A quinazoline amine compound, its preparation method and application

Abstract

The application discloses a quinazoline amine compound, a preparation method and application thereof. Specifically, the application provides a compound with a general formula I and a pharmaceutically acceptable salt thereof, which has good inhibiting effect on SOS1 and EGFR and can be used for treating various cancers.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, specifically relating to a quinazolinamine compound, its preparation method, and its application. Background Technology

[0002] SOS1 (Son of sevenless homologue 1) protein is a guanine nucleotide exchange factor (GEF) widely expressed in humans and associated with various human cancers and other diseases. In human cells, activated SOS1 promotes the conversion between inactive RAS-GDP and active RAS-GTP, while activated RAS (Rat Sarcoma Virus) can activate multiple downstream signaling pathways, such as RAF-MEK-ERK, PI3K-AKT-mTOR, and Ral-GDS, regulating cell survival and proliferation. Inhibiting SOS1 protein can suppress RAS protein activation, thereby inhibiting tumor development and progression. Research on SOS1 inhibitors primarily focuses on developing ortho-SOS1 inhibitors. Currently, SOS1 inhibitors are still in the early stages of development, with only MRTX0902 and BI1701963 having entered clinical trials.

[0003] Epidermal growth factor receptor (EGFR) is a member of the human epidermal growth factor receptor (HER) family. EGFR is closely related to tumor proliferation, invasion, and metastasis. EGFR dimers upon binding to epidermal growth factor (EGF) or transforming growth factor-α (TGFα). Activated EGFR stimulates downstream pathways, including the RAS-RAF-MEK-ERK, PI3K-AKT-mTOR, and JAK-STAT pathways, leading to tumor growth. Therefore, targeting EGFR is an effective treatment for tumors. Currently, EGFR inhibitors such as gefitinib have been approved for marketing.

[0004] EGFR and SOS1 are two proteins located on the same signaling pathway. Studies have shown that combined inhibition of SOS1 and EGFR has a stronger anti-tumor effect against KRAS-mutant tumors. For complex diseases involving abnormalities in multiple signaling pathways, combination therapy has better anti-tumor efficacy than single-target combination therapy. However, combination therapy has problems such as toxicity, poor compliance, and drug-drug interactions. Therefore, the development of novel small molecule inhibitors targeting both SOS1 and EGFR with good anti-tumor activity is of great significance. Summary of the Invention

[0005] The purpose of this invention is to provide a dual-target compound of SOS1 and EGFR to address the problem of poor efficacy of current SOS1 inhibitors and EGFR inhibitors in tumor treatment.

[0006] The technical solution of the present invention is as follows:

[0007] A quinazolinamine compound as shown in Formula I and its pharmaceutically acceptable salt have the following structure:

[0008]

[0009] Ar selected Among them, R 1 –R 3 Each is independently selected from hydrogen, deuterium, halogen, methyl, trifluoromethyl, cyano, and amino.

[0010] Preferably, the quinazolinamine class of Formula I and its pharmaceutically acceptable salt are any of the following compounds:

[0011]

[0012]

[0013] A pharmaceutical composition comprising the compound of claims 1 to 2, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or thereof.

[0014] The use of the quinazolinamine compounds and their pharmaceutically acceptable salts, as well as the pharmaceutical compositions described herein, in the preparation of SOS1 / EGFR dual-target inhibitor drugs.

[0015] The use of the quinazolinamine compounds and their pharmaceutically acceptable salts as described in claims 1-2, and the pharmaceutical compositions thereof, in the preparation of medicaments for the prevention and / or treatment of tumors.

[0016] Beneficial effects:

[0017] Compared with the prior art, the present invention has the following significant features: The present invention discloses a new compound represented by general formula I, which can effectively inhibit the targets of SOS1 and EGFR and can be used to prepare drugs for treating cancer or tumor-related diseases; the present invention also discloses a method for preparing the compound of general formula I. Detailed Implementation

[0018] To better understand the present invention, the following embodiments are provided to further illustrate the invention, but the scope of the invention is not limited to the following embodiments.

