CN113480543B - 2,6, 8-polysubstituted imidazo [1,2-a ] pyrazine and synthetic method and application thereof - Google Patents

Abstract

The present invention discloses a class of 2,6,8-polysubstituted imidazo[1,2-a]pyrazines as described in formula (I) or pharmaceutically acceptable salts thereof. The present invention also discloses that the 2,6,8-polysubstituted imidazo[1,2-a]pyrazine or a pharmaceutically acceptable salt thereof is used as a TYK2 inhibitor in the preparation of a medicine for preventing and treating tumors or inflammatory diseases Applications. The present invention also provides a method for synthesizing the 2,6,8-polysubstituted imidazo[1,2-a]pyrazine or a pharmaceutically acceptable salt thereof.

Description

2,6,8-Poly-substituted imidazo[1,2-a]pyrazine and its synthetic method and application

technical field

The invention belongs to the field of pharmaceutical synthesis and chemical industry, relates to 2,6,8-polysubstituted imidazo[1,2-a]pyrazine, a synthesis method and application thereof, and in particular relates to a 2,6,8- Polysubstituted imidazo[1,2-a]pyrazine compound and preparation method and application thereof.

Background technique

Tyrosine kinase 2 (TYK2) is a member of the JAK family of non-receptor protein tyrosine kinases. death plays an important role. The downstream protein STAT can be recruited by the activated JAK, and then phosphorylated by JAK to form a homologous or heterodimer, enter the nucleus, bind to the promoter of the downstream gene, and regulate the transcription of the downstream gene. The JAK-STAT signaling pathway is closely related to the occurrence and development of tumors and is over-activated in many malignant tumors.

In recent years, more and more studies have shown that TYK2 is closely related to many tumors. In anaplastic large cell lymphoma (ALCL), TYK2 is highly expressed and promotes the growth of ALCL cells through the abnormal activation of the TYK2-STAT1/3-Bcl2 pathway, and the use of TYK2 inhibitors can promote the apoptosis of ALCL cells. TYK2 was found to be generally highly expressed in malignant peripheral nerve sheath tumor (MPNSTs) patient samples, and cell death was significantly increased by lowering TYK2 in murine and human MPNST cells, while TYK2-STAT1/3-Bcl2 was also detected in MPNST Abnormal activation of pathways. In animal studies, it was found that 4T1 breast cancer cells were injected into TYK2(-/-) mice and TYK2(+/+) mice, respectively, and the tumor growth was observed. The ability is obviously stronger than that of TYK2(+/+) mice. Therefore, based on the biological function of TYK2 and the extensive research results in tumors, it can be concluded that TYK2 has an important regulatory role in the occurrence and development of tumors, and TYK2 is a very potential tumor therapy target.

In conclusion, abnormal expression levels of TYK2 are found in samples from patients with various types of tumors, which are closely related to tumorigenesis. Although the reported TYK2 inhibitors have entered the clinical trial stage, there is no TYK2 inhibitor used in tumor therapy. TYK2 is a potential drug target for anti-tumor therapy, and its targeted inhibitors have great potential for the development of anti-tumor drugs. There is an urgent need to develop TYK2 inhibitors for tumor therapy.

SUMMARY OF THE INVENTION

The 2,6,8-polysubstituted imidazo[1,2-a]pyrazine compounds provided by the present invention are TYK2 inhibitors and can be used to treat various tumors, including lymphoma, ovarian cancer, gastric cancer, breast cancer, lung cancer, etc. various malignant tumors.

The 2,6,8-polysubstituted imidazo[1,2-a]pyrazine and its pharmaceutically acceptable salts provided by the present invention are represented by the general structural formula (I):

in:

其中E、Y、A、Q各自独立选自C、N、O、S，在(1)中R₄选自氢基、氨基、甲基氨基、二甲基氨基、羟基、卤原子、氰基、烃基、环烷基、杂原子环烷基、芳基或杂芳环、取代芳基或取代杂芳环、

在(2)中L选自

R₅选自氢基、芳基或杂芳环、取代芳基或取代杂芳环；R ₁ is selected from

Wherein E, Y, A, Q are each independently selected from C, N, O, S, in (1) R ₄ is selected from hydrogen group, amino group, methylamino group, dimethylamino group, hydroxyl group, halogen atom, cyano group , hydrocarbyl, cycloalkyl, heteroatom cycloalkyl, aryl or heteroaromatic ring, substituted aryl or substituted heteroaromatic ring,

In (2) L is selected from

R ₅ is selected from hydrogen, aryl or heteroaromatic ring, substituted aryl or substituted heteroaromatic ring;

当R₂选自(3)中结构时，M、X、Z、T各自独立选自C、 N，R₆选自氢基、氨基、取代氨基、羟基、卤原子、环烷基或含取代基的环烷基、杂原子环烷基或含有取代基的杂原子环烷基、氰基、烃基、芳基或杂芳环、取代芳基或取代杂芳环；R ₂ is selected from cycloalkyl or substituted cycloalkyl, heteroatom cycloalkyl or substituted heteroatom cycloalkyl or

