WO2020001351A1 - Egfr inhibitor, method for preparing the same, and uses thereof - Google Patents

Abstract

Provided are a 2-aminopyrimidine derivative having a structure shown in a formula (I), a pharmaceutical composition containing the compound of formula (I), and the uses thereof for preparing an epidermal growth factor receptor (EGFR) kinase for preventing or treating diseases related thereto, especially uses for preventing or treating cancers related to the EGFR.

Description

EGFR inhibitor and its preparation and application Technical field

The present application belongs to the technical field of medicine, and particularly relates to a 2-aminopyrimidine derivative and use thereof for preparing an antitumor drug.

This application claims the priority of Chinese patent CN201810704813.7 (filed on June 27, 2018, the invention name EGFR inhibitor and its preparation and application).

Background technique

Epidermal growth factor receptor (EpidermalRowthFactorReceptor, EGFR) is a transmembrane receptor protein with tyrosine kinase activity widely distributed on human tissue cell membranes, and is a member of the erbB receptor family of tyrosine kinases. By binding to the ligand epidermal growth factor (EGF), EGFR forms a homodimer on the cell membrane, or forms a heterodimer with other receptors in the family (such as erbB2, erbB3, or erbB4) to activate and cause Phosphorylation of key tyrosine residues in EGFR cells activates the kinase domain and further activates multiple downstream signaling pathways within the cell. These intracellular signaling pathways play important roles in cell proliferation, survival, and anti-apoptosis.

The tyrosine kinase domain of EGFR can be mutated, resulting in the activation of constitutive signals. This active signaling pathway plays a vital role in the growth, survival and migration of tumor cells. The most common activating mutations are an in-frame deletion mutation in exon 19 and a missense mutation in the 858 codon (L858R). Lung cancer with EGFR mutations is highly sensitive to EGFR tyrosine kinase inhibitors (TKIs) (Science [2004] No. 304, 1497-500). The first generation of tyrosine kinase inhibitors that target EGFR (such as Gefitinib, erlotinib) and other drugs have achieved great success in the clinical treatment of non-small cell lung cancer (New England Journal of Medicine [2004] No. 350, 2129-39; The Lancet Oncology [2012] No. 13 Volume 3, 239-46). However, patients treated with TKI inhibitors often face relapse problems due to the development of resistance. The most common drug resistance mechanism is the second mutation of T790M in EGFR, which is present in about 50% of patients with drug-resistant tumors (PLOS Medicine [2005] No. 2, 1-11). Second-generation EGFR irreversible inhibitors such as Canertinib and Afatinib can overcome drug resistance, but these molecules have poor selectivity for EGFR T790M mutants, and their inhibitory effect on wild-type EGFR is relatively low. Strong, lower tolerated doses in the body.

Therefore, it is necessary to develop a third-generation small molecule EGFR inhibitor with better activity and selectivity, which can selectively inhibit mutants such as T790M, exon 19 deletion, and L858R missense mutation. Lower inhibitory activity to avoid side effects of hyperglycemia caused by off-target (Transl Lung Cancer Res [2015] No. 5, 576-83).

CN105503827A and CN106187915A disclose compounds that selectively inhibit the EGFR T790M mutant. The compounds of the present invention are different from their structures, and the compounds of the present invention are generally superior to CN105503827A in their EGFR kinase and cell inhibitory activity and selectivity to EGFR wild-type / mutant And CN106187915A patent.

Summary of the invention

An object of the present invention is to provide a 2-aminopyrimidine EGFR inhibitor which is highly selective for EGFR mutation and has higher safety.

Another object of the present invention is to provide the use of the EGFR inhibitor in the preparation of a medicament for preventing or treating an epidermal growth factor receptor (EGFR) kinase-related disease.

To achieve the objective of the present invention, the technical solution of the present invention is as follows:

The compound of the present invention represented by the following formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

Wherein R ^{1 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more selected from halogen, hydroxyl, C ₁ -C ₈ alkyl, C ₁ -C ₈ Alkoxy, halogen-substituted C ₁ -C ₈ alkoxy, C ₃ -C ₈ cycloalkyl or C ₃ -C ₈ cycloalkoxy substituted by a substituent;

R ^{2 is} selected from hydrogen, deuterium, halogen, cyano, nitro, C ₁ -C ₈ alkyl, C ₁ -C ₈ alkoxy, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, trifluoromethyl, difluoromethyl or trifluoromethoxy;

R ^{3 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of halogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ alkoxy, Halogen-substituted C ₁ -C ₈ alkoxy, C ₃ -C ₈ cycloalkyl, or C ₃ -C ₈ cycloalkoxy;

X is N or CH;

R ^{4 is} selected from hydrogen, halogen, cyano, nitro, C ₁ -C ₈ alkyl, halo C ₁ -C ₈ alkyl, -C (O) R ⁵ , -C (O) NR ⁵ R ⁶ , -OR ⁵ , -NR ⁵ R ⁶ , -NR ⁵ C (O) R ⁶ , -NR ⁷ (CH ₂ ) _m NR ⁵ R ⁶ , -NC (O) R ⁷ (CH ₂ ) _m NR ⁵ R ⁶ , -NR ⁷ (CH ₂ ) _m NR ⁵ C (O) R ⁶ , -NR ⁷ (CH ₂ ) _m OR ⁶ , -NC (O) R ⁷ (CH ₂ ) _m OR ⁶ , -O (CH ₂ ) _m NR ⁵ R ⁶ or -O (CH ₂ ) _m NR ⁵ C (O) R ⁶ ;

R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, wherein C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl is optional Is further substituted with one or more selected from halogen, hydroxy, -NR ⁸ R ⁹ , C ₁ -C ₈ alkyl, C ₁ -C ₈ alkoxy, halogen substituted C ₁ -C ₈ alkoxy, C ₃ -C ₈ cycloalkyl or C ₃ -C ₈ cycloalkoxy substituents;

Alternatively, R ⁵ , R ⁶ and R ⁷ may independently form a 4-10 membered heterocyclic group;

m is 1, 2, 3 or 4;

R ⁸ and R ⁹ are each independently selected from hydrogen, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl.

According to a preferred embodiment of the present invention, the compound of formula (I), wherein:

R ^{1 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further substituted with one or more substituents selected from halogen or hydroxyl;

R ^{2 is} selected from hydrogen, deuterium, halogen or methyl;

R ^{3 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further substituted with one or more substituents selected from halogen;

X is N or CH;

R ^{4 is} selected from hydrogen, halogen, cyano, nitro, C ₁ -C ₈ alkyl, -NR ⁵ R ⁶ or -NR ⁷ (CH ₂ ) _m NR ⁵ R ⁶ ;

R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

Or R ⁵ , R ⁶ and R ⁷ may independently form a 4-10 membered heterocyclic group;

m is 1, 2, 3 or 4.

According to a preferred embodiment of the present invention, the compound of formula (I) is characterized in that:

R ^{1 is} selected from C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl, optionally further substituted by one or more substituents selected from halogen;

R ^{2 is} selected from hydrogen, deuterium, halogen or methyl;

R ^{3 is} selected from C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl;

X is N or CH;

R ^{4 is} selected from -NR ⁵ R ⁶ or -NR ⁷ (CH ₂ ) _m NR ⁵ R ⁶ ;

R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl;

Or R ⁵ , R ⁶ , and R ⁷ may independently form a 6-8 membered heterocyclic group;

m is 1, 2 or 3.

Further, in a preferred embodiment of the present invention, the compound of formula (I) is characterized in that:

R ^{1 is} selected from methyl, ethyl, difluoromethyl, n-propyl, isopropyl or trifluoroethyl;

R ^{2 is} selected from hydrogen, deuterium, F, Cl, Br or methyl;

R ^{3 is} selected from methyl, ethyl, n-propyl, isopropyl or cyclopropyl;

X is N or CH;

R ⁴ is -NR ⁷ (CH ₂ ) _m NR ⁵ R ⁶ ;

R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl or cyclopropyl;

Or R ⁵ , R ⁶ , and R ⁷ may independently form a 6-membered heterocyclic group;

m is 1, 2 or 3.

As the most preferred embodiment, a particularly preferred compound of the general formula (I) or a pharmaceutically acceptable salt thereof includes the following:

Another aspect of the present invention provides a method for preparing a compound of formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, comprising the following steps:

A compound of formula (II) and a compound of formula (III) undergo a substitution reaction in the presence of a metal catalyst to obtain a compound of formula (IV), and a reduction reaction to obtain a compound of general formula (V), and a condensation reaction to obtain a compound of formula (I), wherein R ¹ , R ² , R ³ , R ⁴ , X are as defined for a compound of formula (I).

Another aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective dose of the aforementioned compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

According to another aspect of the present invention, the aforementioned compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or the aforementioned pharmaceutical composition is prepared for treating EGFR mutants, especially L858R, EGFR mutant, T790M EGFR mutants and deletion of exon 19 activate mutants for the treatment of disease-mediated diseases.

According to another aspect of the present invention, the aforementioned compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or the aforementioned pharmaceutical composition is prepared for treating a disease mediated by EGFR mutant activity alone or in part. Application in therapeutic drugs.

Another aspect of the present invention provides the aforementioned compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or the use of the aforementioned pharmaceutical composition in the preparation of a medicament for treating cancer.

As a further preferred embodiment, the cancer is selected from the group consisting of ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, and non-Hodgkin cancer. Chitin lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, acute myeloid leukemia, multiple Myeloma, melanoma or mesothelioma.

Unless stated to the contrary, the following terms used in the specification and claims have the following meanings.

In the present invention, "C ₁ -C ₈ alkyl" refers to a straight-chain alkyl group and a branched alkyl group including 1 to 8 carbon atoms. The alkyl group refers to a saturated aliphatic hydrocarbon group, such as methyl, ethyl, n- Propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2 -Dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-tris Methylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl Methyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4 -Methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl , 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2- Dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl 2-ethylpentyl, 2-methyl-3-ethylpentyl, or various branched isomers thereof, and the like.