[0019] Example 1

[0020]

[0021] Synthesis route:

[0022]

[0023] Synthesis of compound I-1

[0024] I-1a (100.0 mg, 0.42 mmol), I-1b (141.5 mg, 0.42 mmol, 1.0 eq), and N,N-diisopropylethylamine (162.9 mg, 1.26 mmol, 3.0 eq) were dissolved in 1.0 mL of DMSO. The reaction mixture was stirred overnight at 60 °C, diluted with water, and extracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate. The crude product was purified by column chromatography to give a pale yellow solid I-1 (131.5 mg, 65% yield). 1 H NMR(300MHz, DMSO-d6)δ8.31(s,1H),8.25(d,J=7.8Hz,1H),7.78(s,1H),7.20(s,1H),6.87–6.80(m,2H),6.72–6.67(m,1H),5.65–5 .44(m,3H),4.16(t,J=5.9Hz,2H),3.89(s,3H),3.70–3.55(m,4H),2.71–2.41(m,8H),2.02(t,J=6.1Hz,2H),1.56(d,J=7.1Hz,3H).

[0025] Example 2

[0026]

[0027] Following the synthetic method of compound I-1, replacing (R)-3-(1-aminoethyl)-5-(trifluoromethyl)aniline with (R)-1-[3-(trifluoromethyl)phenyl]ethylamine yielded compound I-2 (42.6 mg, 40% yield). ¹H NMR (300 MHz, CDCl₃) δ 8.51 (s, ¹H), 7.73–7.68 (m, ¹H), 7.68–7.62 (m, ¹H), 7.57–7.50 (m, ¹H), 7.46 (dd, J = 7.6, 7.5 Hz, ¹H), 7.19 (s, ¹H), 7.03 (s, ¹H), 5.81 (d, J = 7.2 Hz). ,1H),5.76–5.64(m,1H),4.17(t,J=6.6Hz,2H),3.96(s,3H),3.78–3.70(m,4H),2 .59(t,J=7.1Hz,2H),2.55–2.45(m,4H),2.17–2.04(m,2H),1.72(d,J=6.8Hz,3H).

[0028] Example 3

[0029]

[0030] Following the synthetic method of compound I-1, (R)-3-(1-aminoethyl)-5-(trifluoromethyl)aniline was replaced with (R)-3-methyl-phenylethylamine to obtain compound I-3 (35.8 mg, yield 65%). 1 H NMR (300MHz, CDCl3) δ8.55(s,1H),7.30–7.25(m,4H),7.19(s,1H),7.15–7.09(m,1H),6.94(s,1H),5.72–5.57(m,2H),4.15(t,J=6.6Hz ,2H),3.97(s,3H),3.78–3.68(m,4H),2.57(t,J=7.1Hz,2H),2.53–2.45(m,4H),2.36(s,3H),2.16–1.99(m,2H),1.69(d,J=6.5Hz,3H).

[0031] Example 4

[0032]

[0033] By replacing (R)-3-(1-aminoethyl)-5-(trifluoromethyl)aniline with R-methylbenzylamine, compound I-4 (60.7 mg, 70% yield) can be obtained. 1H NMR(300MHz, CDCl3)δ8.54(s,1H),7.50–7.43(m,2H),7.41–7.33(m,2H),7.33–7.28(m,1H),7.18(s,1H),6.96(s,1H),5.79–5.61(m,2H), 4.13(t,J=6.6Hz,2H),3.96(s,3H),3.77–3.67(m,4H),2.55(t,J=7.1Hz,2H),2.51–2.43(m,4H),2.15–2.02(m,2H),1.70(d,J=6.6Hz,3H).

[0034] Example 5

[0035]

[0036] Following the synthetic method of compound I-1, (R)-3-(1-aminoethyl)-5-(trifluoromethyl)aniline was replaced with (R)-1-(3-fluorophenyl)ethylamine to obtain compound I-5 (32.2 mg, yield 36%). 1 H NMR(300MHz, CDCl3)δ8.53(s,1H),7.39–7.11(m,4H),7.04–6.91(m,2H),5.78–5.59(m,2H),4.16(t,J=6.6Hz,2H), 3.97(s,3H),3.82–3.62(m,4H),2.58(t,J=7.0Hz,2H),2.54–2.41(m,4H),2.19–2.05(m,2H),1.69(d,J=6.5Hz,3H).

[0037] Example 6

[0038]

[0039] Following the synthetic method of compound I-1, by replacing (R)-3-(1-aminoethyl)-5-(trifluoromethyl)aniline with (αR)-1,1-difluoro-2,3-dihydro-α-methyl-1H-ethylene-4-methylamine, compound I-6 (67.2 mg, yield 73%) can be obtained. 1H NMR (300MHz, CDCl3) δ8.53(s,1H),7.58(d,J=7.5Hz,1H),7.52(d,J=7.4Hz,1 H),7.40(dd,J=7.5,7.4Hz,1H),7.19(s,1H),6.87(s,1H),5.79–5.67(m,1H), 5.44(d,J=7.1Hz,1H),4.17(t,J=6.6Hz,2H),3.77–3.69(m,4H),3.23–3.01( m,2H),2.69–2.54(m,4H),2.53–2.45(m,4H),2.19–2.05(m,2H),1.70(s,3H).

