A compound that inhibits BRM

Abstract

The present invention discloses a class of BRM ATPase inhibitor compounds as shown in formula (I), uses thereof and pharmaceutical compositions containing the same. The compounds have a strong inhibitory effect on BRM ATPase and have good application prospects in BRG1 gene mutation tumors.

Description

Technical Field

[0001] The present invention relates to a compound for inhibiting BRM, use thereof and a pharmaceutical composition containing the compound. Background Art

[0002] Cancer is one of the leading causes of death worldwide and is mainly caused by genetic and epigenetic changes. These changes affect the gene expression or signaling pathways of normal cells, leading to the transformation of normal cells into malignant tumor cells. Cancer genome sequencing results of tumor samples found that subunit genes of the human ATP-dependent chromatin remodeling complex SWI / SNF are mutated in >20% of human tumors, and most of them are loss-of-function mutations.

[0003] BRM (Protein brahma homolog) is the ATPase subunit of the SWI / SNF complex and is mutually exclusive with BRG1 (Protein brahma homolog 1), which is also an ATPase subunit. ATPase hydrolyzes ATP to provide energy for the SWI / SNF complex, enabling it to regulate gene expression and key cellular processes, and plays an important role in maintaining cell vitality.

[0004] The BRG1 gene (SMARCA4) is highly mutated in many types of cancer, including but not limited to melanoma, endometrial cancer, non-small cell lung cancer, esophageal and gastric adenocarcinoma, urothelial bladder cancer, and colorectal cancer. In contrast, the BRM gene (SMARCA2) rarely mutates. BRM and BRG1 are complementary and redundant in their functions related to cell viability, such as participating in the regulation of cell cycle and hormone action pathways, DNA damage repair response, etc. Therefore, in tumor cells with BRG1 gene defects, BRM can compensate for the functional loss of BRG1 and maintain the function of the SWI / SNF complex to support cell viability.

[0005] The synthetic lethality of BRM and BRG1 has been verified in in vitro and in vivo disease models of various types of tumor cells: degradation of BRM or silencing of BRM gene expression in BRG1 gene-deficient cells will inhibit tumor cell proliferation in vitro and tumor growth in vivo; BRG1 gene-deficient tumor cells are extremely sensitive to shRNA of the BRM gene; inhibiting the ATPase activity of BRM can inhibit the in vivo growth of BRG1 gene-deficient tumors... Therefore, in BRG1 gene mutant tumors, BRM is a potential drug treatment target.

[0006] Compared with BRG1, drug interference with BRM has a lower risk of toxicity. The ATPase activity and chromatin remodeling ability of the BRM-SWI / SNF complex are much lower than those of the BRG1-SWI / SNF complex. In normal cells, the SWI / SNF complex mainly relies on BRG1 to function. Homozygous knockout of the BRG1 gene does not cause embryonic lethality in mice or affect the reproductive capacity of adult mice, which is the opposite of the performance of homozygous knockout of the BRM gene.

[0007] Therefore, targeted inhibition of BRM ATPase activity has the potential to become a new treatment option for tumors with BRG1 gene mutations.

[0008] Currently, no BRM ATPase inhibitor drugs have entered the clinical stage, but some BRM ATPase inhibitors are in preclinical development. Foghorn’s recently disclosed patent WO2022103899 discloses a number of highly selective BRM ATPase inhibitors, the structural examples of which are as follows:

[0009] Summary of the invention

[0010] The present invention provides a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof:

[0011]

[0012] Among them, R1 is -CN, -OC 1-3 Alkyl, -NRR2R3, -SC 1-3 Alkyl, OH;

[0013] R2 and R3 are each independently C 1-3 alkyl.

[0014] In a preferred embodiment, R2 and R3 are each independently H or methyl.

[0015] In a more preferred embodiment, R1 is -CN, -OCH3, -NH2, -NHCH3, -N(CH3)2, -SCH3, or OH.

