2-arylamino pyrimidines and their use in the treatment of breast cancer

Abstract

This invention relates to the fields of pharmaceuticals and chemicals, specifically to 2-arylaminopyrimidine compounds and their use in the treatment of breast cancer, including methods for preparing 2-arylaminopyrimidine compounds and their application in the preparation of drugs for treating breast cancer. The invention provides 2-arylaminopyrimidine compounds of general formula (I) or pharmaceutically acceptable salts thereof, wherein Ar is as described in the claims and specification. The 2-arylaminopyrimidine compounds of the present invention, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, can inhibit FAK tyrosine kinase activity, inhibit tumor proliferation, and can be used to prepare FAK tyrosine kinase inhibitors, as well as antitumor drugs, particularly for the preparation of drugs against breast cancer.

Description

Technical Field

[0001] This invention relates to the fields of pharmaceuticals and chemicals, specifically to 2-arylaminopyrimidine compounds and their application in the treatment of breast cancer, including methods for preparing 2-arylaminopyrimidine compounds and their use in the preparation of drugs for treating breast cancer. Background Technology

[0002] Focal adhesion kinase (FAK) is a non-receptor protein tyrosine kinase that acts as a key adaptor protein in integrin-activated signaling, initiating downstream signaling cascades to regulate various biological functions of cancer cells. Numerous studies have shown that FAK-mediated signaling pathways regulate tumor cell survival, proliferation, invasion, adhesion, and migration through both kinase-dependent and kinase-independent mechanisms, thereby promoting tumor progression and metastasis. Research has found elevated FAK mRNA levels in approximately 26% of breast cancer tumors, with FAK protein overexpression being particularly common in human epidermal growth factor receptor 2 (HER2)-positive and triple-negative breast cancer. FAK overexpression is significantly associated with accelerated tumor proliferation, enhanced invasiveness, elevated histological grade, and poor patient prognosis.

[0003] Studies using mouse tumor models have shown that FAK gene knockout can effectively inhibit multiple stages of breast cancer development and progression. Research on FAK inactivation through gene intervention or drug inhibition has confirmed that the FAK signaling pathway is involved in promoting tumor growth, angiogenesis, and metastasis. In vitro experiments have shown that sub-micromolar concentrations of FAK inhibitors can block tumor cell proliferation under three-dimensional culture conditions; this phenomenon is closely related to the low expression of the tumor suppressor protein merlin and the activation of the β1 / β5 integrin signaling pathway in tumor cells. Notably, the survival of tumor spheroids is highly dependent on FAK activity.

[0004] Breast cancer is one of the most common malignant tumors among women worldwide. As a highly heterogeneous disease, its response to treatment varies significantly. Given the urgent need for effective treatment of breast cancer, it is essential to develop anti-breast cancer drugs with unique mechanisms of action and novel structures to expand the range of drugs available for breast cancer treatment. Summary of the Invention

[0005] One of the objectives of this invention is to provide the 2-arylaminopyrimidine compounds or pharmaceutically acceptable salts thereof, which have good anti-breast cancer activity.

[0006] Another object of the present invention is to provide a pharmaceutical composition comprising the 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof.

[0007] A third object of the present invention is to provide the use of the 2-arylaminopyrimidine compounds or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof in the preparation of FAK tyrosine kinase inhibitors.

[0008] A fourth object of the present invention is to provide the use of the 2-arylaminopyrimidine compounds or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof in the preparation of medicaments for treating breast cancer.

[0009] This invention is achieved through the following technical solution:

[0010] A 2-arylaminopyrimidine compound of general formula (I) or a pharmaceutically acceptable salt thereof:

[0011]

[0012] Wherein, Ar is a substituted phenyl group, and the substituent is a halogen, C1-C6 alkoxy, C1-C6 alkyl, or -HN-(CH2). m CO-R, -O(CH2) n -R, a six-membered heterocyclic group, wherein the heterocyclic group contains 1-3 N, O or S heteroatoms;

[0013] The heterocyclic group is replaced by one or more H, hydroxyl, C1-C6 alkyl, C1-C6 alkyl groups substituted with hydroxyl, or -CO-(CH2). p CH3 substitution;