When R ₂ is selected from the structure in (3), M, X, Z, T are independently selected from C, N, and R ₆ is selected from hydrogen, amino, substituted amino, hydroxyl, halogen atom, cycloalkyl or substituted cycloalkyl, heteroatom cycloalkyl or heteroatom cycloalkyl containing substituents, cyano, hydrocarbyl, aryl or heteroaromatic ring, substituted aryl or substituted heteroaromatic ring;

R ₃ is selected from hydrogen atom,

其中E、Y、A、Q各自独立选自C、N，在(1)中R₄选自氨基、氰基，在(2)中L选自

R₅选自

R₂选自环丙烷或

其中M、X、Z、T各自独立选自C、N，R₆选自卤原子、

R₃选自氢原子、

Preferably, R ₁ is selected from

Wherein E, Y, A, Q are each independently selected from C, N, in (1) R ₄ is selected from amino, cyano, in (2) L is selected from

R ₅ is selected from

R ₂ is selected from cyclopropane or

Wherein M, X, Z, T are each independently selected from C, N, R ₆ is selected from halogen atom,

R ₃ is selected from hydrogen atom,

其中E、Y、A、 Q各自独立选自C、N，在(1)中R₄选自氨基、氰基，在(2)中L选自

R₅选自

R₂选自环丙烷或

其中M、X、Z、T各自独立选自C、N，R₆选自氟原子、

R₃选自氢原子、

Further preferably, R ₁ is selected from

Wherein E, Y, A, Q are each independently selected from C, N, in (1) R ₄ is selected from amino, cyano, in (2) L is selected from

R ₅ is selected from

R ₂ is selected from cyclopropane or

Wherein M, X, Z, T are each independently selected from C, N, R ₆ is selected from fluorine atom,

R ₃ is selected from hydrogen atom,

The present invention also provides a pharmaceutical composition, which comprises the compound represented by formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers and/or diluents to prepare Pharmaceutical preparations suitable for clinical use. Wherein, the pharmaceutical composition is formulated into tablets, capsules or liquid preparations and the like. The compounds described in the present invention can be mixed with pharmaceutical carriers or excipients (eg, pharmaceutically acceptable carriers and excipients) according to conventional pharmaceutical formulation techniques to form pharmaceutical formulations. The 2,6,8-polysubstituted imidazo[1,2-a]pyrazine compound can be mixed as an active ingredient in any commonly used oral dosage forms including tablets, capsules and liquids ( such as elixirs and suspensions), which contain coloring agents, flavoring agents, stabilizers and taste-masking substances. For mixed oral dosage forms, the 2,6,8-polysubstituted imidazo[1,2-a]pyrazine compound as an active ingredient can be combined with various common tablet materials (eg starch, calcium carbonate, lactose, sucrose) and dicalcium phosphate) to aid in tableting and encapsulation, the 2,6,8-polysubstituted imidazo[1,2-a]pyrazine compound may be mixed with a pharmaceutically acceptable sterile liquid carrier For example, dissolved or suspended in sterile water, sterile organic solvent, or a mixture of the two. The liquid carrier can be a carrier suitable for injection, such as physiological saline, propylene glycol or aqueous polyethylene glycol solution. In other cases, the micronized active ingredient can also be prepared by dispersing the micronized active ingredient in an aqueous solution of starch or sodium carboxymethyl cellulose or in a suitable oil such as peanut oil. Liquid pharmaceutical preparations (referring to sterile solutions or suspensions) can be administered intravenously, intramuscularly, intraperitoneally or subcutaneously. The pharmaceutical composition may also comprise one or more inorganic or organic, solid or liquid pharmaceutically acceptable carriers or excipients. The term "pharmaceutically acceptable" refers to an additive or composition that is physiologically tolerable and generally does not produce allergic or similar adverse reactions (eg, dizziness, etc.) when administered to an animal such as a mammal (eg, a human). Pharmaceutical carriers and excipients may include, but are not limited to, diluents such as lactose, dextrose, mannose and/or glycerol; lubricants; polyethylene glycols; binders such as magnesium aluminum silicate, starch, gelatin, methyl Cellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone; and, if desired, disintegrating agents such as starch, agar, alginic acid or its salts such as sodium alginate; and/or adsorbents, colorants Agents, Preservatives, Stabilizers, Flavoring and Sweetening Agents.

The present invention also provides the use of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, in the preparation of a TYK2 inhibitor.

The present invention also provides the use of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, in the preparation of a medicine for preventing and treating tumors or inflammatory diseases. The tumors include lymphoma, ovarian cancer, gastric cancer, breast cancer, lung cancer and other malignant tumors.

The compound or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is used for inhibiting the proliferation, growth, infiltration and migration of tumor cells, or promoting the apoptosis of tumor cells.

The present invention also provides a preparation method of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof: obtaining 2-substituted-6,8-dibromopyrazine through ring closure of 2-amino-3,5-dibromopyrazine bromoimidazo[1,2-a]pyrazine, and then undergo a substitution reaction to obtain 2,6-polysubstituted-8-dibromoimidazo[1,2-a]pyrazine, and finally obtain the target compound 2 through coupling reaction ,6,8-Polysubstituted imidazo[1,2-a]pyrazine:

The beneficial effect of the present invention is that the compounds of the present invention are subjected to cell activity assay experiments and TYK2 enzyme activity assay experiments, and the experimental results show that the present compounds have good applications in preventing and treating cancer or inflammation-related diseases.