[0016] The compound or pharmaceutically acceptable salt thereof described in the present invention may be any of the following compounds:

[0017]

[0018] The present invention also provides the use of the compound shown above or a pharmaceutically acceptable salt thereof in the preparation of a BRM ATPase inhibitor, including the use in treating melanoma, endometrial cancer, non-small cell lung cancer, esophageal gastric adenocarcinoma, urothelial bladder cancer or colorectal cancer.

[0019] The present invention also provides a pharmaceutical composition, which comprises the compound shown above or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0020] As used herein, the term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt of a compound of the present invention. DETAILED DESCRIPTION

[0021] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0022] Example 1 EVO33354

[0023]

[0024] At room temperature, EVO33354-A1 (60 mg, 98.67 μmol, synthesized with reference to patent WO2022103899), zinc powder (12.9 mg, 197.33 μmol), zinc cyanide (23.17 mg, 197.33 μmol), Pd(dppf)Cl2.CH2Cl2 (16.12 mg, 19.73 μmol) and anhydrous 1,4-dioxane (5 mL) were added to the microwave tube and reacted at 100 ° C for 3 hours. After the reaction, water (50 mL) was added, ethyl acetate was extracted (3*20 mL), the organic phase was concentrated to dryness, and high pressure reverse phase was used to prepare yellow solid EVO33354 (20 mg, yield 33.86%), LCMS (ESI) m / z=599.33[M+H] + , purity: 99.1%. 1 H NMR(400MHz, DMSO-d6)δ9.78(t,J=5.8Hz,1H),9.41(s,1H),8.75(q,J=1.9Hz, 2H),8.65(q,J=8.7Hz,2H),7.94-7.85(m,2H),7.75(dd,J=8.5,7.4Hz,1H),7. 03(d,J=8.5Hz,1H),5.18(s,2H),4.84(d,J=5.8Hz,2H),4.35-4.24(m,4H),3. 86-3.79(m,2H),3.72-3.61(m,2H),2.55-2.44(m,2H),1.25(d,J=6.2Hz,6H).

[0025] Example 2 EVO33377

[0026]

[0027] Referring to Example EVO33354, the product was separated by HPLC preparative column and freeze-dried to obtain yellow solid EVO33377 (16 mg, yield: 53.72%), LCMS (ESI) m / z=604.47 [M+H] + , purity: 98.9%. 1 H NMR(400MHz,DMSO-d6)δ9.66(t,J=5.9Hz,1H),9.40(d,J=0.8Hz,1H),8.70-8.59(m,2H) ,8.17(t,J=1.5Hz,1H),7.96-7.88(m,2H),7.82(s,1H),7.74(dd,J=8.5,7.4Hz,1H),7.0 3(d,J=8.5Hz,1H),5.06(s,2H),4.83(d,J=5.8Hz,2H),4.35-4.28(m,2H),4.19(dd,J=6. 1,3.8Hz,2H),3.96(s,3H),3.75-3.61(m,4H),2.55-2.42(m,2H),1.21(d,J=6.2Hz,6H).

[0028] Example 3 EVO33419

[0029]

[0030] At room temperature, EVO33354-A1 (100 mg, 164.44 μmol), tert-butyl carbamate (96.32 mg, 822.22 μmol), cesium carbonate (160.74 mg, 493.33 μmol), XPhos (15.68 mg, 32.89 μmol), Pd2(dba)3 (15.06 mg, 16.44 μmol) and anhydrous 1,4-dioxane (4 mL) were added to the reaction bottle, argon gas was replaced (*3), and microwave was reacted at 110°C for 3 hours. After cooling to room temperature, the filtrate was concentrated under reduced pressure, purified by liquid phase preparation, and freeze-dried to obtain a light yellow solid EVO33419 (15 mg, yield: 15.5%), LCMS (ESI) m / z = 589.4 [M+H] + , purity: 99.0%. 1H NMR(400MHz,DMSO-d6)δ9.42(t,J=5.8Hz,1H),9.39(d,J=0.8Hz,1H),8.68-8.61(m,2H), 7.92(d,J=7.6Hz,1H),7.76-7.72(m,3H),7.53(d,J=1.6Hz,1H),7.03(d,J=8.4Hz,1H),5 .92(s,2H),4.93(s,2H),4.76(d,J=5.6Hz,2H),4.31(d,J=12.8Hz,2H),4.16(t,J=4.8Hz ,2H),3.73-3.62(m,2H),3.58(t,J=4.8Hz,2H),2.54-2.44(m,2H),1.21(d,J=6.4Hz,6H).

