CN108653282B - Application of benzothiazole and benzopyrrole compounds in preparation of antitumor drugs - Google Patents

Abstract

The invention provides benzothiazole compounds and benzopyrrole compounds, a pharmaceutical composition taking the benzothiazole compounds and the benzopyrrole compounds as active ingredients, and application of the benzothiazole compounds and the benzopyrrole compounds as Wee1 kinase inhibitors and in preparing anti-cancer therapeutic drugs. Wee1 has been shown to enhance its anti-tumor activity when used in conjunction with DNA damaging agents. The invention provides application of benzothiazole compounds and benzopyrrole compounds in preparation of Wee1 inhibitors. The series of compounds are proved to have obvious anti-tumor cell proliferation activity through in vitro anti-tumor cell activity screening experiments.

Description

Application of benzothiazole and benzopyrrole compounds in preparation of antitumor drugs

Technical Field

The invention relates to the technical field of medicines, in particular to benzothiazole compounds and benzopyrrole compounds, a pharmaceutical composition taking the benzothiazole compounds and the benzopyrrole compounds as active ingredients, application of the benzothiazole compounds and the benzopyrrole compounds in preparing Wee1 kinase inhibitors and application of the benzothiazole compounds and the benzopyrrole compounds in preparing antitumor drugs.

Background

Wee1 protein kinase was first found in fission yeast cells, belongs to the serine, threonine protein kinase family, is highly conserved and widely present in eukaryotes. It is a cell cycle regulatory protein that is activated primarily in the S phase or G2/M phase of the cell cycle. The novel polypeptide can regulate and control the phosphorylation state of cyclin dependent kinase 1(CDK1), so that the activity of a CDK1 and cyclin B (cyclin B) complex is regulated, the regulation and control of a cell cycle are realized, and the novel polypeptide has an important regulation effect on a DNA damage check point. Wee1 protein kinases regulate mitosis through the action of 3 cellular checkpoints, the cellular G2/M phase checkpoint, the cell size checkpoint and the cellular DNA damage checkpoint. Research has shown that over-expression of Wee1 kinase is detected in many types of malignant cells, indicating that Wee1 kinase is a candidate target for cancer therapy.

Currently, there are three major inhibitors based on Wee1 kinase, MK-1775, PD0407824, PD0166285, respectively. MK-1775 in combination with other chemotherapeutic drugs for ovarian cancer is currently in phase II clinical studies, whereas PD0166285 and PD0407824 inhibitors have not been tested in patients. The Wee1 inhibitor exerts an anti-tumor effect by inhibiting the activity of Wee1, which leads tumor cells to enter M phase from S phase or G2 phase in advance, cannot complete DNA synthesis and repair, and is characterized by abnormal histone synthesis of cells, disfunction of formation of a dipole spindle, chromosome dispersion, and finally exit from mitosis, causing apoptosis, which is called "mitotic catastrophe". Wee1 inhibitor has been applied to clinical stage, and has initially demonstrated wide application prospect in antitumor therapy, and the combined use of Wee1 kinase inhibitor and DNA inhibitory drug is probably a more effective therapeutic strategy against cancer. Therefore, the discovery of the Wee1 small-molecule inhibitor with higher efficiency and specificity has great practical significance in tumor treatment.

At present, no reports related to benzothiazole compounds and benzopyrrole compounds and pharmaceutical compositions thereof in preparation of Wee1 kinase inhibitors and antitumor drugs are found in the prior art.

Disclosure of Invention

The invention aims to provide benzothiazole compounds and benzopyrrole compounds I and II, a pharmaceutical composition taking the benzothiazole compounds and the benzopyrrole compounds as active ingredients, a preparation method of the benzothiazole compounds and the benzopyrrole compounds, and application of the benzothiazole compounds and the benzopyrrole compounds in preparation of Wee1 kinase inhibitors and antitumor drugs.

In order to achieve the above purpose of the present invention, the present invention provides the following technical solutions:

a pharmaceutical composition comprises a therapeutic amount of benzothiazole and benzopyrrole derivatives represented by the following structural formulas (I) and (II) or pharmaceutically acceptable salts thereof, and at least one pharmaceutically acceptable carrier

The pharmaceutical composition of the present invention, wherein the pharmaceutically acceptable salts of benzothiazole and benzopyrrole compounds are salts of compounds with acids or sodium, potassium, magnesium and calcium salts of compounds with acidic protons replaced by metal ions; the salt formed by the acid is hydrochloride, hydrobromide, methanesulfonate, sulfate, phosphate, acetate, trifluoroacetate, trifluoromethanesulfonate, p-toluenesulfonate, tartrate, maleate, fumarate, succinate or malate.

The application of the pharmaceutical composition in preparing a Wee1 kinase inhibitor.

The application of the pharmaceutical composition in preparing anti-tumor drugs is characterized in that the tumors are head and neck tumors, respiratory system tumors, digestive system tumors, urinary system tumors, skeletal system tumors, gynecological tumors, hematological system tumors and other types of tumors.