Detailed ways

The preferred embodiments of the present invention are described in detail below, so that the advantages and features of the present invention can be more easily understood by those skilled in the art, and the protection scope of the present invention can be more clearly defined.

Various aspects and features of the present invention are further described below.

Abbreviations used herein are generally known to those skilled in the art, or may be readily understood from basic knowledge.

The starting materials employed in the preparation of the compounds of the present invention are known, can be prepared according to known methods, or are commercially available.

The present invention also relates to novel intermediates and/or starting materials. The same or similar reaction conditions and novel intermediates as those mentioned in the examples are particularly preferred.

Both intermediates and final products can be worked up and/or purified according to conventional methods including pH adjustment, extraction, filtration, drying, concentration, chromatography, trituration, crystallization, and the like.

Additionally, the compounds of the present invention can also be prepared by various methods known in the art or variations on the methods described herein.

The following examples are intended to illustrate the invention only and not to limit the invention in any way.

Example 1

Preparation of 3-cyano-N-(3-(8-((4-morpholinophenyl)amino)imidazo[1,2-a]pyrazin-6-yl)phenyl)benzamide

Step 1.1: Preparation of 4-(4-nitrophenyl)morpholine (1b)

In a 100 mL one-neck flask, p-fluoronitrobenzene (35.44 mmol) and excess morpholine (205 mmol) were stirred at 80° C. for 5 hours. After the reaction was completed, the reaction mixture was slowly poured into 1000 mL of ice water, and a yellow solid was precipitated, which was filtered and dried to obtain a yellow powdery solid with a yield of 99%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.14 (d, J=9.4 Hz, 2H), 6.84 (d, J=9.4 Hz, 2H), 3.90-3.83 (m, 4H), 3.37 (dd, J =5.9, 4.0Hz, 4H). ¹³ C NMR (101MHz, CDCl ₃ ) δ 155.0, 138.9, 125.9, 112.6, 66.4, 47.1.

Step 1.2: Preparation of 4-morpholinoaniline (1c)

Under argon protection, 4-(4-nitrophenyl)morpholine (19 mmol) was dissolved in 60 mL of ethanol in a two-necked flask, 10% palladium on carbon and hydrazine hydrate were added successively, and the reaction was heated to 75° C. for 8 hours. After the reaction, filter through celite, evaporate the organic solvent under reduced pressure, and separate the target compound by column chromatography with a yield of 95%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 6.68 (d, J=8.8 Hz, 2H), 6.51 (d, J=8.8 Hz, 2H), 4.58 (s, 2H), 3.73-3.65 (m, 4H), 2.88–2.85 (m, 4H); ¹³ CNMR (101 MHz, DMSO-d ₆ ) δ 142.8, 142.8, 118.0, 115.2, 66.8, 51.1.

Step 1.3: Preparation of 6-bromo-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine (1d)

Dissolve 4-morpholine aniline (5.6 mmol) in 60 mL of isopropanol solution, add DIPEA (8.4 mmol) and 6,8-dibromo-imidazo[1,2-a]pyrazine (6.72 mmol) in turn, heat to The reaction was carried out at 85°C for 8 hours. After the reaction, it was cooled to room temperature, the organic solvent was removed under reduced pressure, the residue was dissolved in ethyl acetate, washed with water and saturated brine in turn, and the organic phase was dried with anhydrous sodium sulfate, filtered, and separated by column chromatography on silica gel to obtain White solid, 85% yield. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.81 (s, 1H), 8.18 (d, J=5.7 Hz, 1H), 7.91 (d, J=3.1 Hz, 1H), 7.80 (d, J= 9.3Hz, 2H), 7.60 (d, J=2.4Hz, 1H), 6.92 (d, J=8.3Hz, 2H), 3.73 (d, J=3.1Hz, 4H), 3.06 (dt, J=8.3, 4.6Hz, 4H); ¹³ C NMR (101MHz, DMSO-d ₆ ) δ 147.58, 144.9, 132.9, 132.1, 131.8, 122.0, 121.3, 116.7, 115.7, 111.2, 66.6, 49.4.

Step 1.4: Preparation of 6-(3-Aminophenyl)-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine (1e)

Under the flow of argon, 6-bromo-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine (1 mmol), aminoboronic acid ester (1.2 mmol) were successively added to the two-necked flask. ), PdCl ₂ (dppf) (0.1 mmol), 1M aqueous sodium carbonate solution (1.5 mmol), heated to 100° C. to react overnight. After the reaction was completed, it was cooled to room temperature, filtered through celite, and the organic solvent was evaporated. Ethyl acetate was added to dissolve, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 54%. ¹ H NMR (400MHz, DMSO-d ₆ )δ9.45(s,1H),8.39(s,1H),8.05(d,J=8.6Hz,2H),7.98(s,1H),7.61(s, 1H), 7.22(d, J＝2.2Hz, 1H), 7.12(d, J＝4.7Hz, 2H), 6.99(d, J＝8.6Hz, 2H), 6.59(dt, J＝6.1, 2.7Hz, 1H), 5.42–5.13 (m, 2H), 3.76–3.72 (m, 4H), 3.07 (t, J=4.7Hz, 4H); ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 149.4, 146.8, 145.1, 138.4, 137.6, 133.2, 132.6, 132.6, 129.6, 121.2, 116.7, 116.0, 114.3, 113.8, 111.9, 108.3, 66.6, 49.6.

