptable carriers, excipients or diluents, and their use in anti-tumor applications.

The present disclosure will now be described in detail for the purpose of the invention.

The compounds of the present invention having the general formula I have the following structural formula:

wherein,

r is selected from H, hydroxyl and sulfydryl.

Preferably the following compound, or a pharmaceutically acceptable salt thereof, is selected from:

the compound of the general formula I is synthesized by the following steps:

compound a and compound B are reacted in the presence of a base in a suitable solvent to provide compound I.

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

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

The compound of the general formula I has an inhibiting effect on cancers, and can be used as an effective component for preparing a medicament for treating cancers. The activity of the compound of the general formula I is verified by an in vitro anti-tumor model.

The compounds of formula I of the present invention are effective over a relatively wide dosage range. The actual dosage of the compounds of formula I to be administered according to the invention can be determined by the physician in the light of the relevant circumstances. These include: the physical state of the subject, the route of administration, the age, body weight, individual response to the drug, severity of the symptoms, and the like.

Detailed Description

The present invention will be further described with reference to the following examples. It should be noted that the following examples are only for illustration and are not intended to limit the present invention. Variations of the teachings of the present invention may be made by those skilled in the art without departing from the scope of the claims of the present application.

Example 1

Reaction raw materials: commercially available or self-made.

1.67g (10mmol) of Compound A-1 and 2.75g (10mmol) of Compound B-1 were dissolved in 20mL of dry THF, and 3.04g (30mmol) of Et was added₃N, then stirred at room temperature until the reaction is complete (24-48 hours). Pouring the reaction mixture into ice water, extracting with 50mL of multiplied by 3 dichloromethane, combining organic phases, washing with a salt solution, drying with anhydrous sodium sulfate, evaporating the solvent on a rotary evaporator to obtain a crude product of the I-1, and purifying by column chromatography to obtain a pure product of the I-1, a white solid, mp.185-187 ℃; MS, M/z 362([ M + H)]⁺)。

Example 2

1.67g (10mmol) of Compound A-1 and 2.75g (10mmol) of Compound B-1 were dissolved in 20mL of dry THF, 3.88g (30mmol) of DIPEA was added, followed by refluxing with stirring until the reaction was complete (within 10 hours). The reaction mixture was poured onto iceExtracting with 50mL of dichloromethane with the concentration of 50X 3 in water, combining organic phases, washing with saline solution, drying with anhydrous sodium sulfate, evaporating the solvent on a rotary evaporator to obtain a crude product of I-1, and purifying by column chromatography to obtain a pure product of I-1, a white solid, mp.185-187 ℃; MS, M/z 362([ M + H)]⁺)。

Example 3

1.51g (10mmol) of Compound A-1 and 2.75g (10mmol) of Compound B-1 were dissolved in 20mL of dry THF, 3.88g (30mmol) of DIPEA was added, followed by refluxing with stirring until the reaction was complete (within 10 hours). Pouring the reaction mixture into ice water, extracting with 50mL of dichloromethane multiplied by 3, combining organic phases, washing with salt solution, drying with anhydrous sodium sulfate, evaporating the solvent on a rotary evaporator to obtain a crude product of the I-2, and purifying by column chromatography to obtain a pure product of the I-2, a white solid, mp.216-219 ℃; MS, M/z 346([ M + H)]⁺)。

Example 4

1.67g (10mmol) of Compound A-1 and 2.75g (10mmol) of Compound B-1 were dissolved in 20mL of dry THF, 3.88g (30mmol) of DIPEA was added, followed by refluxing with stirring until the reaction was complete (within 10 hours). Pouring the reaction mixture into ice water, extracting with 50mL of dichloromethane multiplied by 3, combining organic phases, washing with salt solution, drying with anhydrous sodium sulfate, evaporating the solvent on a rotary evaporator to obtain a crude product of I-3, and purifying by column chromatography to obtain a pure product of I-3, a white solid, mp.169-171 ℃; MS, M/z 362([ M + H)]⁺)。

Example 5

(1) Material

Cell lines: leukemia HL-60 cells, gastric adenocarcinoma SGC-7901 cells, breast cancer MCF-7 cells and lung cancer A-549 cells, which are all purchased from Shanghai cell research institute of Chinese academy of sciences.

Reagent: subpackaging MTT and Amresco; DMEM medium, Gibco; calf serum, lakehou marine life; packaging trypsin and Amresco; fluorouracil injection, 0.25g/10ml (Branch), Tianjin Jinyao amino acids, Inc.

The instrument comprises the following steps: clean bench, Suzhou clean plant; CO 2₂Incubator, Thermo corporation, model: HERA Cell 150; inverted microscope, Carl Zeiss, type: axiovert 200; enzyme linked immunosorbent assay, TECAN, type: sunrise; centrifuge, Kerdro corporation, model: heraeus.

(2) Method of producing a composite material

Cell culture: inoculating tumor cells in DMEM culture solution containing 10% calf serum, 100IU/ml penicillin G sodium salt and 100ug/ml streptomycin sulfate, standing at 37 deg.C and 100% relative humidity, and containing 5% CO₂The culture box of (5), passage 3 times for standby.

Determination by MTT method: cells in logarithmic growth phase are taken, digested by 0.25% trypsin (suspension cells are not required to be digested), suspended in DMEM culture solution containing 10% calf serum, slightly blown by a glass dropper to form single cell suspension, and counted by a blood cell counting plate under a microscope. The 96-well culture plate was inoculated with 90. mu.l of cell suspension per well (cell concentration was adjusted to 6 to 8X 10)⁴Pieces/ml) at 37 deg.C, 100% relative humidity, 5% CO₂After 24 hours of incubation in a 95% air incubator, 5. mu.l of the drug solution (final concentration: 10. mu.g/ml) was added to each well. In addition, each concentration was set with a negative control (equal concentration of DMSO) and a blank background (no cells added), and each group was set with 6 replicate wells. The culture was continued for another 48h, then 10. mu.l of 5mg/ml MTT solution was added to each well, and after further 4h, the supernatant (suspension of cells, required for suspension) was carefully aspiratedCentrifugation followed by aspiration of the supernatant). Add 100. mu.l DMSO into each well, shake in a micro-oscillator to dissolve the crystal completely, and test OD value by single-wavelength colorimetry with a 492nm microplate reader. The cell growth inhibition rate was calculated as an evaluation index in the following manner.

The inhibition ratio (%) was [1- (experimental OD mean-blank OD mean)/(control OD mean-blank OD mean) ] × 100%.

(3) Results

TABLE 1 inhibition ratio (%) of the samples to in vitro cultured tumor cells

(4) Conclusion

From the in vitro test results, the compound of the general formula I has strong inhibition effect on the 4 human cancer cells after being acted for 48 hours in vitro at the concentration of 10 mu g/ml.

Claims

1. The following compounds or pharmaceutically acceptable salts thereof:

2. a method of synthesizing the compound of claim 1, comprising the steps of:

wherein R is selected from H and hydroxyl and is para-substituted;

reacting the compound A and the compound B in a proper solvent in the presence of alkali to obtain a compound I;

3. Use of a compound of claim 1 for the preparation of an anti-tumor medicament.