In the present invention, "cycloalkyl" refers to a saturated monocyclic hydrocarbon substituent, and "C ₃ -C ₈ alkylcycloalkyl" refers to a monocyclic cycloalkyl including 3 to 8 carbon atoms, for example, a monocyclic cycloalkyl Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

In the present invention, "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein one or more ring atoms are selected from nitrogen, oxygen, or S (O) r (where r is an integer of 0, Heteroatoms of 1, 2), but excluding the ring part of -OO-, -OS- or -SS-, the remaining ring atoms are carbon. "4-10 membered heterocyclyl" refers to a cyclic group containing 4 to 10 ring atoms. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups.

In the present invention, "alkenyl" refers to an alkyl group as defined above, which is composed of at least two carbon atoms and at least one carbon-carbon double bond, and "C ₂ -C ₈ alkenyl" refers to a straight chain containing 2 to 8 carbons. Or containing branched alkenyl. Examples are vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, and the like.

In the present invention, "alkynyl" refers to an alkyl group as defined above composed of at least two carbon atoms and at least one carbon-carbon triple bond, and "C ₂ -C ₈ alkenyl" refers to a straight-chain or Contains branched alkynyl. For example, ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl and the like.

In the present invention, "alkoxy" means -O- (alkyl), wherein alkyl is as defined above. "C ₃ -C ₈ alkoxy" refers to an alkyloxy group containing 1 to 8 carbons. Non-limiting examples include methoxy, ethoxy, propoxy, butoxy, and the like.

In the present invention, "cycloalkoxy" refers to -O- (unsubstituted cycloalkyl), wherein the definition of cycloalkyl is as described above. "C ₃ -C ₈ cycloalkoxy" refers to a cycloalkyloxy group containing 3-8 carbons. Non-limiting examples include cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy Wait.

"Halogen" means fluorine, chlorine, bromine or iodine.

"Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or a physiologically pharmaceutically acceptable salt or prodrug thereof with other chemical components, as well as other components such as physiologically pharmaceutically acceptable carriers and excipients.形 剂。 Shape agent. The purpose of the pharmaceutical composition is to promote the administration to the organism, which is beneficial to the absorption of the active ingredient and then exerts the biological activity.

In the preparation steps of the present invention, the abbreviations of the reagents used represent:

THF: Tetrahydrofuran

EA, ethyl acetate

PE, petroleum ether

DCM: Dichloromethane

MTBE, methyl tert-butyl ether

TFAA Trifluoroacetic anhydride

Pd ₂ (dba) ₃ tris (dibenzylideneacetone) dipalladium

Xantphos 4,5-bisdiphenylphosphine-9,9-dimethylxanthene

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 Nuclear magnetic resonance proton spectrum of the compound of Example 1.

Figure 2 Mass spectrum of the compound of Example 1.

Fig. 3 Nuclear magnetic resonance proton spectrum of the compound of Example 2;

Figure 4 Mass spectrum of the compound of Example 2.

Fig. 5 Nuclear magnetic resonance proton spectrum of the compound of Example 3;

Figure 6 Mass spectrum of the compound of Example 3.

Fig. 7 Nuclear magnetic resonance proton spectrum of the compound of Example 4;

Figure 8 Mass spectrum of the compound of Example 4.

Fig. 9 Nuclear magnetic resonance proton spectrum of the compound of Example 5;

Figure 10 Mass spectrum of the compound of Example 5.

Fig. 11 Nuclear magnetic resonance proton spectrum of the compound of Example 6;

Figure 12 Mass spectrum of the compound of Example 6.

Fig. 13 Nuclear magnetic resonance proton spectrum of the compound of Example 7;

Figure 14 Mass spectrum of the compound of Example 7.

Fig. 15 Nuclear magnetic resonance proton spectrum of the compound of Example 8;

Figure 16 Mass spectrum of the compound of Example 8.

Fig. 17 Nuclear magnetic resonance proton spectrum of the compound of Example 9;

Figure 18 Mass spectrum of the compound of Example 9.

Figure 19 Nuclear magnetic resonance proton spectrum of the compound of Example 10.

Figure 20 Mass spectrum of the compound of Example 10.

Fig. 21 Nuclear magnetic resonance proton spectrum of the compound of Example 11;

Figure 22 Mass spectrum of the compound of Example 11.

Fig. 23 Nuclear magnetic resonance proton spectrum of the compound of Example 12;

Figure 24 Mass spectrum of the compound of Example 12.

Fig. 25 Nuclear magnetic resonance proton spectrum of the compound of Example 13;

Fig. 26 Nuclear magnetic resonance proton spectrum of the compound of Example 14;

Figure 27 Mass spectrum of the compound of Example 14.

Figure 28 Nuclear magnetic resonance proton spectrum of the compound of Example 15

Figure 29 Mass spectrum of the compound of Example 15.

Figure 30 Nuclear magnetic resonance proton spectrum of the compound of Example 16.

Figure 31 Mass spectrum of the compound of Example 16.

Figure 32 Mass spectrum of the compound of Example 17.

Fig. 33 Nuclear magnetic resonance proton spectrum of the compound of Example 18;

Figure 34 Mass spectrum of the compound of Example 18.

detailed description

The present invention is described below with reference to specific examples. Those skilled in the art can understand that these examples are only used to illustrate the present invention, and they do not limit the scope of the present invention in any way.

Unless otherwise specified, the experimental methods in the following examples are conventional methods. Unless otherwise specified, the medicinal materials, reagent materials, etc. used in the following examples are all commercially available products.

Example 1

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) aminopyrimidin-2-yl) amino) -2-((2- (dimethyl Synthesis of Amino) ethyl) (Meth) amino) -6-methoxypyridin-3-yl) acrylamide

Step 1: Synthesis of 6-bromo-2-methoxy-3-nitropyridine

At room temperature, 2,6-dibromo-3-nitropyridine (40.00g, 141.90mmol) and THF (520ml) were added to the four-necked flask in order, the temperature was lowered to 0-5 ° C, and sodium methoxide (30%, 28.11g) , 156.08 mmol), 3h reaction stopped. The reaction solution was poured into ice water (500 ml), and extracted with MTBE (500 ml x 3). The organic phases were combined, washed with saturated brine (200 ml), the organic phase was concentrated, and the crude product was crystallized to obtain 19.89 g of a pale yellow solid with a yield of 60%. . ¹ H NMR (400 MHz, Chloroform-d) δ 8.15 (d, J = 8.2 Hz, 1 H), 7.22 (d, J = 8.2 Hz, 1 H), 4.14 (s, 3 H).

Step 2: Synthesis of 6-bromo-2-methoxypyridine-3-amine

At room temperature, 6-bromo-2-methoxy-3-nitropyridine (9.50 g, 38.62 mmol), iron powder (11.50 g, 193.11 mmol), and ammonium chloride (10.80 g, 193.11 mmol), ethanol (190 ml), water (95 ml), nitrogen protection, heating to 80 ° C., the reaction was terminated in 1 h. The reaction solution was filtered with diatomaceous earth, and the filtrate was concentrated. Water (950 ml) and EA (380 ml x 2) were added for extraction. The organic phases were combined, washed with saturated brine (200 ml), and concentrated to obtain 8.28 g of a black oily liquid with a yield of 100%. ¹ H NMR (400 MHz, Chloroform-d) δ 6.86 (d, J = 7.8 Hz, 1 H), 6.76 (d, J = 7.8 Hz, 1 H), 3.98 (s, 3 H).

Step 3: Synthesis of N- (6-bromo-2-methoxypyridin-3-yl) acetamide

At room temperature, 6-bromo-2-methoxypyridin-3-amine (12.00 g, 59.10 mmol), acetic anhydride (48.67 g, 466.90 mmol) were sequentially added to the four-necked flask, and the reaction was terminated at 20 ° C for 1 h. The reaction solution was added with water (240 ml) and stirred for 1 h, a brown precipitate was precipitated, filtered with suction, and the filter cake was dried to obtain 9.80 g of a brown solid with a yield of 68%. ¹ H NMR (400 MHz, Chloroform-d) δ 8.49 (d, J = 8.2 Hz, 1H), 7.06 (d, J = 8.2 Hz, 1H), 4.02 (s, 3H), 2.21 (s, 3H).

Step 4: Synthesis of N- (6-bromo-2-methoxy-5-nitro-pyridin-3-yl) acetamide

At room temperature, N- (6-bromo-2-methoxypyridin-3-yl) acetamide (6.80 g, 27.74 mmol) and trifluoroacetic anhydride (68 ml) were sequentially added to the four-necked flask, and the temperature was lowered to 0 ° C. Hereinafter, fuming nitric acid (2.00 g, 28.58 mmol) was added dropwise, and the reaction was terminated for 0.5 h. The reaction solution was slowly poured into ice water (500 g) and stirred for 1 h. An off-white precipitate was precipitated, filtered with suction, and the filter cake was dried to obtain 7.60 g of a yellow solid with a yield of 94%. ¹ H NMR (400 MHz, DMSO-d ₆ ) 9.12 (s, 1H), 4.06 (s, 3H), 2.16 (s, 3H).

Step 5: Synthesis of N- (6-((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxy-5-nitro-pyridin-3-yl) acetamide

At room temperature, N- (6-bromo-2-methoxy-5-nitro-pyridin-3-yl) acetamide (5.60 g, 19.30 mmol), N, N, N ' -Trimethylethylenediamine (2.96g, 28.96mmol), acetonitrile (168ml), warmed to 80 ° C, and the reaction was terminated in 1h. The solvent was distilled off under reduced pressure at 50 ° C, EA (70 ml) was slurried, suction filtered, washed with EA (20 ml), and the filter cake was dried to obtain 5.40 g of a yellow powder with a yield of 72%. ¹ H NMR (400MHz, DMSO-d ₆ ) 8.74 (s, 1H), 4.03 (s, 3H), 3.92 (br, 2H), 3.41 (t, J = 6.7Hz, 2H), 3.32 (s, 3H) , 2.84 (d, J = 4.6 Hz, 6H), 2.07 (s, 3H).