Step 1.5: 3-Cyano-N-(3-(8-((4-morpholinophenyl)amino)imidazo[1,2-a]pyrazin-6-yl)phenyl)benzamide (1 ) preparation

Under normal temperature, compound 6-(3-aminophenyl)-N-(4-morpholine phenyl) imidazo[1,2-a]pyrazin-8-amine (0.5mmol), 3 -Cyanobenzoic acid (0.7 mmol), and EDCI (0.9 mmol), HOBt (0.9 mmol), DIEA (1.2 mmol), stirred overnight. After the completion of the reaction monitored by TLC, the reaction solution was diluted with ethyl acetate, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain a pure product with a yield of 53%. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.61(s,1H), 9.58(s,1H), 8.70(s,1H), 8.56(s,1H), 8.49(s,1H), 8.34( d, J=8.0Hz, 1H), 8.10 (dd, J=8.3, 5.8Hz, 3H), 8.03 (s, 1H), 7.81–7.72 (m, 2H), 7.67 (d, J=9.9Hz, 2H) ), 7.48(t, J＝7.9Hz, 1H), 7.03(d, J＝8.6Hz, 2H), 3.74(t, J＝4.6Hz, 4H), 3.07(t, J＝4.8Hz, 4H). ¹³ C NMR(101MHz, DMSO-d ₆ )δ164.04,146.79,145.13,139.69,138.12,136.70,136.31,135.51, 133.08,132.96,132.43,131.88,130.31,129.45,121.52,121.48,120.55,118.83, 118.73,117.02 ,116.07,112.04,66.60,49.62.

Example 2

Preparation of 6-(5-aminopyridin-3-yl)-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine (2)

Under argon flow, 6-bromo-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine (1.5mmol) and 5-aminopyridine-3 were successively added to the two-necked flask. -Pinacol borate (1.8 mmol), PdCl ₂ (dppf) (0.15 mmol), 1 M aqueous sodium carbonate solution (3 mmol), heated to 100° C. to react overnight. After the reaction was completed, it was cooled to room temperature, filtered through celite, the organic solvent was removed by rotary evaporation, ethyl acetate was added to dissolve, washed with water and saturated brine successively, and the organic phase was dried with anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 73%. ¹ H NMR (400MHz, DMSO-d ₆ )δ9.51(s,1H), 8.51(s,1H), 8.35(s,1H), 8.07–7.91(m,4H), 7.63(s,1H), 7.48(t, J＝2.3Hz, 1H), 6.99(d, J＝8.5Hz, 2H), 5.51(s, 2H), 3.75(t, J＝4.8Hz, 4H), 3.08(t, J＝4.8 Hz, 4H).; ¹³ C NMR (101MHz, DMSO-d ₆ )δ146.51, 145.00, 135.80, 134.65, 134.45, 133.85, 132.97, 132.46, 132.25, 132.14, 120.97, 116.79, 116.44, 11, 6, 48, 10. .

Example 3

N-(5-(8-((4-morpholinophenyl)amino)imidazo[1,2-a]pyrazin-6-yl)-pyridin-3-yl)-3-(trifluoromethyl) Preparation of benzamide (3)

Under normal temperature, compound 6-(3-aminophenyl)-N-(4-morpholinophenyl) imidazo[1,2-a]pyrazin-8-amine (0.26mmol), 3 - Trifluoromethylbenzoic acid (0.28 mmol), EDCI (0.39 mmol), HOBt (0.39 mmol) and DIEA (0.52 mmol), stirred at room temperature overnight. After the completion of the reaction monitored by TLC, the reaction solution was diluted with ethyl acetate, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 66%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.83(s, 1H), 9.64(s, 1H), 9.04(s, 1H), 8.95(s, 1H), 8.85(d, J=2.4Hz ,1H),8.71(s,1H),8.44-8.32(m,2H),8.11-7.99(m,4H),7.83(t,J＝7.8Hz,1H),7.66(s,1H),7.02( d, J=8.6Hz, 2H), 3.77–3.69 (m, 4H), 3.11– 3.02 (m, 4H). ¹³ C NMR (101 MHz, DMSO-d ₆ ) δ 164.48, 146.58, 145.12, 141.87, 141.08, 135.71 ,135.23,133.55,132.88,132.47,132.41,132.21,131.93,129.89, 125.37,124.59,124.46,124.42,121.29,116.71,115.52,108.97,66.17,49.586

Example 4

Preparation of N-(5-(8-(cyclopropylamino)imidazo[1,2-a]pyrazin-6-yl)-pyridin-3-yl)-3-fluorobenzamide (4)

Step 4.1: Preparation of 6-bromo-N-cyclopropylimidazo[1,2-a]pyrazin-8-amine (4b)

In a 100mL two-necked flask, add 6,8-dibromo-imidazo[1,2-a]pyrazine (9.1mmol) and dissolve it in 20mL isopropanol, followed by adding cyclopropylamine (13.6mmol) and DIPEA (13.6mmol) , heated to 80°C, and reacted for 3 hours. After the reaction was completed, it was cooled to room temperature, and the organic solvent was removed under reduced pressure. After the residue was dissolved in ethyl acetate, a yellow solid was precipitated, which was filtered to obtain pure product with a yield of 92%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.13(d, J=4.3Hz, 1H), 8.04(s, 1H), 7.83(s, 1H), 7.49(s, 1H), 2.91(tt, J=8.1, 3.8Hz, 1H), 0.75–0.63 (m, 4H). ¹³ C NMR (101MHz, DMSO-d ₆ ) δ148.47, 132.03, 131.47, 122.16, 115.81, 109.40, 23.85, 6.16.