[0014] R is ;

[0015] m, n, p = 0-3;

[0016] Furthermore,

[0017] The six-membered heterocyclic group contains 1-2 N or O heteroatoms;

[0018] The heterocyclic group can be substituted by one or more H, C1-C4 alkyl, hydroxyl, C1-C4 alkyl group substituted with hydroxyl, or COCH3;

[0019] Ar represents a substituted phenyl group, wherein the substituent is a halogen, C1-C4 alkoxy, C1-C4 alkyl, or -HN-(CH2). m CO-R, -O(CH2) n -R, a six-membered heterocyclic group, wherein the heterocyclic group is:

[0020] , , ;

[0021] Furthermore, Ar is selected from:

[0022] .

[0023] Specifically, the present invention preferably uses the following 2-arylaminopyrimidine compounds or pharmaceutically acceptable salts thereof:

[0024]

[0025] The compounds of the present invention are bases, wherein the desired salt form can be prepared by suitable methods known in the art, including treating the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or treating the free base with an organic acid such as acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranoside (e.g., glucuronic acid or galacturonic acid), α-hydroxy acid (e.g., citric acid or tartaric acid), amino acid (e.g., aspartic acid or glutamic acid), aromatic acid (e.g., benzoic acid or cinnamic acid), sulfonic acid (e.g., toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid), etc. The pharmaceutically acceptable salts described in this invention include sulfates, pyrosulfates, bisulfates, sulfites, phosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, octanoates, acrylates, formates, isobutyrates, hexanoates, heptaates, propionates, oxalates, malonates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, γ-hydroxybutyrate, glycolic acid salts, tartrates, amygdalinates and sulfonates, xylenesulfonates, methanesulfonates, propanesulfonates, naphthalene-1-sulfonates and naphthalene-2-sulfonates.

[0026] The present invention also provides a method for preparing the 2-arylaminopyrimidine compound represented by general formula (I) or a pharmaceutically acceptable salt thereof, the reaction process of which is as follows:

[0027]

[0028] Ar is as previously mentioned.

[0029] The pharmaceutical compositions of the present invention typically contain one of the compounds of the present invention. However, in some embodiments, the pharmaceutical compositions of the present invention contain one or more of the compounds of the present invention.

[0030] In addition, the pharmaceutical composition of the present invention may also be a composition of one or more compounds of the present invention and one or more other pharmaceutically active compounds.

[0031] This invention provides the use of the 2-arylaminopyrimidine compounds or pharmaceutically acceptable salts thereof in the preparation of FAK tyrosine kinase inhibitors.

[0032] The present invention provides the use of pharmaceutical compositions containing the 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof in the preparation of FAK tyrosine kinase inhibitors.

[0033] The 2-arylaminopyrimidine compounds of the present invention, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, can inhibit tumor proliferation and can be used to prepare antitumor drugs. The tumor is preferably breast cancer.

[0034] The compounds described in this invention, or pharmaceutically acceptable salts thereof, or combinations thereof, exert their anti-breast cancer effects by inhibiting FAK tyrosine kinase.

[0035] The compounds of this invention have the potential to be developed into novel and highly effective FAK inhibitors. Screening for antitumor activity shows that the compounds of this invention have a strong ability to inhibit the proliferation of breast cancer cells MCF-7, SKBR-3, and HK-2, and have significant application value for the treatment of related tumor diseases, especially breast cancer. Attached Figure Description

[0036] Figure 1 Experiments on xenografting of SKBR-3 breast cancer cells with compound I-7;

[0037] A: Tumor volume change; B: Mouse weight change; C: Tumor photograph; D: Tumor weight; E: Tumor inhibition rate. Detailed Implementation

[0038] The present invention will be further described and explained below with reference to specific embodiments, but these embodiments are not intended to limit the scope of the present invention.

[0039] Experimental methods in this invention, where specific conditions are not specified, are generally performed under conventional conditions or as recommended by the raw material or product manufacturer. Reagents whose specific source is not specified are commercially available, conventional reagents.