Step 6: N ² - (2- (dimethylamino) ethyl) ^-6-methoxy-2 -N - Synthesis of methyl 3-nitropyridine-2,5-diamine

At room temperature, N- (6-((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxy-5-nitro-pyridine-3- Alkyl) acetamide (3.60 g, 11.34 mmol), methanol (90 ml), 37% hydrochloric acid (2.22 g, 22.68 mmol), the temperature was raised to 60 ° C, and the reaction was terminated in 3 h. The reaction solution was neutralized by adding 5% NaHCO ₃ to pH = 8-9, and the solvent was distilled off under reduced pressure at 40 ° C. Water (30 ml) was added, followed by extraction with DCM (100 ml x 3). The organic phases were combined, concentrated, and subjected to crude column chromatography. Brownish yellow oil, 0.70 g, yield 22%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.46 (s, 1H), 4.73 (br, 2H), 3.93 (s, 3H), 3.59-3.54 (m, 2H), 2.77 (s, 3H), 2.52 (s, 2H), 2.17 (s, 6H).

Step 7: Synthesis of 2- (isopropylthio) -3-nitropyridine

At room temperature, add 5-fluoro-3-nitropyridine (176g, 1.24mol), potassium bicarbonate (342g, 2.47mol), DMF (2600ml), isopropyl mercaptan (101g, 1.36) to a 5L four-necked bottle in this order. mol), kept at 25 ° C., and the reaction was terminated for 3 h. The reaction solution was added with water (1000 ml) and extracted with PE (1000 ml x 4). The organic phases were combined, washed with water (500 ml) and washed once. The organic phase was concentrated to obtain 220.0 g of a red-brown liquid with a yield of 89%. ¹ H NMR (400MHz, Chloroform-d) δ 8.67 (dd, J = 4.6, 1.6 Hz, 1H), 8.45 (dd, J = 8.2, 1.6 Hz, 1H), 7.16 (dd, J = 8.2, 4.6 Hz , 1H), 4.15 (hept, J = 6.9Hz, 1H), 1.42 (d, J = 6.9Hz, 6H).

Step 8: Synthesis of 2- (isopropylsulfonyl) -3-nitropyridine

At room temperature, a 5-L four-necked flask was sequentially charged with 2- (isopropylthio) -3-nitropyridine (175.5 g, 885.29 mmol), 85% m-chloroperoxybenzoic acid (449 g, 2.21 mol), and DCM. (3500ml), the temperature was raised to 25-30 ° C, and the reaction was terminated in 1h. Pour the reaction solution into water (900ml), quench by adding 10% sodium sulfite solution (2000ml), adjust the pH = 8-9 by adding 20% potassium carbonate, separate the lower organic phase, add DCM (1500mlx2) for extraction, and combine the organics Phase, add 5% NaHCO ₃ (1000ml) and wash once. The organic phase is concentrated. The crude product is added with n-hexane (1000ml) and slurried for 0.5h, filtered with suction, and the filter cake is dried to obtain 155g of pale yellow solid with a yield of 76%. ¹ H NMR (400MHz, Chloroform-d) δ 8.91 (dd, J = 4.7, 1.3 Hz, 1H), 8.13 (dd, J = 8.1, 1.3 Hz, 1H), 7.76 (J = 8.1, 4.7 Hz, 1H ), 4.05 (hept, J = 6.9Hz, 1H), 1.42 (d, J = 6.9Hz, 6H).

Step 9: Synthesis of 2- (isopropylsulfonyl) pyridin-3-amine

At room temperature, 2- (isopropylsulfonyl) -3-nitropyridine (155 g, 673.21 mmol), iron powder (188 g, 3.37 mol), and ammonium chloride (180 g, 3.37 mol) were sequentially added to the four-necked flask. And THF (1550ml), water (1550ml), nitrogen protection, warmed to 70 ° C, the reaction was terminated in 6h. The reaction solution was filtered with diatomaceous earth, and the filtrate was concentrated. The water was added (1500 ml) and extracted with DCM (500 ml x 3). The organic phases were combined, washed with water (500 ml) and washed once. The organic phase was concentrated to obtain 140 g of crude product and crystals. The yield is 77%. ¹ H NMR (400MHz, Chloroform-d) δ 8.05 (d, J = 4.0 Hz, 1H), 7.26-7.22 (m, 1H), 7.05 (d, J = 8.4 Hz, 1H), 5.29 (br, 2H ), 3.74 (dq, J = 13.8, 7.0 Hz, 1H), 1.35 (d, J = 7.0 Hz, 3H).

Step 10: Synthesis of 2,5-dichloro-N- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidin-4-amine

At room temperature, 2- (isopropylsulfonyl) pyridine-3-amine (18.00 g, 89.88 mmol), 2,4,5-trichloropyrimidine (25.00 g, 134.82 mmol), and DMF were sequentially added to the four-necked flask. (180ml), protected by nitrogen, lowered the temperature to 15-20 ° C, and added 60% sodium hydrogen (10.80g, 269.64mmol) several times in a small amount, kept at 20 ° C, and the reaction was terminated in 1h. The reaction solution was slowly poured into a solution of acetic acid (8g) in ice-water (720ml) to quench, precipitate was precipitated, and isopropyl ether (360ml) was added to beat and disperse for 0.5h, suction-filtered, and the filter cake was dried to obtain 27.00g of a yellowish solid The yield was 86.4%. ¹ H NMR (400 MHz, Chloroform-d) 9.23 (d, J = 8.7 Hz, 1 H), 8.43 (d, J = 4.3 Hz, 1 H), 8.32 (d, J = 1.3 Hz, 1 H), 7.61 (dd, J = 8.1, 3.8 Hz, 1H), 3.94 (hept, J = 8.3, 7.5 Hz, 1H), 1.39-1.36 (m, 6H).

Step 11: 5-chloro-N ^2- (6-((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxy-5-nitro-pyridin-3-yl) Synthesis of -N ^4- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidine-2,4-diamine

At room temperature was added successively a four-necked flask N ² - (2- (dimethylamino) ethyl) -6-methoxy -N ² - methyl-3-nitropyridine-2,5-diamine (0.70 g, 2.60 mmol), 2,5-dichloro-N- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidin-4-amine) (0.90 g, 2.60 mmol), Pd ₂ ( dba) ₃ (0.035 g, 0.039 mmol), Xantphos (0.045 g, 0.078 mmol), K ₃ PO ₄ (1.380 g, 6.50 mmol), and 1,4-dioxane (21 ml), protected by nitrogen, and heated to 80 At 3 ° C, the reaction was terminated. The reaction solution was cooled to 25-35 ° C and quenched by adding water (40 ml), and extracted by adding EA (20 ml x 2). The organic phases were combined, washed with saturated brine (20 ml), and the organic phase was concentrated. Brown-yellow solid 0.24g, yield 16%. MS (ESI) m / z: 580.1 [M + H] ⁺ .

Step 12: N ⁵ - (5- chloro-4 - ((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) -N ² - (2- (dimethylamino ) Ethyl) -6-methoxy-N ² - methylpyridine 2,3,5-triamine

At room temperature, 5-chloro-N ^2- (6-((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxy-5-nitro -Pyridin-3-yl) -N ^4- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidine-2,4-diamine (0.225 g, 0.39 mmol), iron powder (0.109 g, 1.94 mmol), ammonium chloride (0.104 g, 1.94 mmol) and ethanol (9 ml), water (4.5 ml), protected by nitrogen, heated to 80 ° C., and the reaction was terminated in 1 h. The reaction solution was filtered by adding diatomaceous earth, and the filtrate was concentrated. The crude product was subjected to column chromatography to obtain 0.16 g of a brown-yellow solid with a yield of 75%.

Step 13: N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2-((2- Synthesis of (dimethylamino) ethyl) (meth) amino) -6-methoxypyridin-3-yl) acrylamide

At room temperature, 4-neck flask were added to N ⁵ - (5- chloro-4 - ((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) -N ² - ( 2- (dimethylamino) ethyl) -6-methoxy-N ² - methylpyridine 2,3,5-triamine (1.60 g, 2.91 mmol), dichloromethane (32 ml), nitrogen protection, The temperature was lowered to below -30 ° C, and a solution of acrylic anhydride (0.455 g, 4.36 mmol) in DCM (1 ml) was added dropwise, and the reaction was terminated in 4 h. The reaction solution was quenched by adding 5% NaHCO ₃ (100 ml), and extracted by adding EA (50 ml x 5). The organic phases were combined, concentrated, and subjected to crude column chromatography to obtain a brown-yellow solid 0.14 g with a yield of 10%. ¹ H NMR (400MHz, DMSO-d ₆ ) 8.69 (s, 1H), 8.37 (s, 1H), 8.24 (s, 1H), 8.06 (s, 1H), 7.51 (s, 1H), 6.53-6.37 ( m, 1H), 6.18 (d, J = 17.4 Hz, 1H), 5.73 (d, J = 11.3 Hz, 1H), 3.87 (dd, J = 13.6, 6.8 Hz, 1H), 3.82 (s, 3H), 3.43 (br, 2H), 3.22 (br, 2H), 2.87 (s, 3H), 2.22 (s, 6H), 1.22 (d, J = 6.8 Hz, 6H). MS (ESI) m / z: 604.1 [M + H] ⁺ .

Example 2

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethylform Of phenylamino) ethyl) (meth) amino) -6-ethoxypyridin-3-yl) acrylamide

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethylform The method for preparing aminoamino) ethyl) (meth) amino) -6-ethoxypyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) 8.61 (s, 1H), 8.36 (d, J = 4.0Hz, 1H), 8.24 (s, 1H), 8.06 (s, 1H), 7.48 (s, 1H) , 6.43 (dd, J = 17.0, 10.2 Hz, 1H), 6.21-6.14 (m, 1H), 5.75-5.68 (m, 1H), 4.29 (q, J = 7.0 Hz, 2H), 3.86 (dt, J = 13.5, 6.8Hz, 1H), 3.18 (t, J = 6.3Hz, 2H), 2.85 (s, 3H), 2.46 (d, J = 6.7Hz, 2H), 2.19 (s, 6H), 1.23 (s , 9H). MS (ESI) m / z: 618.1 [M + H] ⁺ .

