CN103709122B - Antitumor and the antifungal compound being used for the treatment of - Google Patents

Abstract

The present invention relates to medical technical field, preparation technology and the antitumor and antimycotic medical usage thereof that a class has antitumor activity and Antifungal Compounds, particularly, to cancer of the stomach, various metastatic hepatic carcinoma, breast cancer, lung cancer, colon cancer, the Several Kinds of Malignancies such as melanoma have obvious curative effects.

Description

Antineoplastic and antifungal compounds for therapy

technical field

The invention relates to the technical field of medicine, and relates to a preparation method of a fused tricyclic antitumor and antifungal compound used for treatment and an antitumor and antifungal medical application thereof.

Background technique

Malignant tumors seriously threaten human life and health. About 7 million people die from cancer every year in the world, accounting for about a quarter of the total death toll. Gastric cancer is the most common malignant tumor in my country, and its death rate accounts for the first place among all kinds of malignant tumors. Liver cancer is the second cancer "killer" in my country, and it has jumped to the first place in some high-incidence areas of liver cancer, accounting for 45% of liver cancer deaths worldwide. Breast cancer is the main malignant tumor that endangers women's health. About 1.2 million women in the world develop breast cancer every year. At present, the mortality rate of cancer patients in our country exceeds 30%, which has become the second largest factor in the death of Chinese residents. Drug therapy has become an effective and widely used treatment method for malignant tumors. In 2010, the global sales of antineoplastic drugs were about 60 billion US dollars. There are many kinds of antineoplastic drugs used clinically, among which chemotherapy drugs mainly include alkylating agent molybdenum complex antineoplastic drugs, anthracycline antineoplastic drugs, and DNA-damaging antibiotics. In addition, the research on natural anti-tumor drugs also occupies a considerable proportion. For example, some drugs commonly used in clinical practice include camptothecin, vincristine, and paclitaxel. However, the existing antitumor drugs have problems such as poor selectivity, toxic side effects, and drug resistance. Finding anti-tumor drugs with high efficiency and low toxicity is still an important issue for scientists. The invention provides a fused tricyclic drug structure with anti-tumor activity, and has important development and application prospects. In recent years, great progress has been made in the research of antifungal drugs, but there are still some deficiencies in the drugs currently used in clinical practice, such as: fluconazole has poor effect on Aspergillus, and drug-resistant strains are constantly emerging; The antibacterial effect is poor; itraconazole is not easy to pass through the blood-brain barrier, and the incidence of side effects is high. Therefore, finding highly effective, low toxicity, broad-spectrum antifungal drugs is still a worldwide important issue.

Contents of the invention

The present invention is based on a surprising discovery that fused tricyclic compounds or their pharmaceutically acceptable salts can effectively inhibit the growth of certain cancer cells, and such compounds have obvious inhibitory effects on gastric cancer, liver cancer, and breast cancer cells , In addition, the fused tricyclic compound or a pharmaceutically acceptable salt thereof also has an inhibitory effect on fungal growth.

Its structural formula is as follows:

X is selected from S, O, N-R', C=O

R ₁ and R' are selected from hydrogen, alkyl, R ₂ , R ₃ , R ₄ and R ₅ are respectively selected from mono-substituted and multi-substituted hydrogen, alkyl, cycloalkyl, alkoxy, aryl, hydroxyl, carboxyl, halogen, nitrile, nitro.

A subset of the fused tricyclic compounds have the formula:

The compounds exist in the form of hydrates or salts. Salts are inorganic salts or organic salts, such as hydrochloride, sulfate, fumarate, methanesulfonate, sulfonate, and the like.

Further, R ₁ is selected from hydrogen, alkyl, and benzyl, and R ₂ , R ₃ , R ₄ and R ₅ are each monosubstituted;

Furthermore, R ₂ , R ₃ , R ₄ and R ₅ are hydrogen, bromine, methoxy, chlorine, hydroxyl, carboxyl, and nitrile, respectively.

The term "alkyl" herein refers to straight or branched chain hydrocarbons containing 1 to 10 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl. The term "cycloalkyl" refers to saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 12 carbons. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclovinyl, cycloheptyl, and cyclooctyl. The term "alkoxy" refers to -O-alkyl. The term "aryl" refers to a 6-carbon monocyclic, 10-carbon bicyclic, 14-carbon tricyclic aromatic ring system, wherein each ring may have from 1 to 4 substituents. Examples of aromatic rings include, but are not limited to, phenyl, naphthyl, and anthracenyl. Alkyl, aryl, cycloalkyl, aryl, alkoxy referred to herein include substituted and unsubstituted moieties.

