CN111018832A

CN111018832A - Preparation and application of imidazolone compounds containing tetrahydroisoquinoline structure

Info

Publication number: CN111018832A
Application number: CN201911170928.3A
Authority: CN
Inventors: 张华�; 刘凯璐; 闫薛; 朱孔凯
Original assignee: University of Jinan
Current assignee: University of Jinan
Priority date: 2019-11-26
Filing date: 2019-11-26
Publication date: 2020-04-17

Abstract

The invention discloses imidazolone compounds containing a tetrahydroisoquinoline structure and an application of the imidazolone compounds as a novel protein arginine methyltransferase 5 inhibitor, wherein the imidazolone compounds are shown in a figure 1.

Description

Preparation and application of imidazolone compounds containing tetrahydroisoquinoline structure

Technical Field

The invention relates to the fields of pharmaceutical chemistry and pharmacotherapeutics, in particular to preparation and application of an imidazolone compound containing a tetrahydroisoquinoline structure.

Background

Protein arginine methyltransferases (PRMTs) -catalyzed methylation of protein arginine is an important post-translational modification that uses S-adenosylmethionine (SAM) as a methyl donor to methylate and modify the nitrogen atom of the protein arginine side chain. The substrates of PRMTs are glycine and arginine rich proteins. The 9 human PRMTs identified (PRMT 1-9) were further subdivided into type I, type II and type III, depending on the status of the methylated arginine. Type I PRMT catalyzes the formation of monomethyl arginine (MMA) and asymmetric dimethyl arginine (aDMA), including PRMT1, PRMT2, PRMT3, PRMT4, PRMT6 and PRTM 8; type ii PRMT catalyzes the formation of MMA and symmetric dimethylarginine (sDMA), including PRMT5 and PRMT 9; type iii PRMT catalyzes the formation of MMA, including PRMT 7. PRMT5 belongs to type ii PRMT, transferring a methyl group from the SAM to the guanidine nitrogen of an arginine residue to produce MMA and sDMA. Human tissue proteomics analysis showed that PRMT1, -4 and-5 expression was more prevalent compared to PRMT2, -3, -6, -7 and-8. PRMT5 mediates various cellular processes such as transcription, RNA metabolism, signal transduction, and cell differentiation through symmetric dimethylation of multiple substrate proteins in the nucleus and cytoplasm.

Interestingly, increasing research has shown that PRMT5 is an attractive target for cancer therapy, which is overexpressed or upregulated in a variety of human cancers, including leukemia, lymphoma, breast cancer, lung cancer, bladder cancer, gastric cancer, pancreatic cancer, prostate cancer, colon cancer, multiple myeloma, liver cancer, melanoma, head and neck cancer, thyroid cancer, renal cell carcinoma, glioblastoma, or testicular cancer. Selective inhibition of PRMT5 was reported to block B cell transformation. In addition, PRMT5 controls melanoma growth through the SKI/SOX10 regulatory axis. PRMT5 is considered to be an effective target for the treatment of Mantle Cell Lymphoma (MCL), glioblastoma, multiple myeloma, and mixed leukemia (MLL) -acute myeloid leukemia.

LLY-283, Sinefungin and A9145C are reported to be SAM binding site inhibitors, EPZ015666 and its analogue GSK3326595 are substrate binding site inhibitors, EPZ015666 is the first PRMT5 inhibitor with potent activity in both in vitro and in vivo models of mantle cell lymphoma. In addition, clinical trials for GSK3326595 in phase i for the treatment of solid and non-hodgkin's lymphoma are ongoing. Although there are more than 10 PRMT5 inhibitors reported, most exhibit low potency or lack in vivo activity.

In view of the above, there is an urgent need in the art to develop more and more effective novel PRMT5 inhibitors.

Disclosure of Invention

In order to solve the problems, the invention provides an imidazolone compound containing a tetrahydroisoquinoline structure, a preparation method thereof and application thereof in preparing a medicament for preventing and/or treating cancer-related diseases.

The invention is realized by the following technical scheme.

In a first aspect of the invention, imidazolone compounds with novel structures are provided, and the general formula is shown in figure 1, wherein R is methyl, methoxy, trifluoromethyl and chlorine.

Among them, a preferred structure is shown in fig. 2.

The preparation method of the compound related to the invention has a preparation route shown in figure 4.

Preferably, the cancer to be prevented and/or treated is leukemia, lymphoma, breast cancer, lung cancer, bladder cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, multiple myeloma AML, liver cancer, melanoma, head and neck cancer, thyroid cancer, renal cell carcinoma, glioblastoma or testicular cancer.

Preferably, the related disease is a disease associated with an abnormal arginine methyltransferase 5 activity.

Has the advantages that: the compound has inhibitory activity to PRMT5, the preparation method of the compound is simple and easy to implement, the compound has tumor cell proliferation inhibitory activity, and the raw materials are easy to obtain.

Drawings

FIG. 1 is a new class of imidazolone compounds.

FIG. 2 shows preferred novel imidazolones.

FIG. 3 shows the structural design of compounds Ac-1 to Ac-11.

FIG. 4 shows a preparation route of compounds Ac-1 to Ac-11.

FIG. 5 shows the enzyme inhibitory activity of compounds Ac-1-Ac-11 tested by the radioisotope method.

FIG. 6 Effect of compounds Ac-1-Ac-11 on Z-138 cell proliferation.

Detailed Description

The following examples are given for the detailed implementation and specific operation of the present invention, but the scope of the present invention is not limited to the following examples.

Example 1: and structural design of the compounds Ac-1-Ac-11.