Example 3

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethylform Of phenylamino) ethyl) (meth) amino) -6-isopropoxypyridin-3-yl) acrylamide

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethylform The method for preparing aminoamino) ethyl) (meth) amino) -6-isopropoxypyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) 9.87 (br, 1H), 9.75 (br, 1H), 8.98 (br, 1H), 8.52 (s, 1H), 8.36 (d, J = 3.8 Hz, 1H) , 8.25 (s, 1H), 8.05 (s, 1H), 7.47 (s, 1H), 6.43 (dd, J = 17.0, 10.2Hz, 1H), 6.21-6.13 (m, 1H), 5.76-5.69 (m , 1H), 5.14 (p, J = 6.1 Hz, 1H), 3.85 (dt, J = 13.8, 6.9 Hz, 1H), 3.17 (t, J = 6.4 Hz, 2H), 2.84 (s, 3H), 2.46 -2.42 (m, 2H), 2.18 (s, 6H), 1.21 (dd, J = 10.5, 6.5Hz, 12H). MS (ESI) m / z: 632.1 [M + H] ⁺ .

Example 4

N- (5-((5-chloro-4- (2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethyl Synthesis of Amino) ethyl) (Meth) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide

N- (5-((5-chloro-4- (2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethyl Amino) ethyl) (meth) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide is prepared in a similar manner to Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.91 (br, 1H), 9.84 (br, 1H), 8.96 (br, 1H), 8.83 (s, 1H), 8.37 (s, 1H), 8.24 ( s, 1H), 7.99 (s, 1H), 7.45 (s, 1H), 6.45 (dd, J = 16.9, 10.2 Hz, 1H), 6.20 (d, J = 16.8 Hz, 1H), 5.75 (d, J = 10.4Hz, 1H), 4.89 (q, J = 8.9Hz, 2H), 3.85 (dt, J = 13.4, 6.6Hz, 1H), 3.28-3.22 (m, 2H), 2.89 (s, 3H), 2.46 -2.42 (m, 2H), 2.20 (s, 6H), 1.22 (d, J = 6.7Hz, 6H). MS (ESI) m / z: 672.1 [M + H] ⁺ .

Example 5

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6- (difluoromethoxy) Synthesis of 2-((2- (dimethylamino) ethyl) (meth) amino) pyridin-3-yl) acrylamide

Step 1: Synthesis of 2-chloro-6- (difluoromethoxy) pyridine

At room temperature, 6-chloro-2-hydroxypyridine (148.8 g, 1.15 mol), DMF (1500 ml), K ₂ CO ₃ (317 g, 1.15 mol) were sequentially added to the four-necked flask, and sodium difluorochloroacetate ( 210 g, 1.38 mol), the temperature was raised to 85 ° C. and the temperature was maintained for 1 h, and then the temperature was maintained at 100 ° C. for 4 h, and the reaction was terminated. The solvent was distilled off under reduced pressure at 30 ° C, water (20 ml) was added, and EA (20 ml) was added twice for extraction. The organic phases were combined, washed with water (10 ml), and the organic phase was concentrated. The crude column chromatography yielded 133.60 g of a pale yellow solid. The yield is 65%. ¹ H NMR (400 MHz, Chloroform-d) δ 7.71 (t, J = 7.9 Hz, 1 H), 7.55 (t, J = 72.5 Hz, 1 H), 7.16 (d, J = 7.7 Hz, 1 H), 6.85 ( d, J = 8.1 Hz, 1H).

Step 2: Synthesis of 6-chloro-2- (difluoromethoxy) -3-nitropyridine

At room temperature, 2-chloro-6- (difluoromethoxy) pyridine (40.00g, 222.78mmol), and trifluoroacetic anhydride (120ml) were added to the four-necked flask in this order. Fuming nitric acid (16.00 g, 229.46 mmol), and react at 0-10 ° C for 2h. Ice water (1000 ml) was added, and methyl tertiary ether (300 ml x 3) was extracted. The organic phases were combined, concentrated, and column chromatography gave 25.00 g of a pale yellow solid with a yield of 50%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.67 (d, J = 8.4 Hz, 1 H), 7.76 (t, J = 70.5 Hz, 1 H), 7.66 (d, J = 8.4 Hz, 1 H).

Step 3: Synthesis of 6-chloro-2- (difluoromethoxy) pyridin-3-amine

At room temperature, 6-chloro-2- (difluoromethoxy) -3-nitropyridine (43.70 g, 194.61 mmol), iron powder (54.30 g, 973.06 mmol), and ammonium chloride were sequentially added to the four-necked flask. (52.00g, 973.06mmol) and ethanol (875ml), water (437ml), nitrogen protection, warmed to 80 ° C, the reaction was terminated in 2h. The reaction solution was filtered by adding diatomaceous earth, and the filtrate was concentrated. The crude product was subjected to column chromatography to obtain 34.20 g of a brown liquid with a yield of 90%. ¹ H NMR (400 MHz, Chloroform-d) δ 7.44 (t, J = 72.7 Hz, 1 H), 7.02 (d, J = 8.1 Hz, 1 H), 6.95 (d, J = 8.1 Hz, 1 H).

Step 4: Synthesis of N- (6-chloro-2- (difluoromethoxy) pyridin-3-yl) acetamide

At room temperature, 6-chloro-2- (difluoromethoxy) pyridine-3-amine (34.20 g, 175.77 mmol), acetic anhydride (171 g, 1.67 mol) were sequentially added to the four-necked flask, and reacted at 20-25 ° C. , 1h reaction was terminated. The reaction solution was added with water (800 ml) and stirred for 1 h. A brown precipitate was precipitated, extracted with DCM (200 ml x 3), washed once with 100 ml of water, and the combined organic phases were concentrated to give 28.60 g of a yellow solid with a yield of 69%.

Step 5: Synthesis of N- (6-chloro-2- (difluoromethoxy) -5-nitro-pyridin-3-yl) acetamide

At room temperature, N- (6-chloro-2- (difluoromethoxy) pyridin-3-yl) acetamide (22.60 g, 95.52 mmol), and trifluoroacetic anhydride (113 ml) were sequentially added to the four-necked flask. Add fuming nitric acid (30.1 g, 477.60 mmol) dropwise and react at 70 ° C for 16h. The reaction solution was slowly poured into ice-water (1000 ml) and stirred for 1 h, extracted with methyl tert-ether (300 ml x 3), the organic phases were combined, concentrated, and column chromatography gave 3.40 g of a brown solid with a yield of 13%. ¹ H NMR (400 MHz, Chloroform-d) δ 9.44 (s, 1 H), 7.70 (d, J = 5.6 Hz, 1 H), 7.53 (t, J = 72.5 Hz, 1 H), 2.33 (s, 3 H).

Step 6: N- (2- (difluoromethoxy) -6-((2- (dimethylamino) ethyl) (methyl) amino) -5-nitro-pyridin-3-yl) ethyl Synthesis of amides

At room temperature, N- (6-chloro-2- (difluoromethoxy) -5-nitro-pyridin-3-yl) acetamide (1.70 g, 6.04 mmol), N, N, N'-trimethylethylenediamine (1.23g, 12.08mmol), acetonitrile (34ml), the temperature was raised to 80 ° C, and the reaction was terminated in 3h. The solvent was evaporated under reduced pressure at 50 ° C, 5% NaHCO ₃ (20 ml) was added, and extracted with DCM (30 ml x 3) three times. The organic phases were combined, concentrated, and subjected to crude column chromatography to obtain 1.01 g of a brown solid with a yield of 48. %. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.61 (s, 1H), 8.64 (s, 1H), 7.76 (t, J = 71.8Hz, 1H), 3.65 (t, J = 6.9Hz, 2H) , 2.83 (s, 3H), 2.53 (d, J = 8.0 Hz, 2H), 2.19 (s, 6H), 2.07 (s, 3H).

Step 7: 6- (difluoromethoxy) -N ² - (2- (dimethylamino) ethyl) ² -N - Synthesis of methyl 3-nitropyridine-2,5-diamine

At room temperature, N- (2- (difluoromethoxy) -6-((2- (dimethylamino) ethyl) (methyl) amino) -5-nitro- Pyridine-3-yl) acetamide (1.00 g, 2.88 mmol), methanol (30 ml), 37% hydrochloric acid (0.56 g, 5.76 mmol), the temperature was raised to 60 ° C, and the reaction was terminated in 6 h. The reaction solution was neutralized with 5% NaHCO ₃ to Ph = 8-9, the solvent was distilled off under reduced pressure at 40 ° C, water (6 ml) was added, and DCM (30 ml x 3) was added for extraction. The organic phases were combined, concentrated, and subjected to crude column chromatography to obtain 0.5 g of brown-yellow oil, 57% yield, directly into the next step.

Step 8: 5-Chloro -N ² - (2- (difluoromethoxy) -6 - ((2- (dimethylamino) ethyl) (methyl) amino) -5-nitro-pyridin-3 -Yl) -N ^4- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidine-2,4-diamine

At room temperature, a four-necked flask were added 6- (difluoromethoxy) -N ² - (2- (dimethylamino) ethyl) -N ² - methyl-3-nitro-2, Synthesis of 5-diamine (0.50g, 1.00eq), 2,5-dichloro-N- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidin-4-amine (0.57g, 1.64mmol ), Pd ₂ (dba) ₃ (0.15 g, 0.16 mmol), Xantphos (0.19 g, 0.32 mmol), K ₃ PO ₄ (0.87 g, 4.09 mmol), and 1,4-dioxane (15 ml), nitrogen Protected, warmed to 60-70 ° C, the reaction was terminated in 3h. The reaction solution was cooled to 25-35 ° C, water (20 ml) was added to quench the reaction, and DCM (20 ml x 2) was added for extraction. The organic phases were combined, washed with saturated brine (10 ml), and the organic phase was concentrated. The crude column chromatography yielded Brownish yellow solid 0.25g, yield 25%, directly into the next step.