Step 4.2: Preparation of 6-(5-Aminopyridin-3-yl)-N-cyclopropylimidazo[1,2-a]pyrazin-8-amine (4c)

Under argon flow, 6-bromo-N-cyclopropylimidazo[1,2-a]pyrazin-8-amine (2.1 mmol) and 5-aminopyridine-3-boronic acid pinacol were added to the two-necked flask in turn. Ester (2.5 mmol), PdCl ₂ (dppf) (0.2 mmol), 1M aqueous sodium carbonate solution (4.2 mmol), heated to 100°C and reacted overnight. After the reaction was completed, it was cooled to room temperature, filtered through celite, and the organic solvent was evaporated. Ethyl acetate was added to dissolve, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 81%. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.37(s, 2H), 8.03-7.84(m, 2H), 7.75 (d, J=4.2Hz, 1H), 7.56-7.49(m, 2H), 5.41(s, 2H), 3.07(tq, J=7.7, 4.0Hz, 1H), 0.77(dh, J=8.1, 3.9Hz, 2H), 0.69(hept, J=4.1Hz, 2H). ¹³ C NMR (101MHz, DMSO-d ₆ )δ148.72,135.72,135.03,134.62,131.87,116.99,115.87,113.79,111.51,109.58,107.07,23.68,6.23.

Step 4.3: N-(5-(8-(Cyclopropylamino)imidazo[1,2-a]pyrazin-6-yl)-pyridin-3-yl)-3-fluorobenzamide (4) preparation

Under normal temperature, add compound 6-(5-aminopyridin-3-yl)-N-cyclopropylimidazo[1,2-a]pyrazin-8-amine (0.4mmol), 3-fluoro Benzoic acid (0.4 mmol), EDCI (0.6 mmol), HOBt (0.6 mmol), and DIPEA (0.6 mmol) were stirred at room temperature overnight. After the completion of the reaction monitored by TLC, the reaction solution was diluted with ethyl acetate, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 28%. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.63(s, 1H), 8.96(d, J=2.0Hz, 1H), 8.91(d, J=2.4Hz, 1H), 8.83(t, J= 2.3Hz, 1H), 8.53(s, 1H), 7.94(s, 1H), 7.85(dd, J=16.8, 7.9Hz, 3H), 7.63(td, J=8.0, 5.8Hz, 1H), 7.54( s, 1H), 7.49 (td, J=8.6, 2.7Hz, 1H), 3.11 (tq, J=7.6, 4.0Hz, 1H), 0.80 (dt, J=6.7, 3.2Hz, 2H), 0.72 (q , J=4.1Hz, 2H). ¹³ C NMR (101MHz, DMSO-d ₆ )δ164.68, 163.16, 160.73, 148.88, 142.14, 141.21, 135.55, 134.05, 133.09, 132.10, 132.08, 130.76, 124.4.68, 124.47 124.02, 118.71, 116.14, 114.75, 114.53, 107.69, 23.73, 6.26.

Example 5

N-(3-(8-Cyclopropylamino)imidazo[1,2-a]pyrazin-6-yl)phenyl)-1H-1,2,4-triazole-3-carboxamide (5 ) preparation

Step 5.1 Preparation of 6-(3-aminophenyl)-N-cyclopropylimidazo[1,2-a]pyrazin-8-amine (5a)

Under argon protection, 6-bromo-N-cyclopropylimidazo[1,2-a]pyrazin-8-amine (4.0 mmol), 3-aminophenylboronic acid pinacol ester ( 4.8 mmol), PdCl ₂ (dppf) (0.4 mmol), 1 M aqueous sodium carbonate solution (8.0 mmol), heated to 100° C. to react overnight. After the reaction was completed, it was cooled to room temperature, filtered through celite, and the organic solvent was evaporated. Ethyl acetate was added to dissolve, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 69%. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.24(s,1H),7.88(s,1H),7.64(d,J=4.2Hz,1H),7.49(s,1H),7.26(t, J=1.9Hz, 1H), 7.09 (dt, J=15.3, 7.7Hz, 2H), 6.56 (dt, J=7.8, 1.6Hz, 1H), 5.12 (s, 2H), 3.08 (tq, J=7.7 , 4.0Hz, 1H), 0.77 (td, J=7.1, 4.6Hz, 2H), 0.68 (q, J=4.0Hz, 2H). ¹³ C NMR (101MHz, DMSO) δ 148.71, 148.39, 138.04, 137.42, 132.03 ,131.71,128.88,115.69,113.66,113.43,111.54,106.46,23.68,6.24.