Some examples of fused tricyclic heterocyclic compounds of the present invention are shown below

Some other examples of compounds of the present invention are shown below:

The above-mentioned fused tricyclic compounds inhibit the growth of cancer cells and fungi. Accordingly, in another aspect, the invention also features methods of treating cancer and fungi. The method comprises administering to a subject in need thereof an effective one of the compounds described above.

detailed description

1: Synthesis of compound 1

The specific synthesis steps are: (0.5mmol), CuI (0.1mmol), L-proline (0.2mmol), and K2CO3 (2.5mmol) were added to the reactor, and the Ar gas was replaced three times. join in (0.55mmol) and 2.0mL DMSO, stirred at 90°C for 48h, continued to heat to 110°C for 48h, and monitored the reaction with TLC. The solvent was distilled off under reduced pressure, and the residue was dissolved in 20 mL of ethyl acetate. The organic phase was washed three times with water and once with brine, and dried over anhydrous sodium sulfate. Product 1 was obtained after concentration and purification by column chromatography. ¹ HNMR(400MHz,DMSO-d ₆ ):8.35(s,1H),7.14(dd,J=2.0,10.4Hz,1H),6.91(s,1H),6.65(s,1H),6.55(s, 1H),2.10(s,3H),2.09(s,3H); ¹³ CNMR(100MHz,DMSO-d ₆ )d148.9(J=243.8Hz),140.0,136.4,134.0,129.1(J=13.5Hz) ,124.7,123.8(J=9.4Hz),121.4(J=2.5Hz),120.7(J=3.2Hz),114.4(J=22.6Hz),114.1,111.8,20.5,19.1.MS(EI)m/z279 (M)+.

2: Synthesis of compound 19

Concrete synthetic steps are:

1.0 mmol of 2-iodophenol, 1.0 mmol of 1-F-2-nitrobenzene and 2 mmol of potassium carbonate were dissolved in 10 mL of DMSO, and heated at 100°C for 15 hours. After cooling to room temperature, a large amount of water was added and extracted three times with ethyl acetate. The organic phase was washed three times with water, concentrated and dried to obtain 2-(2-iodophenoxy)aniline. ¹ HNMR (400MHz, CDCl ₃ ): 7.85 (dd, J=1.5, 7.8Hz, 1H), 7.30 (dt, J=1.5Hz, J=7.8Hz, 1H), 6.92-6.82 (m, 3H), 6.67 (d,J=8.1Hz,2H),6.53(dd,J=1.5Hz,J=7.5Hz,1H),4.89(s,2H). ¹³ CNMR(100MHz,CDCl3):157.3,142.6,140.8,140.2 , 130.7, 126.0, 125.5, 120.5, 117.5, 117.3, 116.7, 88.7.

Mix 2-(2-iodophenoxy)aniline1mmol and 5mmol acetic anhydride, stir overnight at room temperature, add sodium carbonate aqueous solution to the mixture, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, concentrate and column chromatography to obtain the product N- (2-(2-iodophenoxy)phenyl)acetamid. ¹ HNMR(400MHz,CDCl3):8.36(d,J=7.9Hz,1H),7.94(s,1H),7.79(d,J=8.0Hz,1H) ,7.23(t,J=7.7Hz,1H),7.04(t,J=7.7Hz,1H),6.94(d,J=7.7Hz,1H),6.84(t,J=7.4Hz,1H),6.70 (d,J=8.1Hz,1H),2.11(s,3H). ¹³ CNMR(100MHz,CDCl3):δ=168.1,155.1,144.9,139.5,129.6,129.1,125.6,123.9,123.6,121.1,118.8, 116.9,88.2,24.5.(EI):m/z=353[M]+

Add 1mmol of N-(2-(2-iodophenoxy)phenyl)acetamid and 2mmol of potassium carbonate to the reactor, and under the protection of argon, add 0.1mmol of DMEDA and 1mL of toluene through a syringe. Stirring at 135°C for 24 hours, the reaction solution was cooled to room temperature, dissolved in dichloromethane, filtered, the solution was concentrated, and compound 19 was obtained by column chromatography. ¹ H NMR (400MHz, CDCl3): δ=7.50-7.45 (m, 2H), 7.22-7.16 (m, 2H), 7.15-7.10 (m, 4H), 2.33 (s, 3H). ¹³ CNMR (100MHz, CDCl3): δ=169.0, 150.8, 129.3, 126.7, 125.0, 123.2, 116.7, 23.0.

3: Synthesis of compound 25

2mmol, 2.0mmol, CuI0.2mmol, A mixture of 0.4mmol and Cs ₂ CO ₃ 4.0mmol was dissolved in 5mL of DMF, and heated to 120°C for 24 hours under the protection of N ₂ gas. After the reaction solution was cooled to room temperature, it was dissolved in ethyl acetate, filtered, washed with ethyl acetate, the filtrate was concentrated and dried, and purified by column chromatography to obtain compound 25.