Step 9: N ⁵ - (5- chloro-4- (2- (isopropylsulfonyl) pyridin-3-ylamino) pyrimidin-2-yl) -6- (difluoromethoxy) -N ² - Synthesis of (2- (dimethylamino) ethyl) -N2-methylpyridin-2-yl) 2,3,5-triamine

At room temperature, 4-neck flask were added to the 5-Chloro -N ² - (2- (difluoromethoxy) -6 - ((2- (dimethylamino) ethyl) (methyl) amino) - 5-nitropyridin-3-yl) -N ^4- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidine-2,4-diamine (0.25 g, 4.06 mmol), iron powder (0.11 g, 20.30 mmol), ammonium chloride (0.11 g, 20.30 mmol), ethanol (5 ml), water (2.5 ml), nitrogen protection, heating to 80 ° C., and the reaction was terminated in 2 h. The reaction solution was filtered by adding diatomaceous earth, and the filtrate was concentrated. The crude product was subjected to column chromatography to obtain 0.09 g of a yellow-green solid with a yield of 38%.

Step 10: N- (5- (5-chloro-4- (2- (isopropylsulfonyl) pyridin-3-ylamino) pyrimidin-2-ylamino) -6- (difluoromethoxy)- Synthesis of 2-((2- (dimethylamino) ethyl) (meth) amino) pyridin-3-yl) acrylamide

At room temperature was added successively a four-necked flask N ⁵ - (5-chloro-4- (2- (isopropylsulfonyl) pyridin-3-ylamino) pyrimidin-2-yl) -6- (difluoromethyl oxy) -N ² - (2- (dimethylamino) ethyl) -N2- methyl-pyridin-2-yl) 2,3,5-triamine (0.09g, 0.15mmol), dichloromethane ( 4.5ml), protected by nitrogen, cooled to below -30 ° C, a solution of acrylic anhydride (0.014g, 0.15mmol) in DCM (1ml) was added dropwise, and the reaction was terminated in 1h. The reaction solution was quenched by adding 5% NaHCO ₃ (15 ml), and extracted by adding DCM (10 ml x 2). The organic phases were combined, concentrated, and subjected to crude column chromatography. A scrape was obtained to obtain 0.052 g of an off-white solid with a yield of 52%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.08 (br, 1H), 9.91 (s, 1H), 9.03 (s, 1H), 8.38 (d, J = 4.1 Hz, 1H), 8.26 (s, 1H), 8.14 (s, 1H), 7.63 (t, J = 71.8Hz, 1H), 7.54 (s, 1H), 6.61-6.55 (m, 1H), 6.21 (d, J = 11Hz, 1H), 5.78-5.73 (m, 1H), 3.88-3.83 (m, 1H), 3.43-3.37 (m, 2H), 2.87 (s, 3H), 2.49-2.44 (m, 2H), 2.28 (s, 6H), 1.22 (d, J = 6.9Hz, 6H). MS (ESI) m / z: 640.1 [M + H] ⁺ .

Example 6

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) phenyl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethylamino) Synthesis of (ethyl) (meth) amino) -6-methoxypyridin-3-yl) acrylamide

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) phenyl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethylamino) The method for preparing ethyl) (meth) amino) -6-methoxypyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) 8.57 (s, 1H), 8.50 (br, 1H), 8.21 (s, 1H), 8.08 (s, 1H), 7.77 (d, J = 7.6Hz, 1H) , 7.53 (s, 1H), 7.26 (t, J = 7.6 Hz, 1H), 6.43 (dd, J = 16.9, 10.2 Hz, 1H), 6.17 (d, J = 16.8 Hz, 1H), 5.72 (d, J = 10.1 Hz, 1H), 3.81 (s, 3H), 3.43 (dd, J = 10.1, 3.6 Hz, 1H), 3.20-3.16 (m, 2H), 2.86 (s, 3H), 2.44 (d, J = 6.8Hz, 2H), 2.18 (s, 6H), 1.16 (d, J = 6.7Hz, 6H). MS (ESI) m / z: 603.1 [M + H] ⁺ .

Example 7

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((5-fluoro-4-((2- (isopropylsulfonyl) pyridine-3- Synthesis of amino) amino) pyrimidin-2-yl) amino) -6-methoxypyridin-3-yl) acrylamide

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((5-fluoro-4-((2- (isopropylsulfonyl))

The method for preparing pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6-methoxypyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) 8.44 (s, 1H), 8.38 (d, J = 4.2Hz, 1H), 8.21 (d, J = 3.0Hz, 1H), 8.17 (s, 1H), 7.55 (dd, J = 8.5, 4.2 Hz, 1H), 6.44 (dd, J = 17.0, 10.2 Hz, 1H), 6.18 (dd, J = 17.0, 1.8 Hz, 1H), 5.77-5.68 (m, 1H), 3.89 (dd, J = 13.6, 6.8Hz, 1H), 3.84 (s, 3H), 3.18 (t, J = 6.3Hz, 2H), 2.85 (s, 3H), 2.48-2.43 (m, 2H), 2.19 (s, 6H), 1.22 (d, J = 6.8 Hz, 6H).

MS (ESI) m / z: 588.1 [M + H] ⁺ .

Example 8

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((5-fluoro-4-((2- (isopropylsulfonyl) pyridine-3- Synthesis of amino) amino) pyrimidin-2-yl) amino) -6-isopropoxypyridin-3-yl) acrylamide

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((5-fluoro-4-((2- (isopropylsulfonyl) pyridine-3- The method for preparing the group) amino) pyrimidin-2-yl) amino) -6-isopropoxypyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, Chloroform-d) δ 10.09 (br, 1H), 9.93 (br, 1H), 9.44 (br, 1H), 9.15 (s, 1H), 8.32 (s, 1H), 8.14 (s , 1H), 7.40 (dd, J = 8.8, 4.2Hz, 1H), 7.19 (s, 1H), 6.44-6.19 (m, 2H), 5.70 (dd, J = 8.1, 3.6Hz, 1H), 5.39- 5.23 (m, 1H), 3.91-3.76 (m, 1H), 2.94 (s, 2H), 2.73 (s, 3H), 2.40 (d, J = 5.9Hz, 2H), 2.34 (s, 6H), 1.39 (dd, J = 11.2, 6.6 Hz, 12H); MS (ESI) m / z: 616.1 [M + H] ⁺ ; HPLC = 99.84%. .

Example 9

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((5-fluoro-4-((2- (isopropylsulfonyl) pyridine-3- (Synthetic) amino) pyrimidin-2-yl) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((5-fluoro-4-((2- (isopropylsulfonyl) pyridine-3- (Meth) amino) pyrimidin-2-yl) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide is prepared in a similar manner to Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) 9.83 (br, 2H), 9.00 (br, 1H), 8.59 (s, 1H), 8.38 (s, 1H), 8.21 (d, J = 3.0Hz, 1H) , 8.07 (s, 1H), 7.50 (dd, J = 8.7, 4.4 Hz, 1H), 6.44 (dd, J = 17.0, 10.2 Hz, 1H), 6.20 (dd, J = 17.0, 1.8 Hz, 1H), 5.78-5.72 (m, 1H), 4.91 (q, J = 9.1Hz, 2H), 3.88-3.84 (m, 1H), 3.22 (t, J = 6.4Hz, 2H), 2.88 (s, 3H), 2.47 -2.44 (m, 2H), 2.18 (s, 6H), 1.21 (d, J = 6.7Hz, 6H). MS (ESI) m / z: 656.1 [M + H] ⁺ .

Example 10

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) Synthesis of -5-methylpyrimidin-2-yl) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) The method for preparing 5-methylpyrimidin-2-yl) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.84 (s, 1H), 9.42 (s, 1H), 9.07 (d, J = 8.6 Hz, 1H), 8.30 (d, J = 3.8 Hz, 1H) , 8.28 (s, 1H), 8.17 (s, 1H), 8.00 (s, 1H), 7.45 (dd, J = 8.5, 4.3Hz, 1H), 6.49-6.40 (m, 1H), 6.19 (d, J = 16.8 Hz, 1H), 5.75 (d, J = 11.7 Hz, 1H), 4.96-4.89 (m, 2H), 3.84 (p, J = 6.8Hz, 1H), 3.26-3.19 (m, 2H), 2.86 (s, 3H), 2.47-2.44 (m, 2H), 2.23 (s, 6H), 2.08 (s, 3H), 1.22 (d, J = 6.8Hz, 6H). MS (ESI) m / z: 652.1 [M + H] ⁺ .

Example 11

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) Synthesis of pyrimidin-2-yl) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide

N- (2-((2- (dimethylamino) ethyl) (methyl) amino) -5-((4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) The method for preparing pyrimidin-2-yl) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) δ9.81 (s, 1H), 9.29 (s, 1H), 8.77 (br, 1H), 8.35 (s, 1H), 8.32 (d, J = 3.7Hz, 1H), 8.13 (s, 1H), 8.08 (d, J = 5.6Hz, 1H), 7.46 (s, 1H), 6.44 (dd, J = 17.0, 10.2Hz, 1H), 6.33 (d, J = 5.6 Hz, 1H), 6.19 (d, J = 17.0 Hz, 1H), 5.75 (d, J = 10.2 Hz, 1H), 4.92 (q, J = 9.1 Hz, 2H), 3.82 (dt, J = 13.6, 6.7 Hz, 1H), 3.19 (t, J = 5.9Hz, 2H), 2.86 (s, 3H), 2.48-2.43 (m, 2H), 2.19 (s, 6H), 1.20 (d, J = 6.8Hz, 6H MS (ESI) m / z: 638.1 [M + H] ⁺ .

Example 12

N- (5-((5-chloro-4-((2- (cyclopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethylform Of phenylamino) ethyl) (meth) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide

Step 1: Synthesis of 6- (N, N, N'-trimethylethylenediamino) -5-nitro-3-amino-2- (2,2,2-trifluoroethoxy) with reference to Example 4 ) Pyridine.