[0040] Preparation process of compounds I-1 to I-11

[0041]

[0042] Example 1: Preparation of compounds I-1 to I-11

[0043] Compounds I-1 to I-11 were prepared according to the reaction procedure described above.

[0044] Compound 1 (1.0 eq.) and DIPEA (3.0 eq.) were in THF, 0o Slowly add 2 (1.0 eq.) of hydrochloride under the following conditions: C. After addition, heat to 40°C. o The reaction was carried out at C for 1 h, and TLC monitoring showed that the reaction was complete. After evaporating THF, the mixture was extracted with DCM / water, and the organic layer was dried with anhydrous sodium sulfate. After removing the solvent under reduced pressure, 3 was obtained. 3 was placed in DCM, TFA was added, and the mixture was stirred overnight at room temperature. After slowly adding saturated sodium bicarbonate solution to neutralize the TFA, the organic layer was dried with anhydrous sodium sulfate, and column chromatography was used to separate 4. 4 (1.0 eq.), imidazole (2.0 eq.) was placed in THF, TBDMSOTf (2.0 eq.) was added, and the mixture was stirred overnight at room temperature. The reaction was carried out at TLC monitoring that the reaction was complete. After removing the organic solvent under reduced pressure, the mixture was extracted with DCM / water, and the organic layer was dried with anhydrous sodium sulfate. Column chromatography was used to separate 5. 5a–b (1.0 eq.), ArNH2 (1.0 eq.), Pd2(dba)3 (0.1 eq.), RuPhos (0.1 eq.), Cs2CO3 (2.5 eq.) was placed in 1,4-dioxane, 100 o After reacting at C for 12 h, the solid matter was filtered off, the organic solvent was evaporated, and the mixture was extracted with DCM / water. The organic layer was dried over anhydrous sodium sulfate and separated by column chromatography to obtain 6a–k. 6a–k (1.0 eq.) and diethylamine (2.0 eq.) were added to THF, and TBAF (4.0 eq.) was added dropwise. The reaction was carried out for 5 min, the THF was evaporated, and the mixture was separated by column chromatography (DCM:MeOH) to purify the target compounds I-1 to I-11.

[0045] Compound I-1:

[0046] 1 H NMR (CDCl3, 600 MHz) d 8.86 (1H, bro. s), 8.09 (1H, d, J = 8.7 Hz), 7.99(1H, s), 7.84 (1H, d, J = 7.4 Hz), 7.70 (1H, d, J = 7.3 Hz), 7.54 (1H, t, J = 7.6Hz), 7.09 (1H, s), 6.57 (1H, s), 6.50 (1H, s), 6.44 (1H, d, J = 8.6 Hz), 6.16(1H, t, J = 5.0 Hz), 4.91 (2H, d, J = 5.4 Hz), 3.86 (7H, bro. s), 3.09 (4H, t,J = 4.1 Hz), 3.00 (3H, s). 13 C NMR(CDCl3, 101 MHz) d 156.6, 156.0, 150.0, 149.5, 147.0, 141.1, 140.9, 132.5, 129.8, 126.3, 126.2, 122.6, 120.4, 115.0, 107.9, 103.7, 100.3, 95.2, 67.0 (2C), 55.7, 50.5 (2C), 44.5, 44.1。

[0047] Compound I-2:

[0048] 1 H NMR(DMSO- d 6, 400 MHz) d 11.31 (1H, bro. s), 7.97 (1H, s), 7.90 (1H, d, J = 8.7 Hz), 7.79 (1H, d, J = 7.6 Hz), 7.70 (1H, d, J = 7.6 Hz), 7.59 (1H, t, J = 7.7 Hz), 7.21 (1H, d, J = 6.6 Hz), 7.08 (1H, s), 6.93 (1H, d, J = 1.8 Hz), 6.58 (1H, s), 6.38 (1H, d, J = 8.6 Hz), 4.77 (2H, d, J = 6.0 Hz), 4.65 (1H, d, J = 4.1 Hz), 3.80 (3H, s), 3.61 - 3.56 (1H, m), 3.45 - 3.42 (2H, m), 3.16 (3H, s), 2.02 - 1.96 (2H, m), 1.83 - 1.80 (2H, m). 13 C NMR(DMSO- d 6, 101 MHz) d157.0, 156.0, 151.5,149.5, 147.2, 142.7, 141.2, 132.8, 129.8, 126.0, 125.6, 122.3, 120.7, 116.1,107.9, 101.9, 101.1, 94.9, 66.6, 56.1, 48.1 (2C), 44.0, 43.6, 34.6 (2C).