[0031] Example 4 EVO33420

[0032]

[0033] At room temperature, EVO33354-A1 (30 mg, 49.33 μmol), methylamine tetrahydrofuran solution (179.80 mg, 5.79 mmol, 0.2 mL), cesium carbonate (48.22 mg, 148.00 μmol), t-BuXphos (4.19 mg, 9.87 μmol), Pd2(dba)3 (4.52 mg, 4.93 μmol) and anhydrous tetrahydrofuran (2 mL) were added to the reaction bottle, replaced with argon (*3), and reacted at 70 ° C in a microwave for 2 hours. After cooling to room temperature, the mixture was filtered and the filtrate was concentrated to dryness under reduced pressure. The mixture was first purified by column chromatography (DCM:MeOH=20:1) and then purified by silica gel plate (DCM:MeOH=20:1) to obtain a light yellow solid EVO33420 (5.33 mg, yield: 17.9%). LCMS (ESI) m / z=603.4 [M+H] + , purity: 97.3%. 1H NMR (400MHz, DMSO-d6) δ9.51(t,J=5.8Hz,1H),9.40(d,J=0.8Hz,1H),8.68-8.61(m,2H),7.91(d,J=7.6Hz,1 H),7.80(d,J=2.0Hz,1H),7.78(s,1H),7.74(dd,J=8.4,7.2Hz,1H),7.41(d,J=1.6Hz,1H),7.03(d,J=8.8Hz ,1H),6.29-6.27(m,1H),4.97(s,2H),4.79(d,J=5.6Hz,2H),4.31(dd,J=12.8,2.4Hz,2H),4.18-4.16(m,2H ),3.70-3.64(m,2H),3.59(t,J=4.8Hz,2H),2.81(d,J=4.4Hz,3H),2.53-2.47(m,2H),1.21(d,J=6.0Hz,6H).

[0034] Example 5 EVO33422-B

[0035]

[0036] Referring to Example EVO33354, the product was separated by HPLC preparative column and freeze-dried to obtain a yellow solid EVO33422-B (2.38 mg, yield: 5.45%), LCMS (ESI) m / z=620.45 [M+H] + , purity: 90.1%. 1 H NMR(400MHz,DMSO-d6)δ9.69(t,J=5.9Hz,1H),9.40(d,J=0.8Hz,1H),8.70-8.59(m,2H),8.3 5(d,J=1.7Hz,1H),8.17(d,J=1.7Hz,1H),7.91(d,J=7.4Hz,1H),7.83(s,1H),7.75(dd,J=8. 5,7.4Hz,1H),7.03(d,J=8.5Hz,1H),5.16(s,2H),4.83(d,J=5.8Hz,2H),4.35-4.28(m,2H), 4.25-4.18(m,2H),3.75-3.63(m,4H),2.63(s,3H),2.54-2.45(m,2H),1.21(d,J=6.0Hz,6H).