Step 2: Synthesis of sodium cyclopropane sulfonate

At room temperature, sodium sulfite (18.40 g, 145.54 mmol), sodium bicarbonate (20.70 g, 252.39 mmol), and water (180 ml) were sequentially added to the single-necked flask. The temperature was raised to 55-60 ° C, and cyclopropylsulfonyl chloride (18.60) was added dropwise. g, 132.31 mmol), the reaction was maintained at 60 ° C. for 3 h and the reaction was terminated. The reaction solution was concentrated, and THF (200 ml × 2) was added to bring it to constant weight to obtain 44.00 g of a white solid. The yield was calculated as 100%. The next step was directly fed without purification.

Step 3: Synthesis of 2- (cyclopropylsulfonyl) -3-nitropyridine

At room temperature, add 2-fluoro-3-nitropyridine (16.00 g, 112.60 mmol), sodium cyclopropane sulfonate (15.10 g, 118.23 mmol), and DMSO (160 ml) to the single-necked flask, and heat to 120 ° C for 4 h. The reaction was terminated. The reaction solution was cooled to 25-30 ° C, poured into water (480ml), precipitated out, filtered by suction, the filter cake was washed with water (100ml), and the filter cake was dissolved by adding DCM (200ml), the organic phase was separated, concentrated, and the column layer Analysis gave 16.50 g of off-white solid with a yield of 64%. ¹ H NMR (400 MHz, Chloroform-d) δ 8.90 (d, J = 4.7 Hz, 1 H), 8.18 (d, J = 8.1 Hz, 1 H), 7.74 (dd, J = 8.1, 4.7 Hz, 1 H), 3.13 (tt, J = 8.0, 4.8 Hz, 1H), 1.49-1.45 (m, 2H), 1.21-1.15 (m, 2H).

Step 4: Synthesis of 2- (cyclopropylsulfonyl) pyridine-3-amine

At room temperature, 2- (cyclopropylsulfonyl) -3-nitropyridine (15.80 g, 69.23 mmol), iron powder (19.30 g, 346.16 mmol), and ammonium chloride (18.50 g, 346.16 mmol), ethanol (316 ml), water (158 ml), nitrogen protection, heating to 80 ° C., the reaction was terminated in 4 h. The reaction solution was filtered with celite, the filtrate was concentrated, water (200 ml) was added, and DCM (100 ml × 2) was extracted. The organic phases were combined and concentrated to obtain 13.00 g of a brown-yellow solid with a yield of 95%.

Step 5: Synthesis of 2,5-dichloro-N- (2- (cyclopropylsulfonyl) pyridin-3-yl) pyrimidin-4-amine

At room temperature, 2- (cyclopropylsulfonyl) pyridin-3-amine (6.00 g, 30.26 mmol), 2,4,5-trichloropyrimidine (8.20 g, 45.39 mmol), and DMF were sequentially added to the four-necked flask. (60ml), protected by nitrogen, lowered the temperature to 0-5 ° C, and added 60% sodium hydrogen (2.40g, 60.52mmol) several times in small amounts, kept at 0 ° C, and the reaction was terminated in 3h. The reaction solution was slowly introduced into ice water (360 ml) for quenching, followed by extraction with DCM (100 ml × 2). The organic phases were combined, washed with water (100 ml), washed once, and the organic phase was concentrated. The crude product was added with isopropyl ether (60 ml) for 0.5 h. Filtration and drying of the filter cake gave 6.20 g of a khaki solid with a yield of 60%.

Step 6: 5-Chloro-N ^4- (2- (cyclopropylsulfonyl) pyridin-3-yl) -N ^2- (6-((2- (dimethylamino) ethyl) (methyl) Synthesis of Amino) -5-nitro-2- (2,2,2-trifluoroethoxy) pyridin-3-yl) pyrimidine-2,4-diamine

At room temperature, 6- (N, N, N'-trimethylethylenediamine) -5-nitro-3-amino-2- (2,2,2-trifluoroethyl) was sequentially added to the four-necked flask. Oxy) pyridine (6.60 g, 19.57 mmol), 2,5-dichloro-N- (2- (cyclopropylsulfonyl) pyridin-3-yl) pyrimidin-4-amine (6.80 g, 19.57 mmol), Pd ₂ (dba) ₃ (0.36g, 0.39mmol), Xantphos (0.45g, 8.81mmol), K ₃ PO ₄ (10.40g, 22.02mmol) and 1,4-dioxane (198ml), protected by nitrogen, The temperature was raised to 65 ° C, and the reaction was terminated in 11 hours. The reaction solution was cooled to 25-35 ° C and quenched by adding water (1000 ml), and extracted by adding DCM (300 ml x 2). The organic phases were combined, washed with water (200 ml) once, and the organic phase was concentrated. The crude column chromatography yielded a gray solid 7.00 g, yield 55%. MS (ESI) m / z: 646.1 [M + H] ⁺ .

Step 7: N ⁵ - (5- chloro-4 - ((2- (cyclopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) -N ² - (2- (dimethylamino ) Ethyl) -N ² -methyl-6- (2,2,2-trifluoroethoxy) pyridine-2,3,5-triamine

At room temperature, 5-chloro-N ^4- (2- (cyclopropylsulfonyl) pyridin-3-yl) -N ^2- (6-((2- (dimethylamino)) Ethyl) (methyl) amino) -5-nitro-2- (2,2,2-trifluoroethoxy) pyridin-3-yl) pyrimidine-2,4-diamine (4.00 g, 6.19 mmol ), Iron powder (1.73 g, 30.96 mmol), ammonium chloride (1.66 g, 30.96 mmol), ethanol (80 ml), water (40 ml), nitrogen protection, heating to 80 ° C., and the reaction was terminated in 1 h. The reaction solution was filtered with celite, the filtrate was concentrated, water (50 ml) was added, and DCM (30 ml x 2) was extracted. The organic phases were combined, concentrated, and the crude product was crystallized to obtain 1.85 g of a gray solid with a yield of 48%. MS (ESI) m / z: 616.1 [M + H] ⁺ .

Step 8: N- (5-((5-chloro-4-((2- (cyclopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2-((2- Synthesis of (dimethylamino) ethyl) (methyl) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide

At room temperature was added successively a four-necked flask N ⁵ - (5-chloro-4 - ((2- (cyclopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) -N ² - ( 2- (dimethylamino) ethyl) -N ² -methyl-6- (2,2,2-trifluoroethoxy) pyridine-2,3,5-triamine (1.00 g, 1.62 mmol) , Dichloromethane (20 ml), protected by nitrogen, cooled to below -30 ° C, a solution of acrylic anhydride (0.31 g, 2.43 mmol) in DCM (1 ml) was added dropwise, and the reaction was terminated in 1 h. The reaction solution was quenched by adding 5% NaHCO ₃ (30 ml), and extracted by adding DCM (20 ml x 2). The organic phases were combined, concentrated, and subjected to crude column chromatography to obtain 0.63 g of a brown solid, followed by DCM / isopropyl ether = 1.2 ml / 7.2 ml of crystals gave 0.31 g of off-white powder with a yield of 28%. ¹ H NMR (400MHz, DMSO-d ₆ ) 9.83 (br, 1H), 9.69 (br, 1H), 8.90 (br, 1H), 8.81 (s, 1H), 8.37 (s, 1H), 8.22 (s, 1H), 7.97 (s, 1H), 7.44 (s, 1H), 6.44 (dd, J = 16.9, 10.2 Hz, 1H), 6.21 (d, J = 18.5 Hz, 1H), 5.75 (d, J = 11.6 Hz, 1H), 4.90 (q, J = 9.1 Hz, 2H), 3.24 (t, J = 6.3 Hz, 2H), 3.18-3.16 (m, 1H), 2.89 (s, 3H), 2.46 (t, J = 5.8Hz, 2H), 2.19 (s, 6H), 1.19-1.10 (m, 4H). MS (ESI) m / z: 670.1 [M + H] ⁺ .

Example 13

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) phenyl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethylamino) Synthesis of (ethyl) (meth) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) phenyl) amino) pyrimidin-2-yl) amino) -2-((2- (dimethylamino) The method for preparing ethyl) (meth) amino) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.83 (s, 1H), 9.54 (s, 1H), 8.72 (s, 1H), 8.44 (br, 1H), 8.21 (s, 1H), 8.02 ( s, 1H), 7.77 (d, J = 7.7 Hz, 1H), 7.48 (s, 1H), 7.26 (t, J = 7.4 Hz, 1H), 6.45 (dd, J = 17.0, 10.0 Hz, 1H), 6.20 (d, J = 17.0 Hz, 1H), 5.74 (d, J = 10.0 Hz, 1H), 4.88 (q, J = 9.2 Hz, 2H), 3.44-3.38 (m, 1H), 3.24 (s, 2H ), 2.88 (s, 3H), 2.50-2.45 (m, 2H), 2.21 (s, 6H), 1.16 (d, J = 6.7Hz, 6H). MS (ESI) m / z: 671.1 [M + H] ⁺ .

Example 14

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6-methoxy-2- ( Synthesis of 4-methylpiperazin-1-yl) pyridin-3-yl) acrylamide

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6-methoxy-2- ( The method for preparing 4-methylpiperazin-1-yl) pyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, Chloroform-d) , 1H), 7.91 (s, 1H), 7.37 (s, 1H), 7.17 (s, 1H), 6.44-6.06 (m, 2H), 5.76 (d, J = 9.9Hz, 1H), 3.96 (s, 3H), 3.80 (p, J = 6.9Hz, 1H), 3.06-3.02 (m, 4H), 2.61-2.57 (m, 4H), 2.38 (s, 3H), 1.39 (d, J = 7.0Hz, 6H ). MS (ESI) m / z: 602.2 [M + H] ⁺ .

Example 15

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6-ethoxy-2- ( Synthesis of 4-methylpiperazin-1-yl) pyridin-3-yl) acrylamide

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6-ethoxy-2- ( The method for preparing 4-methylpiperazin-1-yl) pyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d6) δ 9.89 (br, 1H), 9.29 (br, 1H), 9.03 (br, 1H), 8.65 (s, 1H), 8.38 (d, J = 4.4 Hz, 1H ), 8.27 (s, 1H), 8.01 (s, 1H), 7.52 (s, 1H), 6.58 (dd, J = 17.0, 10.2 Hz, 1H), 6.18 (dd, J = 17.1, 2.1 Hz, 1H) , 5.73 (dd, J = 10.2, 2.1 Hz, 1H), 4.31 (q, J = 7.1 Hz, 2H), 3.88 (p, J = 6.8 Hz, 1H), 3.25-3.04 (m, 4H), 2.49 ( d, J = 5.2Hz, 4H), 2.24 (s, 3H), 1.25-1.23 (m, 9H). MS (ESI) m / z: 616.1 [M + H] ⁺ .