[0049] Compound I-3:

[0050] 1 H NMR (DMSO- d 6,400 MHz) d 11.26 (1H, bro. s), 8.50 (1H, s), 7.98 (1H, s), 7.79 (1H, d, J = 7.5 Hz), 7.74 (1H, d, J = 7.5 Hz), 7.60 (1H, t, J = 7.6 Hz), 7.46 (2H, d, J = 6.6 Hz), 7.18 (1H, t, J = 5.5 Hz), 6.93 (1H, s), 6.77 (2H, d, J =6.6 Hz), 4.81 (2H, d, J = 6.0 Hz), 4.65 (1H, bro. s), 3.58 (1H, bro. s), 3.43-3.36 (2H, m), 3.16 (3H, s), 2.70 (2H, t, J = 10.1 Hz), 2.03-1.97 (2H, m), 1.83-1.81 (2H, m). 13 C NMR (DMSO- d 6, 101 MHz) d 157.1, 156.0, 151.5, 146.0, 142.8,141.2, 134.1, 132.7, 129.8, 125.9, 125.6, 120.0 (2C), 116.9 (2C), 115.9,101.9, 94.7, 66.6, 48.2 (2C), 44.0, 43.5, 34.6 (2C).

[0051] Compound I-4:

[0052] 1 ¹H NMR (DMSO- d d6, 400 MHz) d 11.3 (1H, bro. s), 8.52 (1H, s), 7.97 (1H, s), 7.79 (1H, d, J J = 7.7 Hz), 7.74 (1H, d, J J = 7.7 Hz), 7.60 (1H, t, J J = 7.7 Hz), 7.49 (2H, d, J J = 8.2 Hz), 7.18 (1H, t, J J = 5.7 Hz), 6.93 (1H, bro. s), 6.76 (2H, d, J J = 8.2 Hz), 4.80 (2H, d, J J = 6.0 Hz), 3.16 (3H, s), 2.99 (4H, bro. s), 2.52 (4H, bro. s), 2.48 (2H, bro. s), 1.00 (3H, t, J J = 7.2 Hz). 13 ¹³C NMR (DMSO- d d6, 101 MHz) d 157.1, 156.0, 151.6, 145.7, 142.8, 141.2, 134.5, 132.7, 129.8, 125.9, 125.6, 120.0 (2C), 116.4 (2C), 115.9, 101.9, 94.7, 52.8 (2C), 52. (2C), 49.4 (2C), 44.0, 43.5, 12.1.

[0053] Compound I-5:

[0054] 1 ¹H NMR (CDCl3, 600 MHz) d 10.25 (1H, s), 8.00 (1H, s), 7.83 (1H, d, J J = 7.8 Hz), 7.67 (1H, d, J J = 7.7 Hz), 7.53 (1H, t, J J = 7.8 Hz), 7.37 (2H, d,[[ID=4S]] J= 8.7 Hz), 6.85 (2H, d, J = 8.8 Hz), 6.67 (1H, s), 6.13 (1H, t, J = 6.0 Hz), 6.04 (1H, s), 4.85 (1H, d, J = 5.9 Hz), 3.75 (2H, t, J = 5.0 Hz), 3.60 (2H, t, J = 5.0 Hz), 3.09 (2H, t, J = 5.0 Hz), 3.05 (2H, t, J = 5.0 Hz), 3.02 (3H, s), 2.12 (3H, s). 1 1H NMR (CDCl3, 101 MHz) d 169.1, 156.6, 156.6, 150.2, 146.9, 141.2, 140.8, 133.1, 132.6, 129.7, 126.5, 126.1, 121.9 (2C), 117.8 (2C), 115.3, 102.9, 95.3, 50.5, 50.2, 46.3, 44.5, 43.9, 41.4, 21.4.