Step 5.2 N-(3-(8-Cyclopropylamino)imidazo[1,2-a]pyrazin-6-yl)phenyl)-1H-1,2,4-triazole-3-carboxamide (5) Preparation

Under normal temperature, add compound 6-(3-aminophenyl)-N-cyclopropylimidazo[1,2-a]pyrazine-8-amine (0.4mmol), 1H-1,2, 4-Triazole-3-carboxylic acid (0.5 mmol), EDCI (0.7 mmol), HOBt (0.7 mmol), DIPEA (1.02 mmol), stirred at room temperature overnight. After the completion of the reaction monitored by TLC, the reaction solution was diluted with ethyl acetate, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 35%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 14.81(s, 1H), 10.56(s, 1H), 8.59(d, J=34.5Hz, 2H), 8.38(s, 1H), 7.93(s, 1H), 7.82–7.72(m, 3H), 7.52(s, 1H), 7.43(t, J=7.9Hz, 1H), 3.13(tq, J=7.9, 4.1Hz, 1H), 0.80(dq, J ＝6.9, 4.3Hz, 2H), 0.70 (p, J＝4.4Hz, 2H). ¹³ C NMR (101MHz, DMSO) δ 148.57, 146.87, 138.48, 138.01, 136.56, 132.05, 131.91, 131.43, 128.75, 121.44, 120.21 ,118.18,115.94,112.74,106.98,23.71,6.26.

Example 6

Preparation of N-(3-(8-cyclopropylamino)imidazo[1,2-a]pyrazin-6-yl)phenyl)-3-(methylthio)benzamide (6)

Under normal temperature, add compound 6-(3-aminophenyl)-N-cyclopropylimidazo[1,2-a]pyrazine-8-amine (0.6mmol), 3-(methylthio) successively to single-necked flask ) benzoic acid (0.9 mmol), EDCI (1.2 mmol), HOBt (1.2 mmol) and DIPEA (1.6 mmol), and stirred overnight at room temperature. After the completion of the reaction monitored by TLC, the reaction solution was diluted with ethyl acetate, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 72%. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.41(s,1H), 8.49-8.42(m,1H), 8.38(s,1H), 7.97-7.91(m,1H), 7.84(s,1H) ), 7.76(q, J=5.7, 3.9Hz, 4H), 7.54–7.42(m, 4H), 3.13(tt, J=7.5, 3.7Hz, 1H), 2.56(s, 3H), 0.80(dh, J=7.8, 3.5Hz, 2H), 0.71 (q, J=3.9Hz, 2H). ¹³ C NMR (101MHz, DMSO) δ165.18, 148.59, 139.28, 138.74, 137.97, 136.63, 135.67, 132.06, 131.89, 128.99, 128.72, 124.71, 124.19, 121.24, 120.23, 120.19, 118.08, 115.95, 106.93, 23.73, 14.62, 6.28.

Example 7

Preparation of N-(3-(8-cyclopropylamino)imidazo[1,2-a]pyrazin-6-yl)phenyl)-3-(methylthio)benzamide (7)

At 0 °C, N-(3-(8-cyclopropylamino)imidazo[1,2-a]pyrazin-6-yl)phenyl)-3-(methylthio)benzamide (0.4 mmol) was dissolved in dichloromethane and excess m-chloroperoxybenzoic acid (1.6 mmol) was added with stirring. After the reaction, a saturated solution of sodium sulfite was added to separate the layers, the organic phase was dried with anhydrous sodium sulfate, and separated by column analysis to obtain a white solid with a yield of 65%. ¹ H NMR (400MHz, DMSO-d ₆ )δ 10.64(s, 1H), 8.51(t, J=1.8Hz, 1H), 8.45(t, J=2.0Hz, 1H), 8.39(s, 1H), 8.33(dt,J＝7.8,1.4Hz,1H),8.15(dt,J＝7.7,1.4Hz,1H),7.94(d,J＝1.1Hz,1H), 7.85(t,J＝7.8Hz,1H) ),7.80–7.74(m,3H),7.52(d,J=1.1Hz,1H),7.46(t,J=7.9Hz,1H),3.13(tt,J=7.2,3.9Hz,1H),0.80 (td, J=7.1, 4.7 Hz, 2H), 0.73– 0.67 (m, 2H). ¹³ C NMR (101 MHz, DMSO) δ 164.09, 148.61, 141.13, 139.01, 138.07, 136.53, 135.99, 132.75, 132.06, 131.93, 129.86,129.83,128.85,126.20, 121.48,120.35,118.23,115.99,107.00,43.50,23.74,6.29.

Example 8

6-(5-Aminopyridin-3-yl)-N-(4-((1R,4R)-5-isopropyl-2,5-diazabicyclo[2.2.1]heptane-2- Preparation of yl)phenyl)imidazo[1,2-a]pyrazin-8-amine (8)

Under argon flow, 6-bromo-N-(4-((1R,4R)-5-isopropyl-2,5-diazabicyclo[2.2.1]heptane- 2-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine (1.0 mmol), 5-aminopyridine-3-boronic acid pinacol ester (1.2 mmol), PdCl ₂ (dppf) (0.1 mmol), 1M aqueous sodium carbonate solution (2.0 mmol), heated to 100 °C and reacted overnight. After the reaction was completed, it was cooled to room temperature, filtered through celite, and the organic solvent was evaporated. Ethyl acetate was added to dissolve, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 30%. ¹ HNMR (400 MHz, DMSO-d ₆ )δ9.43(s,1H), 8.47(s,1H), 8.34(s,1H), 7.98–7.92(m,4H), 7.62(s,1H), 7.45(d, J=2.7Hz, 1H), 6.68(d, J=8.5Hz, 2H), 5.48(s, 2H), 4.40(s, 1H), 3.21(s, 5H), 2.89–2.59(m , 2H), 1.97(s, 1H), 1.12–1.01(m, 6H). ¹³ C NMR(101 MHz, DMSO) δ145.05, 144.73, 142.50, 142.32, 135.96, 134.81, 134.68, 133.01, 132.21, 132.17, 121.61 ,116.71,116.45,112.85,108.16,59.81,58.55,57.98,56.40,55.02,20.82,14.13.