Example 16

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6-isopropyl-2- ( Synthesis of 4-methylpiperazin-1-yl) pyridin-3-yl) acrylamide

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6-isopropyl-2- ( The method for preparing 4-methylpiperazin-1-yl) pyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.86 (br, 1H), 9.25 (br, 1H), 8.94 (br, 1H), 8.55-8.48 (m, 1H), 8.35 (d, J = 3.7 Hz, 1H), 8.25 (s, 1H), 7.97 (s, 1H), 7.47 (s, 1H), 6.54 (dd, J = 17.2, 10.2 Hz, 1H), 6.15 (d, J = 17.2 Hz, 1H ), 5.70 (d, J = 10.0 Hz, 1H), 5.12 (dt, J = 12.6, 6.3 Hz, 1H), 3.88-3.79 (m, 1H), 3.12 (s, 4H), 2.48 (s, 4H) , 2.21 (s, 3H), 1.25-1.15 (m, 12H). MS (ESI) m / z: 630 [M + H ⁺ ].

Example 17

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6- (difluoromethoxy) Synthesis of 2- (4-methylpiperazin-1-yl) pyridin-3-yl) acrylamide

Step 1: Synthesis of N- (6-bromo-2-difluoromethoxy-5-nitropyridin-3-yl) acetamide with reference to Example 5.

Step 2: Synthesis of N- (2- (difluoromethoxy) -6- (4-methylpiperazin-1-yl) -5-nitro-pyridin-3-yl) acetamide

At room temperature, N- (6-bromo-2-difluoromethoxy-5-nitropyridin-3-yl) acetamide (1.70 g, 6.03 mmol), acetonitrile (34 ml), and N-methylpiperazine (1.21 g, 12.06 mmol) was heated to 80 ° C. and the reaction was terminated in 3 h. The reaction solution was concentrated and the crude product was subjected to column chromatography to obtain 0.40 g of a brown-yellow oil with a yield of 19%. MS (ESI) m / z: 346.1 [M + H] ⁺ .

Step 3: Synthesis of 2- (difluoromethoxy) -6- (4-methylpiperazin-1-yl) -5-nitro-pyridine-3-amine

At room temperature, add the N- (2- (difluoromethoxy) -6- (4-methylpiperazin-1-yl) -5-nitro-pyridine-3 to the four-necked flask in turn. -Yl) acetamide (0.40 g, 1.16 mmol), 37% hydrochloric acid (0.22 g, 2.32 mmol), the temperature was raised to 60 ° C, and the reaction was terminated in 6 h. The reaction solution was neutralized with 5% NaHCO ₃ to Ph = 8-9, and the solvent was distilled off under reduced pressure at 40 ° C., water (30 ml) was added, and DCM (100 ml x 3) was added for extraction. The organic phases were combined, concentrated, and subjected to crude column chromatography to obtain Brown-yellow oil 0.20 g, yield 24%. MS (ESI) m / z: 304.1 [M + H] ⁺ .

Step 4: 5-Chloro -N ² - (2- (difluoromethoxy) -6- (4-methylpiperazin-l-yl) -5-nitro - pyridin-3-yl) -N ⁴ Synthesis of-(2- (isopropylsulfonyl) pyridin-3-yl) pyrimidine-2,4-diamine

At room temperature, 2- (difluoromethoxy) -6- (4-methylpiperazin-1-yl) -5-nitro-pyridin-3-amine (0.20 g, 0.66 mmol), 2,5-dichloro-N- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidin-4-amine (0.23 g, 0.66 mmol), Pd ₂ (dba) ₃ (0.06 g , 0.06 mmol), Xantphos (0.07 g, 0.13 mmol), K ₃ PO ₄ (0.35 g, 1.65 mmol) and 1,4-dioxane (6 ml), protected by nitrogen, heated to 70 ° C., and the reaction was terminated in 7 h. The reaction solution was cooled to 25-35 ° C, and water (20 ml) was added to quench the reaction, and DCM (10 ml x 2) was added for extraction. The organic phases were combined, concentrated, and subjected to crude column chromatography to obtain a brown-yellow solid 0.30 g with a yield of 27%. MS (ESI) m / z: 614.1 [M + H] ⁺ .

Step 5: N ^2- (5-amino-2- (difluoromethoxy) -6- (4-methylpiperazin-1-yl) pyridin-3-yl) -5-chloro-N ^4- ( Synthesis of 2- (isopropylsulfonyl) pyridin-3-yl) pyrimidine-2,4-diamine

At room temperature, four-necked flask were added 5-Chloro -N ² - (2- (difluoromethoxy) -6- (4-methylpiperazin-l-yl) -5-nitro - pyridine - 3-yl) -N ^4- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidine-2,4-diamine (0.10 g, 0.16 mmol), iron powder (0.045 g, 0.80 mmol), Ammonium chloride (0.044g, 0.80mmol), ethanol (2ml), water (1ml), nitrogen protection, the temperature was raised to 70 ° C, and the reaction was terminated in 1h. The reaction solution was filtered by adding diatomaceous earth, and the filtrate was concentrated. The crude product was subjected to column chromatography to obtain 0.05 g of a yellow solid with a yield of 53%. MS (ESI) m / z: 584.1 [M + H] ⁺ .

Step 6: N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -6- (difluoromethyl Synthesis of oxy) -2- (4-methylpiperazin-1-yl) pyridin-3-yl) acrylamide

At room temperature, N ^2- (5-amino-2- (difluoromethoxy) -6- (4-methylpiperazin-1-yl) pyridin-3-yl) -5 was sequentially added to a four-necked flask. -Chloro-N ^4- (2- (isopropylsulfonyl) pyridin-3-yl) pyrimidine-2,4-diamine (0.05 g, 0.08 mmol), dichloromethane (3 ml), protected by nitrogen, and cooled to Below -30 ° C, a solution of acrylic anhydride (0.011 g, 0.08 mmol) in DCM (1 ml) was added dropwise, and the reaction was terminated in 0.5 h. Was added 5% NaHCO ₃ (3ml) the reaction was quenched by adding DCM (5ml x2). The combined organic phases were concentrated and the crude product by column chromatography to give 0.006 g of off-white solid, yield 11%. MS (ESI) m / z: 638.1 [M + H] ⁺ .

Example 18

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2- (4-methylpiperazine Synthesis of -1-yl) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide

N- (5-((5-chloro-4-((2- (isopropylsulfonyl) pyridin-3-yl) amino) pyrimidin-2-yl) amino) -2- (4-methylpiperazine The method for preparing 1-yl) -6- (2,2,2-trifluoroethoxy) pyridin-3-yl) acrylamide is similar to that in Example 1.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.90 (br, 1H), 9.38 (br, 1H), 9.00 (br, 1H), 8.86 (s, 1H), 8.37 (d, J = 1.4 Hz, 1H), 8.25 (s, 1H), 7.97 (s, 1H), 7.46 (s, 1H), 6.56 (dd, J = 17.0, 10.2Hz, 1H), 6.19 (dd, J = 17.1, 2.0Hz, 1H ), 5.75 (dd, J = 10.1, 2.0 Hz, 1H), 4.90 (q, J = 9.1 Hz, 2H), 3.91-3.81 (m, 1H), 3.26-3.13 (m, 4H), 2.33 (s, 4H), 2.24 (s, 3H), 1.22 (d, J = 6.9Hz, 6H). MS (ESI) m / z: 670.1 [M + H] ⁺ .

Example 19

Test of compounds for kinase inhibitory activity

In this test, the γ- ³³ p-ATP isotope test method was used to test the inhibitory effect of the compound on the kinases EGFR (WT), EGFR (T790M / L858R), and IGF1R, and the compound's half inhibitory concentration of the enzyme's inhibitory activity, IC ₅₀ , was obtained. The drug AZD-9291 is prepared by referring to the method in patent WO2013014448A1.

Basal reaction buffer

20mM Hepes (pH 7.5), 10mM MgCl ₂ , 1mM EGTA, 0.02% Brij35, 0.02mg / ml BSA, 0.1mM Na3VO4, 2mM DTT, 1% DMSO.

2. Compound formulation

The compound was dissolved to a specific concentration with 100% DMSO, and then it was gradient diluted to a different concentration of the test sample (DMSO solution) using an automatic sampling device.

3. Reaction steps

3.1 Dilute the reaction substrate with the basic reaction buffer;

3.2 Add the kinase to the substrate solution and mix gently;

3.3 Use an automatic sample loading system to add 100% DMSO-diluted compounds of different concentrations to the kinase solution and incubate for 20 min at room temperature;

3.4 Add ³³ P-ATP (10 μM, 10 μCi / μl) at room temperature to start the kinase reaction and react for 2 h;

4. Detection

The reaction solution was subjected to ion exchange filtration system to remove unreacted ATP and ADP plasma generated by the reaction, and then measured the ³³ P isotope radiation in the substrate.

5. Data processing

The inhibitory effect of different concentrations of compounds on kinase activity was calculated based on the amount of kinase added to the inhibitor system at different concentrations, and the IC ₅₀ was inhibited by graphpad prism fitting.

The biochemical activity of the compound of the present invention was determined through the above test, and the measured IC ₅₀ value of the enzyme activity is shown in Table 1:

Table 1 Test results of kinase inhibitory activity

Note: "-" in the table means untested.

Conclusion: Compared with the positive control drug AZD-9291, the compound of the present invention has better EGFR (L858R / T790M) mutant kinase inhibitory activity. Examples 1, 2, 3, 5, 7, 9, 12, 13, 14, The compounds of 15, 16, and 18 have better selectivity for EGFR (L858R / T790M) mutant kinase compared to the positive control. The compounds of Examples 7, 8, 9, and 11 have significant relative IGF1R compared to the positive control. Kinase has the advantage of selectivity, and the risk of side effects of elevated blood glucose caused by off-target is smaller.