[0040] Example 7

[0041]

[0042] Following the synthetic method of compound I-1, replacing (R)-3-(1-aminoethyl)-5-(trifluoromethyl)aniline with (R)-3-(1-aminoethyl)-2-methylbenzyl nitrile, compound I-7 (55.1 mg, yield 72%) can be prepared. 1 H NMR (400MHz, CDCl3) δ8.49 (s, 1H), 7.64 (d, J = 7.8Hz, 1H), 7.53 (d, J = 7.5Hz, 1H), 7.26(dd,J=7.8,7.5Hz,1H),7.18(s,1H),6.96(s,1H),5.81–5.70(m,1H),5.62(d ,J=6.3Hz,1H),4.18(t,J=6.5Hz,2H),3.96(s,3H),3.80–3.68(m,4H),2.70(s,3 H),2.59(t,J=7.0Hz,2H),2.50(s,3H),2.17–2.06(m,2H),1.65(d,J=6.8Hz,3H).

[0043] Example 8

[0044]

[0045] Following the synthetic method of compound I-1, replacing (R)-3-(1-aminoethyl)-5-(trifluoromethyl)aniline with (1S)-1-(6-methyl(2-pyridyl))-ethylamine, compound I-8 (20.8 mg, yield 35%) can be prepared. 1H NMR(300MHz, CDCl3)δ8.56(s,1H),7.69–7.52(m,2H),7.24–7.13(m,3H),7.10(d,J=7.9Hz,1H),5.58–5.45(m,1H),4.25(t,J =7.3Hz,2H),3.99(s,3H),3.82–3.65(m,4H),2.69–2.56(m,5H),2.56–2.41(s,4H),2.24–2.09(m,2H),1.64(d,J=6.7Hz,3H).

[0046] Example 9

[0047]

[0048] Following the synthetic method of compound I-1, replacing (R)-3-(1-aminoethyl)-5-(trifluoromethyl)aniline with (1R)-1-(2-methyl(4-pyridyl))-ethylamine, compound I-9 (97.8 mg, yield 32%) can be obtained. 1 H NMR (300MHz, CDCl3) δ8.49(s,1H),8.43(d,J=5.1Hz,1H),7.20(s,2H),7.17–7.12(m,1H),7.11(s,1H),5.91(d,J=6.5Hz,1H),5.64–5.51(m,1 H),4.20(t,J=6.6Hz,2H),3.98(s,3H),3.83–3.70(m,4H),2.64(t,J=7.0Hz,2H),2.60–2.47(m,5H),2.21–2.02(m,4H),1.68(d,J=7.0Hz,3H).

[0049] Example 10

[0050] In vitro SOS1 protein level activity assay

[0051] The compounds in the above embodiments inhibit SOS1 and KRAS G12C The binding activity was tested, and the specific procedures are as follows:

[0052] 1. SOS1-KRAS G12C Protein inhibitory activity test procedure

[0053] (1) Dissolve the compound to be tested in DMSO to prepare a 10 mM stock solution. Transfer the stock solution to the test plate and perform a concentration gradient dilution.

[0054] (2) Prepare Tag1-SOS1 solution with dilution buffer, transfer 5 μL of Tag1-SOS1 solution to the detection plate, and add 5 μL of dilution buffer to the low blank control group.

[0055] (3) Prepare Tag2-KRAS using dilution buffer. G12C Transfer 5 μL of Tag2-KRAS solution. G12C Pour the solution onto the test plate and add 5 μL of dilution buffer to the low blank control group.

[0056] (4) Prepare Anti-Tag1-Tb using detection buffer. 3+ Prepare the Anti-Tag2-XL665 detection solution. Transfer 10 μL of the detection solution to the detection plate. Place the detection plate in a centrifuge and centrifuge at 1000 rpm for 1 min, then incubate at room temperature for 30 min.

[0057] 2. Data Analysis

[0058] Calculate the ratio of RFU 665nm / RFU 615nm and convert the ratio to a percentage suppression value. Fit the data to Excel to obtain the suppression value using the following formula:

[0059] Inh%=(Max–Signal) / (Max–Min)×100

[0060] Using XL-Fit to fit the data, the half-inhibitory concentration was obtained using the following formula:

[0061] Y=Bottom+(Top–Bottom) / (1+(IC 50 / X)×HillSlope)

[0062] Y is the inhibition rate, and X is the compound concentration.