The compounds Ac-1-Ac-11 are obtained by utilizing a traditional structure optimization mode, and the structure design is shown in figure 3. Wherein R is methyl, methoxy, trifluoromethyl and chlorine.

Example 2: and preparing compounds Ac-1-Ac-11.

The compounds Ac-1-Ac-11 are synthesized by substituted phenyl isonitrile, and the substituted phenyl isonitrile reacts with aminoacetaldehyde dimethyl acetal at room temperature to obtain 1- (2, 2-dimethoxyethyl) -3-phenylurea; the crude 1- (2, 2-dimethoxyethyl) -3-phenylurea was dissolved in 5ml acetonitrile, 2ml water and 2ml trifluoroacetic acid, the mixture was stirred at room temperature for 6 hours, the solvent was removed under reduced pressure, and the mixture was purified by chromatography on silica gel column (petroleum ether/ethyl acetate 1/1) to give 1-phenyl-1, 3-dihydro-2H-imidazol-2-one; dissolving 1-phenyl-1, 3-dihydro-2H-imidazole-2-ketone in N, N-dimethylformamide, adding sodium hydride in ice bath, stirring for half an hour, adding epoxy chloropropane, stirring at room temperature for 6 hours, extracting the reaction mixture with ethyl acetate after the reaction is finished, washing with water, concentrating an organic layer, and performing chromatographic purification by using a silica gel column (petroleum ether/acetone 3/2) to obtain a corresponding epoxy compound; dissolving an epoxy compound in ethanol, adding 1,2,3, 4-tetrahydroisoquinoline, reacting for 15 minutes, removing the solvent under reduced pressure, and performing chromatographic purification by using a silica gel column (petroleum ether/acetone 2/1) to obtain compounds Ac-1-Ac-11. The specific preparation route is shown in figure 4, wherein (a): stirring aminoacetaldehyde dimethyl acetal at room temperature; (b) the method comprises the following steps Acetonitrile, trifluoroacetic acid and water are stirred at room temperature; (c) the method comprises the following steps N, N-dimethylformamide, ice bath and sodium hydride.

Example 3: inhibitory activity of compounds Ac-1-Ac-11 on PRMT 5.

The compounds were tested for enzyme inhibitory activity by means of radioisotopes. The specific experimental steps are as follows:

(1) preparing 1x experiment buffer (modified Tris-HCl buffer);

(2) diluting the compound to the desired concentration in a 96-well plate;

(3) preparing a protein solution, and using 1x experiment buffer solution;

(4) adding a substrate into 1x experiment buffer solution to prepare a substrate solution;

(5) will 2³H]Preparation of [ 2 ], [ SAM ] by adding to 1X test buffer³H]-a SAM solution;

(6) adding SAM to 1x experiment buffer solution to prepare cold SAM solution;

(7) removing 10. mu.L of the protein solution into a 96-well plate containing the compound;

(8) incubation for 15 minutes at room temperature;

(9) add 10. mu.L of substrate solution to each well;

(10) to each well was added 10. mu.L of³H]-the SAM solution initiates the reaction;

(11) incubation at room temperature for 240 min;

(12) add 10. mu.L of cold SAM solution to each well to stop the reaction;

(13) transferring 40 mu L of reaction mixed solution to a GF/B plate, and washing for 3 times by using triple distilled water in vacuum;

(14) reading data on a Microbeta liquid scintillation/luminescence counter;

(15) the inhibition rate was calculated according to the formula% Inh = (maximum signal-compound signal)/(maximum signal-minimum signal) × 100, the maximum signal being obtained from the reaction of the enzyme and the substrate and the minimum signal being obtained from the substrate. Data were plotted after processing using GraphPadPrism 5.0. EPZ015666 was used as a positive control. The results of the experiment are shown in FIG. 5.

Example 4: effect of Compounds Ac-1 to Ac-11 on cell proliferation.

Cell culture: the culture medium for Z-138 cell culture is RPMI 1640 + 10% fetal bovine serum, and 100U/mL penicillin and 100. mu.g/mL streptomycin are added to the culture medium to prevent bacterial contamination, and 5% CO is added at 37 deg.C₂The cells are cultured under the saturated humidity condition, and the cells for experiments are all in the exponential growth phase.

And (3) detecting cell proliferation activity: adjusting the cell concentration to 1X 10⁵Inoculating to 24-well culture plate with 1 mL of each well volume, setting control group and experimental group, adding DMSO into the control group, and adding PRMT5 small molecule into the experimental groupInhibitor is added to make the final concentration reach 0-200 mu M, and the mixture is placed at 37 ℃ and 5% CO₂The cells were cultured in an incubator, the amount of viable cells was measured every four days using CellTiter-Glo kit, and data was recorded on day 8. The results of the experiment are shown in FIG. 6.

Claims

1. A kind of imidazolone compound containing tetrahydroisoquinoline structure is shown in figure 1, wherein R is methyl, methoxy, trifluoromethyl and chlorine.

2. A process for the preparation of a compound according to claim 1, wherein the desired product is prepared by the route of figure 4.

3. Use of a compound according to claim 1 for the preparation of a medicament for the prophylaxis and/or treatment of cancer or a related disease.

4. The use according to claim 3, wherein the cancer to be prevented and/or treated is leukemia, lymphoma, breast cancer, lung cancer, bladder cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, multiple myeloma AML, liver cancer, melanoma, head and neck cancer, thyroid cancer, renal cell carcinoma, glioblastoma or testicular cancer.

5. The use according to claim 3, wherein the related disease is a disease caused by abnormal activity of protein arginine methyltransferase 5.