Example 20

Hcc827 cell proliferation inhibition test

In this experiment, the MTT method was used to test the cell activity of the compound on Hcc827, and the inhibitory concentration IC _{50 of the} compound's inhibitory cell proliferation activity was obtained.

1. Hcc827 cell line was cultured under the condition of RPMI-1640 + 10% FBS. A 96-well cell culture plate was inoculated with 100 μL of a suspension of Hcc827 cells in logarithmic growth phase with a density of 5 × 10 ⁴ / ml. The culture plate was cultured in an incubator for 24 hours to attach the cells (37 ° C, 5% CO ₂ ).

2. Each compound has been dissolved in DMSO to prepare a 10 mM stock solution, which is diluted 400 times the target concentration with DMSO and diluted to 2 times the target concentration with serum-free medium to maintain the DMSO concentration in the drug solution at 0.5. %. In a 96-well plate inoculated with cells, 100 μL / well of each concentration of drug solution was sequentially added. Three replicates were set for each concentration, and a blank control and a negative control were set. The culture was continued at 37 ° C and 5% CO ₂ for 72 hours.

3. Terminate the culture, add 20 μL of MTT solution (5mg / ml) to each well, continue to incubate for 4h at 37 ° C, 5% CO ₂ and discard the medium. Add DMSO 150μL / well, shake at room temperature for 10min, at 490nM wavelength OD values were measured and IC ₅₀ values were calculated by Graphpad Prism 6.0 data processing.

Example 21

NCI-H1975 cell proliferation inhibition test

In this experiment, the MTT method was used to test the cell activity of the compound on NCI-H1975, and the IC ₅₀ value of the half-inhibition concentration of the compound to inhibit cell proliferation activity was obtained.

1. NCI-H1975 cell line was cultured under the condition of RPMI-1640 + 10% FBS, and 100 μL of NCI-H1975 cell suspension in logarithmic growth phase was inoculated in a 96-well cell culture plate with a density of 5X 10 ⁴ / ml, the culture plate was cultured in an incubator for 24 hours to make the cells adhere (37 ° C, 5% CO ₂ ).

2. Discard the RPMI-1640 medium containing 10% FBS in the 96-well cell culture plate and replace it with RPMI-1640 medium containing 0.2% FBS. Each compound has been dissolved in DMSO to prepare a 10 mM stock solution, which is diluted 400 times the target concentration with DMSO and diluted to 2 times the target concentration with serum-free medium. In a 96-well plate inoculated with cells, 100 μL / well of each concentration of drug solution was sequentially added. Three replicates were set for each concentration, and a blank control and a negative control were set. The culture was continued at 37 ° C and 5% CO ₂ for 72 hours.

Terminate the culture, add 20 μL of MTT solution (5mg / ml) to each well, continue to incubate at 37 ° C, 5% CO ₂ for 4 h, discard the medium, add DMSO 150 μL / well, shake at room temperature for 10 min, and measure the OD at a wavelength of 490 nM Value, and calculated IC ₅₀ value by Graphpad Prism data processing.

Example 22

A431 cell proliferation inhibition test

MTT method used in this experiment action of test compounds active in A431 cells and derived compounds inhibit cell proliferation half maximal inhibitory concentration IC ₅₀ activity

1. A431 cell line was cultured under the conditions of DMEM + 10% FBS. A 96-well cell culture plate was inoculated with 100 μL of A431 cell suspension in logarithmic growth phase, with a density of 5 × ^{10 4} / ml. The plate was cultured in an incubator for 24 hours to make the cells adhere (37 ° C, 5% CO ₂ ). .

2. Each compound has been dissolved in DMSO to prepare a 10 mM stock solution, which is diluted 400 times the target concentration with DMSO and diluted to 2 times the target concentration with serum-free medium to maintain the DMSO concentration in the drug solution at 0.5. %. In a 96-well plate inoculated with cells, 100 μL / well of each concentration of drug solution was sequentially added. Three replicates were set for each concentration, and a blank control and a negative control were set. The culture was continued at 37 ° C and 5% CO ₂ for 72 hours.

3. Terminate the culture, add 20 μL of MTT solution (5mg / ml) to each well, continue to incubate for 4h at 37 ° C, 5% CO ₂ and discard the medium. Add DMSO 150μL / well, shake at room temperature for 10min, at 490nM wavelength OD values were measured and IC ₅₀ values were calculated by Graphpad Prism 6.0 data processing.

The biochemical activity of the compound of the present invention was determined through the above tests. The measured IC ₅₀ values of the cells are shown in Table 2:

Table 2 Cell inhibition experiment results

Conclusion: Compared with A431 (EGFR-WT) cells, the compound-positive drug of the present invention has a better inhibitory activity on H1975 (EGFR-T790M / L858R) cells, and the control-positive drug has a significant selectivity for suppressing mutant cell lines.

Claims (10)

1. A compound represented by formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

   

   Wherein R ^{1 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more selected from halogen, hydroxyl, C ₁ -C ₈ alkyl, C ₁ -C ₈ Alkoxy, halogen-substituted C ₁ -C ₈ alkoxy, C ₃ -C ₈ cycloalkyl or C ₃ -C ₈ cycloalkoxy substituted by a substituent;

   R ^{2 is} selected from hydrogen, deuterium, halogen, cyano, nitro, C ₁ -C ₈ alkyl, C ₁ -C ₈ alkoxy, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, trifluoromethyl, difluoromethyl or trifluoromethoxy;

   R ^{3 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of halogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ alkoxy, Halogen-substituted C ₁ -C ₈ alkoxy, C ₃ -C ₈ cycloalkyl, or C ₃ -C ₈ cycloalkoxy;

   X is N or CH;

   R ^{4 is} selected from hydrogen, halogen, cyano, nitro, C ₁ -C ₈ alkyl, halo C ₁ -C ₈ alkyl, -C (O) R ⁵ , -C (O) NR ⁵ R ⁶ , -OR ⁵ , -NR ⁵ R ⁶ , -NR ⁵ C (O) R ⁶ , -NR ⁷ (CH ₂ ) _m NR ⁵ R ⁶ , -NC (O) R ⁷ (CH ₂ ) _m NR ⁵ R ⁶ , -NR ⁷ (CH ₂ ) _m NR ⁵ C (O) R ⁶ , -NR ⁷ (CH ₂ ) _m OR ⁶ , -NC (O) R ⁷ (CH ₂ ) _m OR ⁶ , -O (CH ₂ ) _m NR ⁵ R ⁶ or -O (CH ₂ ) _m NR ⁵ C (O) R ⁶ ;

   R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, wherein C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl is optional Is further substituted with one or more selected from halogen, hydroxy, -NR ⁸ R ⁹ , C ₁ -C ₈ alkyl, C ₁ -C ₈ alkoxy, halogen substituted C ₁ -C ₈ alkoxy, C ₃ -C ₈ cycloalkyl or C ₃ -C ₈ cycloalkoxy substituents;

   Or R ⁵ , R ⁶ and R ⁷ may independently form a 4-10 membered heterocyclic group;

   m is 1, 2, 3 or 4;

   R ⁸ and R ⁹ are each independently selected from hydrogen, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl.
2. The compound of claim 1, wherein:

   R ^{1 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further substituted with one or more substituents selected from halogen or hydroxyl;

   R ^{2 is} selected from hydrogen, deuterium, halogen or methyl;

   R ^{3 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further substituted with one or more substituents selected from halogen;

   X is N or CH;

   R ^{4 is} selected from hydrogen, halogen, cyano, nitro, C ₁ -C ₈ alkyl, -NR ⁵ R ⁶ or -NR ⁷ (CH ₂ ) _m NR ⁵ R ⁶ ;

   R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

   Or R ⁵ , R ⁶ and R ⁷ may independently form a 4-10 membered heterocyclic group;

   m is 1,2,3,4.
3. The compound of claim 1, wherein:

   R ^{1 is} selected from C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl, optionally further substituted by one or more substituents selected from halogen;

   R ^{2 is} selected from hydrogen, deuterium, halogen or methyl;

   R ^{3 is} selected from C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl;

   X is N or CH;

   R ^{4 is} selected from -NR ⁵ R ⁶ or -NR ⁷ (CH ₂ ) _m NR ⁵ R ⁶ ;

   R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl;

   Or R ⁵ , R ⁶ , and R ⁷ may independently form a 6-8 membered heterocyclic group;

   m is 1, 2 or 3.
4. The compound of claim 1, wherein:

   R ^{1 is} selected from methyl, ethyl, difluoromethyl, n-propyl, isopropyl or trifluoroethyl;

   R ^{2 is} selected from hydrogen, deuterium, F, Cl, Br or methyl;

   R ^{3 is} selected from methyl, ethyl, n-propyl, isopropyl or cyclopropyl;

   X is N or CH;

   R ⁴ is -NR ⁷ (CH ₂ ) _m NR ⁵ R ⁶ ;

   R ⁵ , R ⁶ and R ⁷ are each independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl or cyclopropyl;

   Or R ⁵ , R ⁶ , and R ⁷ may independently form a 6-membered heterocyclic group;

   m is 1, 2 or 3.
5. Compound having the following structure or a pharmaceutically acceptable salt thereof:

   

   
6. A pharmaceutical composition comprising the compound according to any one of claims 1 to 5, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
7. The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition is a capsule, powder, tablet, granule, pill, injection, syrup, oral solution, inhalant, ointment, suppository or Patch.
8. The compound according to any one of claims 1 to 5, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 6 is prepared for prevention or treatment alone or partially Application of Epidermal Growth Factor Receptor EGFR Kinase Activity in the Treatment of Diseases
9. A compound according to any one of claims 1 to 5, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 6 in the manufacture of a medicament for preventing or treating cancer application.
10. The application according to claim 9, wherein the cancer is selected from the group consisting of ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, Leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, Acute myeloid leukemia, multiple myeloma, melanoma or mesothelioma.

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