[0037] Example 6 EVO33456

[0038]

[0039] At room temperature, EVO33354-A1 (50 mg, 82.22 μmol), water (0.1 mL), cesium carbonate (80.37 mg, 246.67 μmol), t-BuXphos (13.97 mg, 32.89 μmol), Pd2(dba)3 (15.06 mg, 16.44 μmol) and anhydrous tetrahydrofuran (4 mL) were added to the reaction bottle, argon gas was replaced (*3), and microwave heating was performed at 70 ° C for 1 hour. After cooling to room temperature, the mixture was filtered, washed with ethyl acetate, and the filtrate was concentrated to dryness under reduced pressure. Liquid phase preparative purification was performed to obtain a light yellow solid EVO33456 (20.54 mg, yield: 42.4%), LCMS (ESI) m / z = 590.20 [M+H] + , purity: 99.17%. 1 H NMR(400MHz,DMSO-d6)δ9.53(t,J=6.0Hz,1H),9.39(s,1H),8.67-8.60(m,2H),7.95(d ,J=1.6Hz,1H),7.91(d,J=7.2Hz,1H),7.77-7.70(m,3H),7.03(d,J=8.8Hz,1H),5.03( s,2H),4.77(d,J=5.6Hz,2H),4.31(dd,J=13.2,2.4Hz,2H),4.17(dd,J=6.2,3.6Hz,2H ),3.70-3.65(m,2H),3.62(t,J=4.8Hz,2H),2.53-2.47(m,2H),1.21(d,J=6.0Hz,6H).

[0040] Effect Example 1 Inhibitory Activity Detection Experiment of Compounds on BRM and BRG1 ATPase

[0041] To characterize the ATPase inhibitory activity of compounds against BRM and BRG1, ADP-Glo TMKinase Assay (Promega, #V9101) detects the amount of ADP in the ATPase reaction endpoint system to evaluate ATPase activity in two steps: First, after the ATPase reaction, ADP-Glo ​​Reagent is added to terminate the reaction and deplete the remaining ATP; then, Kinase Detection Reagent is added to convert ADP into ATP, and ATP is consumed through the luciferase / luciferin reaction to produce a chemiluminescent signal, which is proportional to the concentration of ATP (converted from ADP generated by the ATPase reaction). Compared with the control wells without compound treatment, the ATPase inhibitory activity of the compound is evaluated by the reduction of the luminescent signal. The IC is calculated based on the data relationship between the compound concentration and the percentage of ATPase activity. 50 value.

[0042] The specific operation is as follows: the test compound (dissolved in DMSO, concentration 10uM-1nM, 50nL / well) is added to a 384-well plate (Greiner#781280). 2.5μL of a mixed solution of BRM truncations (expressed and purified BRM ATPase-SnACDomain, refer to the literature description DOI:10.1021 / acs.jmedchem.8b01318) or BRG1 truncations (expressed and purified BRG1ATPase-SnAC Domain, refer to the literature description DOI:10.1021 / acs.jmedchem.8b01318) and DNA (Invitrogen#15632011) is added to each well (protein final concentration 30nM, DNA final concentration 80nM). 2.5μL of ATP (Promega#V9101, final concentration 400uM) is added to each well to start the enzyme reaction. The system buffer is 20mM Tris, pH 8.0, 50mM NaCl, 20mM MgCl2, 0.1% Tween-20, 1mM DTT, the total volume is 5μL, and the reaction is carried out at 25°C for 60 minutes. Then, ADP Glo reagent (5μL) is added and incubated at 25°C for 90 minutes. Then, Kinase Detection Reagent (10μL) is added and incubated at 25°C for 60 minutes. Finally, chemiluminescence is detected.

[0043] The results of enzyme activity determination are listed in the following table:

[0044]

[0045] Where A represents IC 50 ≤100nM; B means 100nM<IC 50 ≤500nM; C means 500nM<IC 50≤1000nM; D means 1000nM<IC 50 ≤5000nM; E means 5000nM<IC 50 ≤10000nM; F represents IC 50 >10000nM; NA means not detected.

Claims

1. A compound as described below or a pharmaceutically acceptable salt thereof:

2. A pharmaceutical composition, characterized in that It comprises the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

3. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a BRM ATPase inhibitor.

4. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating melanoma, endometrial cancer, non-small cell lung cancer, esophageal gastric adenocarcinoma, urothelial bladder cancer or colorectal cancer.

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