Example 9

Preparation of 6-(3-aminophenyl)-N-(5-fluoropyrimidin-2-yl)imidazo[1,2-a]pyrazin-8-amine (9)

Step 9.1 Preparation of 6-bromo-N-(5-fluoropyrimidin-2-yl)imidazo[1,2-a]pyrazin-8-amine (9b)

Under argon flow, 6,8-dibromo-imidazo[1,2-a]pyrazine (3.6 mmol), 2-amino-5-fluoro-pyridine (5.4 mmol), Pd ₂ ( dba) ₃ (0.36 mmol), Xantphos (0.72 mmol) and cesium carbonate (7.2 mmol), heated to 100°C and reacted overnight. After the reaction was completed, it was cooled to room temperature, filtered through celite, and the organic solvent was evaporated. Ethyl acetate was added to dissolve, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 45%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.33 (s, 1H), 8.41 (s, 1H), 8.38 (d, J=3.1 Hz, 1H), 8.33 (dd, J=9.2, 4.1 Hz, 1H), 8.01(d, J=1.1Hz, 1H), 7.87(td, J=8.7, 3.1Hz, 1H), 7.68(d, J=1.1Hz, 1H). ¹³ C NMR(101MHz, DMSO)δ147 .95,143.08,135.76, 135.51,133.42,131.35,125.58,125.38,119.71,116.84,114.32,113.32.

Step 9.2 Preparation of 6-(3-aminophenyl)-N-(5-fluoropyrimidin-2-yl)imidazo[1,2-a]pyrazin-8-amine (9)

Under argon flow, 6-bromo-N-(5-fluoropyrimidin-2-yl)imidazo[1,2-a]pyrazin-8-amine (2.0 mmol) and 3-aminobenzene were successively added to the two-necked flask. Pinacol borate (2.4 mmol), PdCl ₂ (dppf) (0.2 mmol), and 1M aqueous sodium carbonate solution (4.0 mmol) were heated to 100° C. to react overnight. After the reaction was completed, it was cooled to room temperature, filtered through celite, and the organic solvent was evaporated. Ethyl acetate was added to dissolve, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 45%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.96 (s, 1H), 8.74 (dd, J=9.2, 4.0 Hz, 1H), 8.58 (s, 1H), 8.37 (d, J=3.1 Hz, 1H), 8.06(d, J＝1.1Hz, 1H), 7.89(td, J＝8.7, 3.1Hz, 1H), 7.68(d, J＝1.1Hz, 1H), 7.24(t, J＝1.9Hz, 1H), 7.16–7.09(m, 2H), 6.61(dt, J=7.5, 1.9Hz, 1H), 5.27(s, 2H). ¹³ C NMR (101MHz, DMSO) δ 156.42, 153.97, 149.05, 148.66, 143.08, 137.27, 136.70, 135.63, 135.38, 133.07, 131.69, 129.31, 125.61, 125.42, 116.85, 113.97, 113.74, 113.70, 113.21, 111.42, 109.89.

Example 10

Preparation of 3-cyano-N-(3-(8-((5-fluoropyrimidin-2-yl)amino)imidazo[1,2-a]pyrazin-6-yl)phenyl)benzamide

At room temperature, the compound 6-(3-aminophenyl)-N-(5-fluoropyrimidin-2-yl)imidazo[1,2-a]pyrazin-8-amine (0.5mmol) was added successively to the single-necked flask , 3-cyanobenzoic acid (0.7 mmol), and EDCI (0.9 mmol), HOBt (0.9 mmol), DIPEA (1.2 mmol), and stirred overnight at room temperature. After the completion of the reaction monitored by TLC, the reaction solution was diluted with ethyl acetate, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 77%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.54 (s, 1H), 8.96 (s, 1H), 8.86 (dd, J=9.2, 4.1 Hz, 1H), 8.73 (s, 1H), 8.68 (s,1H),8.47(s,1H),8.33(dd,J=9.5,5.4Hz,2H),8.08(d,J=8.9Hz,2H),7.83(ddd,J=21.8,10.9,5.5 Hz, 2H), 7.67 (td, J=11.3, 9.2, 4.8 Hz, 3H), 7.46 (t, J=7.9 Hz, 1H). ¹³ C NMR (101 MHz, DMSO) δ 163.78, 156.46, 154.01, 148.55, 143.15 ,139.40,137.13,135.96,135.70,135.56,135.31,135.02, 133.14,132.58,131.70,131.35,129.87,129.05,125.54,125.35,120.91,119.96, 118.39,117.84,117.04,114.08,111.55,110.31.