[0055] Compound I-6:

[0056] 1 1H NMR (DMSO- d d6, 400 MHz) d 11.2 (1H, d, = 9.0 Hz), 7.18 (1H, t, J = 2.4 Hz), 8.53 (1H, s), 7.98(1H, s), 7.79 (1H, d, J = 7.7 Hz), 7.74 (1H, d, J = 7.7 Hz), 7.60 (1H, t, J J = 6.0 Hz), 6.93 (1H, d, J = 2.4 Hz), 6.76 (2H, d, J = 2.4 Hz), 6.76 (2H, d, J = 9.0 Hz), 4.80 (2H, d, J= 6.0 Hz), 3.72 (4H, t, J = 4.4 Hz), 3.16(3H, s), 2.99 (4H, t, J = 4.5 Hz). 13 C NMR(DMSO- d 6, 101 MHz) d 157.0, 156.0, 151.6,145.7, 142.8, 141.2, 134.6, 132.7, 129.8, 125.9, 125.6, 120.0 (2C), 116.1(2C), 115.9, 101.9, 94.7, 66.7 (2C), 50.0 (2C), 44.0, 43.5.

[0057] Compound I-7:

[0058] 1 H NMR(DMSO- d 6, 400 MHz) d 11.40 (1H, bro. s), 8.85 (1H, s), 7.97 (1H,s), 7.79 (1H, d, J = 7.6 Hz), 7.75-7.69 (2H, m), 7.60 (1H, t, J = 7.8 Hz), 7.28-7.22 (2H, m), 6.99 (1H, s), 6.86 (1H, t, J = 9.0 Hz), 4.80 (2H, d,​​​​​​​​​​​​​​​= 9.1 Hz), 132.7, 129.8, 125.8, 125.6, 119.8 ( J = 4.0 Hz), 116.3,114.4 ( J = 2.0 Hz), 106.6 ( J = 26.0 Hz), 101.9, 95.0, 66.3, 49.4 (2C), 43.9,43.4, 35.1 (2C).

[0059] Compound I-8:

[0060] 1 H NMR(DMSO- d 6, 400 MHz) d 11.40 (1H, d, J = 2.4 Hz), 8.85 (1H, s), 7.97(1H, s), 7.80 - 7.70 (3H, m), 7.60 (1H, t, J = 7.8 Hz), 7.30 (1H, dd, J = 8.7, 1.8Hz), 7.25 (1H, t, J = 6.2 Hz), 6.99 (1H, d, J = 2.4 Hz), 6.86 (1H, t, J = 9.4 Hz),4.80 (2H, d, J = 6.0 Hz), 3.16 (3H, s), 2.96 (4H, bro. s), 2.69 - 2.54 (4H, m),2.31 (3H, s). 13 C NMR(DMSO- d 6, 101 MHz) d 156.6, 156.1, 154.4 ( J = 265.0 Hz),151.1, 142.6, 141.2, 137.3 ( J = 11.0 Hz), 133.2 ( J = 9.1 Hz), 132.7, 129.8,125.9, 125.6, 119.6 ( J = 4.0 Hz), 116.4, 114.3 ( J = 2.0 Hz), 106.6 ( J= 26.0Hz), 101.9, 95.1, 55.4 (2C), 50.4 (2C), 44.0, 43.5.

[0061] Compound I-9:

[0062] 1 H NMR (DMSO- d 6,400 MHz) d 11.40 (1H, bro. s), 8.85 (1H, s), 7.97 (1H,s), 7.80-7.70 (3H, m), 7.60 (1H, t, J = 7.8 Hz), 7.31-7.24 (2H, m), 6.99 (1H,s), 6.87 (1H, t, J = 8.8 Hz), 4.80 (2H, d, J = 6.0 Hz), 3.60 (2H, bro. s), 3.16(3H, s), 2.98 (4H, bro. s), 2.52 (6H, bro. s). 13 C NMR (DMSO- d 6, 101 MHz) d 156.5, 156.1, 154.4 ( J = 264.0 Hz), 151.2, 142.6, 141.2, 137.6 ( J = 11.0 Hz), 133.5 ( J = 9.1 Hz), 132.7, 129.8, 125.9, 125.6, 119.5 ( J = 4.0 Hz), 116.4, 114.4 ( J = 2.0 Hz), 106.7 ( J = 26.0 Hz), 101.9, 95.1, 60.6, 55.4, 53.7 (2C), 50.9 (2C), 44.0, 43.5.