4: Synthesis of compound 37

0.15mmol of 20mol% CuI was dissolved in a mixture of 0.4mL DMSO and 0.4mL chlorobenzene, and stirred at 140°C for 48 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, it was dissolved in ethyl acetate, concentrated to dryness, and purified by column chromatography to obtain compound 37. ¹ HNMR(400MHz,DMSO):δ6.05(s,2H),7.66(s,1H),7.93-8.01(m,2H),8.26-8.57(m,5H),8.87-8.95(m,2H) ,10.78(brs,1H,); ¹³ CNMR(100MHz,DMSO):54.34,122.60,123.18,125.43,125.72,126.36,130.05,131.23,134.00,135.72,138.19,138.72,145.53,2186m; [M+1] ⁺

The fused tricyclic compound of the present invention is a class of high-efficiency and low-toxicity anti-tumor active compounds, and has obvious curative effects on various malignant tumors such as various metastatic liver cancers, pancreatic tumors and melanomas. Its main efficacy test results are as follows:

1. Test method

Using a group of tumor cell lines, namely SK-Hep-1 (liver cancer), HepG2 (hepatocellular carcinoma), MKN-45 (gastric cancer), MDA-MB-231 (breast cancer cells), A549 (non-small cell lung cancer), OVCAR-3 (ovarian cancer), A498 (kidney cancer), HCT-116 (colon cancer), M4 (melanoma) to detect the in vitro antitumor activity of the above fused tricyclic compounds.

SK-Hep-1, HepG2, MKN-45, MDA-MB-231, A549, OVCAR-3, A498, HCT-116, M4 were kept in plastic dishes in PRMI medium filled with 5% fetal bovine serum. Seed tumor cells at a final density of 10,000 cells/mL in 96-well plates. Treat the cells with the test compound (at least 5 different concentrations), make 6 parallel wells for each concentration, and set up blank wells and control wells. During the drug action, use an inverted phase contrast microscope to observe the growth of the cells, in a 37-degree CO2 incubator After culturing in medium for 72 hours, add 20 μL of 5 g/L MTT to each well, incubate for 4 hours, discard the supernatant, add about 100 μL/well of DMSO, shake for 10 minutes to fully dissolve the purple crystals, and measure the absorbance at 490 nm with a microplate reader. The cytotoxicity of the test compound is expressed according to the value of IC50. The above values represent the average of 6 replicates.

All of the compounds 1-48 tested above were effective in inhibiting the growth of SK-Hep-1, HepG2, MKN-45, MDA-MB-231, A549, OVCAR-3, A498, HCT-116, M4 cells . Unexpectedly, most of them showed IC50 values less than 1 mM, some even less than 100 nM.

2. Drug sensitivity test

Before the experiment, pick Cryptococcus neoformans, Candida albicans, and Candida parapsilosis from the SDA medium stored at 4 degrees with an inoculation circle and inoculate it into 1mL YEPD culture solution, culture at 35 degrees, shake at 250r/min, and activate for 16 hours. Keep the fungus in late exponential growth phase. Take the bacteria solution into 1mL YEPD culture solution, and reactivate it with the above method. After 16 hours, count with a hemocytometer, and adjust the concentration of the bacteria solution to 1×10 ³ -5×10 ³ cells/mL with RPMI1640 culture solution.

Compounds 1-48 were respectively made into 6.4 mg/mL solutions in DMSO. Take a sterile 96-well plate, add 100 μL of RPMI1640 in each row of No. 1 wells as a blank control; add 100 μL of freshly prepared bacterial solution to each of No. 3-12 wells; add 200 μL of bacterial solution and 2 μL of test compound solution to No. 2 wells; Well No. 11 was diluted 10 times, and Well No. 12 contained no drug; it was used as a positive control. Each drug-sensitive plate was incubated at 35 degrees.

MIC value determination

After Candida and Cryptococcus neoformans were cultured at 35°C for 72 hours, the OD value of each well was measured at 630 nm with an enzyme label analyzer. Compared with the positive control wells, the drug concentration in the lowest concentration well whose OD value drops by more than 80% is MIC ₈₀ (drug concentration when 80% of fungal growth is inhibited).

Claims (7)

1. the compound shown in formula I or its pharmaceutically acceptable salt are for the preparation of antitumor and purposes antifungal drug, describedTumour is cancer of the stomach, liver cancer, oophoroma, kidney, colon cancer or melanoma, and described fungi is neogenesis cryptococcus or closely level and smoothCandida albicans, the structure of described formula I compound is as follows:

Wherein R₂、R₃、R₄For monosubstituted or polysubstituted, be selected from hydrogen, halogen, alkaneOxygen base, alkyl, hydroxyl, carboxyl, itrile group, refer to-O-of described alkoxyl alkyl, described alkyl is chosen as methyl, secondBase, n-propyl group, i-propyl group, n-butyl, i-butyl, t-butyl.

2. purposes according to claim 1, is characterized in that R₂、R₃、R₄Be selected from hydrogen, bromine, methoxyl group, chlorine, hydroxyl,Carboxyl, itrile group.

3. purposes according to claim 1, is characterized in that the compound that described formula I compound is following structure

4. purposes according to claim 1, is characterized in that R₂、R₃、R₄For monosubstituted.

5. purposes according to claim 1, is characterized in that described salt is inorganic salts or organic salt.

6. purposes according to claim 1, is characterized in that described salt is hydrochloride, sulfate, fumarate or sulfonic acidSalt.

7. purposes according to claim 1, is characterized in that described salt is mesylate.