[0063] 3. Experimental Results

[0064] The specific results are shown in the table below:

[0065]

[0066]

[0067] Example 11

[0068] In vitro EGFR protein level activity assay

[0069] The EGFR enzyme activity of the compounds in the above examples was tested, and the specific procedures were as follows:

[0070] 1. EGFR protein inhibitory activity assay procedure

[0071] (1) Dissolve the test compound in DMSO to prepare a 10 mM stock solution. Perform a concentration gradient dilution and transfer 5 μL of the test compound solution to the detection plate.

[0072] (2) Prepare the kinase solution with dilution buffer, transfer 10 μL of the kinase solution to the detection plate, and add 10 μL of dilution buffer to the low blank control group. Incubate at room temperature for 10 minutes.

[0073] (3) Prepare substrate peptide and ATP solutions using dilution buffer. Transfer 10 μL of substrate peptide and ATP solution to the detection plate, and add 10 μL of dilution buffer to the low blank control group. Incubate at 28°C for a period of time, and then add 25 μL of reaction stop solution to stop the reaction.

[0074] (4) Add 50 μL of chemiluminescence kinase activity detection reagent and use a multi-functional microplate reader to detect the chemiluminescence value of the system.

[0075] 2. Data Analysis

[0076] Fit the data to Excel to obtain the suppression value using the following formula:

[0077] Inh%=(Max–Signal) / (Max–Min)×100

[0078] Using XL-Fit to fit the data, the half-inhibitory concentration was obtained using the following formula:

[0079] Y=Bottom+(Top–Bottom) / (1+(IC 50 / X)×HillSlope)

[0080] Y is the inhibition rate, and X is the compound concentration.

[0081] 3. Experimental Results

[0082] The specific results are shown in the table below:

[0083] Compound numbering <![CDATA[EGFRIC 50 (nM)]]> Compound numbering <![CDATA[EGFRIC 50 (nM)]]> I-1 7855 I-6 11.2 I-2 14.6 I-7 37.5 I-3 8.0 I-8 / 1-4 2.3 I-9 / I-5 2.8 .

[0084] Example 12

[0085] DLD-1(3D) cell proliferation inhibition activity test

[0086] 1. Experimental Procedure

[0087] Healthy DLD-1 colon cancer cells were seeded at a density of 500 cells / well in 96-well plates and cultured overnight at 37°C in a 5% CO2 incubator. Serially diluted compounds were added, and the cells were cultured for another 7 days. Then, 100 μL of the assay kit (Celltiter Glo Assay kit-3D) was added to each well. After shaking for 5 minutes at room temperature, the cells were incubated in the dark at room temperature for 30 minutes. The luminescence values ​​were then read using a multi-mode microplate reader.

[0088] 2. Data Analysis

[0089] The percentage inhibition rate can be calculated using the following formula: Inhibition percentage = 100% × (0% inhibition group signal value – signal value of the analyte at a specific concentration) / (0% inhibition group signal value – 100% inhibition group signal value).

[0090] Compound IC 50 The values ​​were fitted using XL-Fit data and calculated using the following formula: Y = Bottom + (Top –

[0091] Bottom) / (1+(IC 50 / X)×HillSlope), where Y is the inhibition rate and X is the compound concentration.

[0092] The specific results are shown in the table below:

[0093]

[0094]

Claims

1. A quinazoline amine compound as shown in the general formula I and a pharmaceutically acceptable salt thereof, with the structure as follows: Ar is selected from or ; wherein, R 1 - R 3 are each independently selected from hydrogen, deuterium, halogen, methyl, trifluoromethyl, cyano or amino.

2. A quinazoline amine compound as shown in formula I according to claim 1 and pharmaceutically acceptable salts thereof, characterized in that, ###0001### Formula I The compound as shown in the formula I is any one of the following compounds: 。 3.A pharmaceutical composition comprising the compound of claims 1-2 and a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 4.The use of the quinazoline amine compound and the pharmaceutically acceptable salt thereof according to claims 1-2, and the pharmaceutical composition according to claim 3 for preparing a SOS1 / EGFR dual-target inhibitor.

5. The use of the quinazolinamine compound or pharmaceutically acceptable salt thereof according to claim 1 to 2, or the pharmaceutical composition according to claim 3, in the manufacture of a medicament for the prophylaxis or treatment of cancer, characterized in that, The cancer is colon cancer.

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