[0063] Compound I-10:

[0064] 1 H NMR (DMSO- d 6,400 MHz) d11.32 (1H, bro. s), 9.43 (1H, s), 8.74 (1H,s), 7.97 (1H, s), 7.79 (1H, d, J J = 7.8 Hz), 7.75 (1H, d, J J = 7.8 Hz), 7.64 - 7.56(3H, m), 7.39 (2H, d, J J = 8.7 Hz), 7.21 (1H, t, J J = 5.8 Hz), 6.96 (1H, d, J J = 2.0Hz), 5.32 (1H, t, J J = 4.8 Hz), 4.83 (2H, d, J J = 6.0 Hz), 4.60 (1H, bro. s), 3.51(1H, bro. s), 3.30 (2H, bro. s), 3.16 (3H, s), 3.10 - 3.01 (2H, m), 2.81 - 2.69(2H, m), 2.02 - 1.96 (2H, m), 1.80 - 1.72 (2H, m). 13 C NMR(DMSO - d d6, 101 MHz) d 156.8, 156.1, 151.4, 142.7, 141.2, 137.8, 132.8, 131.8, 130.1, 129.8, 125.9,125.6, 120.3 (2C), 118.9 (2C), 116.2, 101.9, 94.9, 66.2, 52.3 (2C), 43.9,43.5, 40.0 (2C), 34.6 (2C).

[0065] Compound I - 11:

[0066] 1 H NMR(DMSO - d d6, 400 MHz) d 11.32 (1H, bro. s), 8.62 (1H, s), 7.79 (1H,d, J J = 7.8 Hz), 7.74 (1H, d, J J = 7.8 Hz), 7.61 (1H, t, J J = 7.7 Hz), 7.54 (2H, d, J=8.7 Hz), 7.21 (1H, t, J = 5.8 Hz), 6.94 (1H, d, J = 2.0 Hz), 6.76 (2H, d, J = 8.7Hz), 5.02 (1H, s), 4.82 (2H, d, J = 6.0 Hz), 3.97 (3H, t, J = 5.7 Hz), 3.16 (3H,s), 3.14-2.90 (6H, m), 2.17-2.08 (2H, m), 2.03-1.97 (2H, m), 1.74-1.65 (2H,m). 13 C NMR (DMSO- d 6, 101 MHz) d 157.0, 156.0, 152.8, 151.5, 142.7, 141.2, 135.5,132.7, 129.8, 125.8, 125.6, 120.2 (2C), 116.0, 114.7 (2C), 101.9, 94.8, 70.2, 65.9 (2C), 53.8 (3C), 50.6 (2C), 44.0, 43.5, 35.6, 34.9, 27.0.

[0067] Example 2: Inhibitory activity of the compound against FAK kinase and breast cancer cells

[0068] (1) FAK kinase activity assay

[0069] a) Prepare a 2×ATP / substrate solution and a 2×kinase solution using kinase reaction buffer.

[0070] b) Using an Echo 655, transfer 100 nL of the compound dilution to a 384-well plate; after centrifugation, add 5 μL of 2× kinase solution to the plate, centrifuge at 1000 rpm for 1 minute, and then at 25°C. o Incubate at C for 10 minutes.

[0071] c) Add 5 μL of 2× substrate and ATP solution to a 384-well plate, centrifuge at 1000 rpm for 1 minute, and then at 25°C. o Incubate at C for 60 minutes.

[0072] d) Prepare 2×XL665 and antibody detection reagents using detection buffer.

[0073] e) Add 10 μL of kinase assay reagent to the assay plate and incubate at 25°C. o Incubate at C. Centrifuge at 1000 rpm for 1 minute, then incubate at 25°C. o Continue incubation at C for 1 hour.