The use of the pharmaceutical composition for the preparation of an anti-tumor medicament, wherein the tumor of the head and neck is thyroid cancer, nasopharyngeal cancer, meningeal cancer, acoustic neuroma, pituitary tumor, oral cancer, craniopharyngioma, thalamus and brain stem tumor, angiogenetic tumor or intracranial metastatic tumor; the respiratory system tumor is lung cancer; the digestive system tumor is liver cancer, gastric cancer, esophageal cancer, colorectal cancer, rectal cancer, colon cancer or pancreatic cancer; the urinary system tumor is renal cancer, bladder cancer, prostatic cancer or testicular cancer; the skeletal system tumor is bone cancer; the gynecological tumor is breast cancer, cervical cancer or ovarian cancer; the blood system tumor is leukemia, malignant lymphoma or multiple myeloma.

The application of the pharmaceutical composition in preparing antitumor drugs. Wherein said other type of tumor is malignant melanoma, glioma or skin cancer.

The pharmaceutical composition as described, characterized in that it is in the form of a preparation suitable for oral or injectable administration.

The pharmaceutical composition of claim, wherein the orally administered formulation is in the form of tablets, sustained release tablets, controlled release tablets, lozenges, hard or soft capsules, dripping pills, pellets, aqueous or oily suspensions, emulsions, dispersible powders or granules, oral solutions, syrups or elixirs, and the parenterally administered formulation is in the form of sterilized aqueous or oily solutions, sterile powders, liposomes, emulsions, microemulsions, nanoemulsions or microcapsules.

The invention also provides application of the pharmaceutical composition in preparation of a Wee1 kinase inhibitor.

And the application of the pharmaceutical composition in preparing antitumor drugs. The use as described, wherein the tumor is leukemia, liver cancer, lung cancer, breast cancer, colon cancer.

The compound of the invention is tested by antitumor activity tests to obtain the IC of five tumor cell strains of leukemia, liver cancer, lung cancer, breast cancer and colon cancer₅₀. The results show that the compounds I and II have certain in vitro tumor growth inhibition activity.

When used as a medicament, the compounds of the present invention may be used as such or in the form of a pharmaceutical composition. The pharmaceutical composition contains 0.1-99%, preferably 0.5-95% of the compound of the invention, and the balance of pharmaceutically acceptable salts, or pharmaceutically acceptable carriers and/or excipients which are nontoxic to human and animals.

The administration amount of the compound of the present invention may vary depending on the route of administration, age, body weight of the patient, type and severity of the disease to be treated, etc., and the daily dose thereof may be 0.01 to 10mg/kg body weight, preferably 0.1 to 5mg/kg body weight. One or more administrations may be carried out.

It can be administered orally in the form of solid or liquid, such as powder, tablet, coated preparation, capsule, solution, syrup, or dripping pill.

The injection can be solid or liquid preparation, such as powder injection, solution injection, etc.

Drawings

FIG. 1 is a benzothiophene of the present inventionIC of azole compound I on lung cancer A546 cell strain₅₀In the form of a graph of the graph,

FIG. 2 shows IC of Compound II of the present invention on human leukemia Jurkat cell line₅₀Graph is shown.

Detailed Description

The following embodiments of the present invention will be described in detail with reference to the accompanying drawings, but the present invention is not limited thereto.

Example 1:

the compounds of the invention are partially purchased from IBScreen, and compounds I and II are numbered correspondingly in the library: STOCK 1S-28804; STOCK 6S-58881.

The structural formulae of compounds I and II are shown below:

example 2:

half-growth inhibition concentration IC of compound of the invention on five different tumor cell strains₅₀The determination of (1):

1. principle and procedure of experiment

MTS is MTT analogue, succinate dehydrogenase in mitochondria of living cells can metabolize and reduce MTS to generate soluble Formazan (Formazan) compound, and the soluble Formazan compound can be directly dissolved in a culture medium. When in detection, only a small amount of CellTiter is needed

The Aqueous One Solution reagent is directly added into the culture medium in the culture plate hole, incubated for 1-4h, and then the absorbance value of 490nm is read by a microplate reader. The optical density OD (490nm) of the compound is proportional to the number of living cells. Cisplatin and paclitaxel were used as positive controls for the assay. IC of the Compound₅₀Values were determined by concentration effect growth curve calculation.

Standing at room temperature for 90 minutes to completely melt CellTiter

Aqueous One SolutAnd ion agent. Five different tumor cell lines with optimal cell concentrations were selected, including Jurkat, SMMC-7721, A-549, MCF-7 and SW 480. Spreading into 96-well culture plate with each well at 37 deg.C and 100 μ L, 5% CO₂Incubated for 24h under the conditions of (1), and 20 mu of LCellTiter was added

After incubation for 2h, absorbance values were read at 490 nm. And setting positive control groups of cisplatin and paclitaxel.