[0074] f) Read the fluorescence signals at 620 nm (Cryptate) and 665 nm (XL665) on the BMG instrument.

[0075] (2) Cell viability detection

[0076] (1) Cell types: BAF3-PTK2 cells, SKBR-3 breast cancer cells.

[0077] (2) Cell seeding: Collect cells in the logarithmic growth phase and adjust the cell suspension concentration to 4 × 10⁴ cells per well. 3 100 μL of cells were seeded into each well of a 96-well plate, with 3 replicates per group (margin wells were filled with sterile PBS).

[0078] (3) Cell culture: After cell seeding, the control group was cultured with 10% FBS RPMI-1640 (SKBR-3) and HyCyte®TCM-G786 (BAF3-PTK2), while the experimental groups were cultured with different concentration gradients of the target compound and 37% FBS RPMI-1640 (SKBR-3) and HyCyte®TCM-G786 (BAF3-PTK2), respectively. o Continue culturing in a 5% CO2 incubator for 72 h.

[0079] (4) Color development: 10 μL of CCK-8 solution (5 mg / ml) was added to each of the two groups of cells after 72 h of culture, and the culture was terminated after 4 h.

[0080] (5) Colorimetric analysis: The absorbance value (OD value) of each well was measured on an enzyme-linked immunosorbent assay (ELISA) instrument. A wavelength of 450 nm was selected, and the blank wells of cell-free RPMI-1640 culture medium were used to zero the absorbance value of each well.

[0081] (6) Record the results: Cell growth inhibition rate = (absorbance value of control group - absorbance value of experimental group) / absorbance value of control group × 100%, cell proliferation rate = (absorbance value of experimental group / absorbance value of control group) × 100%.

[0082] (7) Plotting cell growth curves: Plotting the inhibitor concentration using the GraphPad Prism plotting software within GraphPad software, and estimating the IC50. 50 The results are shown in Table 1.

[0083]

[0084] Example 3: Results of xenograft experiment on SKBR-3 breast cancer cells using compound I-7

[0085] After digestion, healthy SKBR-3 cells were resuspended in PBS and stored on ice. 100 μL of the cell suspension was subcutaneously injected into the right axilla of a mouse (NUNU). Tumor volume was measured 7 days later; the tumor had grown to approximately 100 mm. 3 At that time, the tumor-bearing mouse model was successfully established.

[0086] Experimental groups: control group, TAE226 group, and compound I-7 group (high and low).

[0087] Dosage regimen:

[0088] (1) Control group: physiological saline, orally.

[0089] (2) TAE226 group (positive control group): 30 mg / kg, administered by gavage once daily.

[0090] (3) Group I-7: 30 or 60 mg / kg administered by gavage once daily.

[0091] Results determination method: Tumor size was measured every other day;

[0092] V = 0.5 × a × b^2 (V is the tumor volume, a is the long diameter of the tumor, and b is the short diameter of the tumor).

[0093] The results are as follows Figure 1 As shown, the results indicate that the tumor growth rate was the fastest in the control group during treatment, while the growth rate in the TAE226 group was slower compared to the saline group. At the same dose, I-7 could significantly inhibit tumor growth, and the inhibition rate was further enhanced when the dosage was increased to 60 mg / kg.

Claims

1. A 2-arylaminopyrimidine compound of general formula (I) or a pharmaceutically acceptable salt thereof, ； The structures of the 2-arylaminopyrimidine compounds or their pharmaceutically acceptable salts are as follows: 。 2. The method for preparing the 2-arylaminopyrimidine compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The reaction process is as follows: ； Ar is as described in claim 1.

3. A pharmaceutical composition comprising the 2-arylaminopyrimidine compound of claim 1 or a pharmaceutically acceptable salt thereof.

4. A pharmaceutical composition comprising the 2-arylaminopyrimidine compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3 and other antitumor drugs.

5. The use of the 2-arylaminopyrimidine compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3 or 4, in the preparation of a FAK tyrosine kinase inhibitor.

6. The use of the following 2-arylaminopyrimidine compounds or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof in the preparation of anti-breast cancer drugs: 。

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