2. Reagent and instrument

Reagent: CellTiter

AQueous One Solution Cell Proliferation Assay instrument: CO 2₂Incubator (Thermo), clean bench (Thermo), chemiluminescence enzyme-labeling instrument (Thermo)

3. Data processing

The inhibition rate of the compound on monoamine oxidase B was calculated in Excel according to the following formula (1). Formula (1):

inhibition rate: inhibition rate (1-a sample/a positive control) × 100%

The IC of the compound for inhibiting tumor cell strains is calculated in GraphPad Prism 5 according to the following formula (2)₅₀The value is obtained.

Formula (2):

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC₅₀-X)*Hill Slope))

y is the inhibition rate and X is the compound concentration.

4. Results of the experiment

IC of Compounds I and II on different tumor cell lines₅₀As shown in table 1.

TABLE 1 IC of Compounds on different tumor cell lines₅₀(μM)

Example 3:

adding 4% ethanol sulfate solution to compound I or II, filtering, drying, and making into sulfate compound.

Example 4:

compound I or II was added to a 4% hydrochloric acid solution at pH 4, filtered, and dried to obtain the hydrochloride compound.

Example 5:

compound I or II was added to a 4% tartaric acid solution at pH 4, filtered, and dried to give a tartrate compound.

Example 6:

compound I or II was added with 4% citric acid solution at pH 4, filtered and dried to obtain a citrate compound.

Example 7:

and (3) capsule preparation: 10mg of compound I or II or the salt obtained in example 3-6, 187mg of lactose, 3mg of magnesium stearate.

The preparation method comprises the following steps: mixing compound I or II or its salt with cosolvent, sieving, mixing, and encapsulating the obtained mixture into hard gelatin capsules with each capsule weight of 200mg and active ingredient content of 10 mg.

Example 8:

and (3) tablet preparation: adding 10mg of compound I or II or salt prepared in examples 3-6 into 180mg of lactose, 55mg of starch and other auxiliary materials, uniformly mixing, wetting with water, granulating, drying, adding 5mg of magnesium stearate, uniformly mixing, and tabletting, wherein each tablet is about 250 mg.

Example 9:

an ampoule agent: 2mg of Compound I or II or the salt obtained in example 3 to 6 and 10mg of sodium chloride were dissolved in an appropriate amount of water for injection, and the resulting solution was filtered and aseptically filled in an ampoule.

Example 10:

freeze-dried preparation for injection: 10mg of compound I or II or the salt obtained in example 3-6, 2mg of sodium hydrogencarbonate and 252mg of mannitol were added.

The preparation method comprises the following steps: dissolving sodium bicarbonate and mannitol in water for injection, adsorbing with activated carbon for 30min to remove pyrogen, filtering to remove activated carbon, adding compound or its salt into the filtrate, ultrasonic treating to dissolve, adjusting pH to 5.0-7.0 with 1N hydrochloric acid, filtering with microporous membrane, adding water for injection, packaging, freeze drying, plugging, and capping.

Example 11:

and (3) capsule preparation: 10mg of compound I or II or the salt prepared in example 3-6, 187mg of lactose and 3mg of magnesium stearate are added.

The preparation method comprises the following steps: mixing compound I or II or its salt with cosolvent, sieving, mixing, and encapsulating the obtained mixture into hard gelatin capsules with each capsule weight of 200mg and active ingredient content of 10 mg.

Claims (4)

1. The application of the pharmaceutical composition in preparing antitumor drugs is characterized in that the tumors are leukemia, liver cancer, lung cancer, breast cancer and colon cancer, and the pharmaceutical composition contains therapeutic amount of benzothiazole and benzopyrrole derivatives shown in structural formulas (I) and (II) or pharmaceutically acceptable salts thereof and at least one pharmaceutically acceptable carrier,

2. the use of the pharmaceutical composition according to claim 1 for the preparation of an antitumor medicament, wherein the pharmaceutically acceptable salts of the benzothiazole and benzopyrrole compounds are salts of the compounds with acids; the salt formed by the acid is hydrochloride, hydrobromide, methanesulfonate, sulfate, phosphate, acetate, trifluoroacetate, trifluoromethanesulfonate, p-toluenesulfonate, tartrate, maleate, fumarate, succinate or malate.

3. The use of a pharmaceutical composition according to claim 1 for the preparation of an antitumor medicament, characterized in that the formulation is in the form of an oral or injectable preparation.

4. The use of the pharmaceutical composition according to claim 3 for the preparation of an antitumor agent, wherein the formulation for oral administration is in the form of tablets, troches, hard or soft capsules, dripping pills, pellets, aqueous or oily suspensions, emulsions, dispersible powders or granules, oral solutions, syrups or elixirs, and the formulation for injection is in the form of sterilized aqueous or oily solutions, sterile powders, liposomes, emulsions or microcapsules.

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