Example 11

Preparation of 6-(3-cyanophenyl)-N-methyl-8-((4-morpholinophenyl)amino)imidazo[1,2-a]pyrazine-2-benzamide (11)

Step 11.1 Preparation of ethyl 6,8-dibromoimidazo[1,2-a]pyrazine-2-carboxylate (11b)

In a 100 mL single-necked flask, 2-amino-3,5-dibromopyrazine (20 mmol) and ethyl 3-bromopyruvate (24 mmol) were dissolved in 40 mL of ethylene glycol dimethyl ether, and stirred at 100° C. overnight. After the reaction was completed, ether was added to the reaction mixture, filtered and dried to obtain a brown solid with a yield of 47%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.31 (dd, J=7.2, 1.7 Hz, 2H), 4.48 (qd, J=7.1, 1.6 Hz, 2H), 1.43 (td, J=7.1, 1.7 Hz, 3H). ¹³ C NMR (101MHz, CDCl ₃ ) δ 161.81, 139.73, 138.31, 135.93, 121.68, 119.55, 119.32, 62.14, 14.46.

Step 11.2: Preparation of ethyl 6-bromo-8-((4-morpholinophenyl)amino)imidazo[1,2-a]pyrazine-2-carboxylate (11c)

4-Mormorpholinaniline (7.2 mmol) was dissolved in 30 mL of acetonitrile solution, DIPEA (16.2 mmol) and ethyl 6,8-dibromoimidazo[1,2-a]pyrazine-2-carboxylate (6.0 mmol) were added successively ), heated to 85°C and reacted overnight. After the reaction, it was cooled to room temperature, the organic solvent was removed under reduced pressure, the residue was dissolved in ethyl acetate, washed with water and saturated brine in turn, and the organic phase was dried with anhydrous sodium sulfate, filtered, and separated by column chromatography on silica gel to obtain Yellow solid, 39% yield. ¹ H NMR (400MHz, CDCl ₃ ) δ 8.13(s, 1H), 8.00(s, 1H), 7.70(d, J=8.6Hz, 2H), 7.60(s, 1H), 6.92(d, J= 8.5Hz, 2H), 4.44 (q, J=7.1Hz, 2H), 3.85 (t, J=4.5Hz, 5H), 3.12 (t, J=4.6Hz, 4H), 1.41 (d, J=14.1Hz) , 3H). ¹³ C NMR (101MHz, CDCl ₃ )δ162.31, 148.02, 144.66, 136.04, 132.43, 130.76, 123.83, 121.06, 118.96, 116.32, 110.01, 66.94, 61.57, 49.68, 14.47

Step 11.3: Preparation of 6-bromo-N-methyl-8-((4-morpholinophenyl)amino)imidazo[1,2-a]pyrazine-2-carboxamide (11d)

Dissolve ethyl 6-bromo-8-((4-morpholinophenyl)amino)imidazo[1,2-a]pyrazine-2-carboxylate (2.35mmol) in 10mL of tetrahydrofuran solution, add 12mL of 30% The aqueous methylamine solution was stirred at room temperature overnight. After the reaction was completed, it was filtered to obtain a pale yellow solid with a yield of 85%. ¹ H NMR (400MHz, CDCl ₃ ) δ 8.01(s, 1H), 7.79(s, 1H), 7.74(d, J=8.5Hz, 2H), 7.67(s, 1H), 7.23(d, J= 5.6Hz, 1H), 7.00(s, 2H), 3.90(s, 4H), 3.17(t, J=4.8Hz, 4H), 3.05(d, J=5.0Hz, 3H). ¹³ C NMR(101MHz, CDCl3)δ162.84,149.54,144.59,137.06,133.61,130.59, 123.82,121.06,119.08,116.31,110.76,77.48,77.16,76.84,66.92,61.67,26.07.

Step 11.4: 6-(3-Cyanophenyl)-N-methyl-8-((4-morpholinophenyl)amino)imidazo[1,2-a]pyrazine-2-benzamide (11 ) preparation

Under argon flow, 6-bromo-N-methyl-8-((4-morpholinophenyl)amino)imidazo[1,2-a]pyrazine-2-carboxamide (0.23 mmol), 3-cyanophenylboronic acid pinacol ester (0.27 mmol), PdCl ₂ (dppf) (0.02 mmol), 1M aqueous sodium carbonate solution (0.46 mmol), heated to 100° C. to react overnight. After the reaction was completed, it was cooled to room temperature, filtered through celite, and the organic solvent was evaporated. Ethyl acetate was added to dissolve, washed with water and saturated brine successively, and the organic phase was dried over anhydrous sodium sulfate. It was separated by column chromatography on silica gel to obtain pure product with a yield of 42%. ¹ H NMR (400MHz, DMSO-d ₆ )δ9.41(s, 1H), 8.72(s, 1H), 8.35(d, J=1.8Hz, 1H), 8.32-8.28(m, 2H), 8.14( q,J=4.8Hz,1H),7.92-7.88(m,2H),7.85(dt,J=7.7,1.4Hz,1H),7.71(t,J=7.8Hz,1H),7.02-6.97(m , 2H), 3.77–3.73 (m, 4H), 3.12–3.07 (m, 4H), 2.86 (d, J=4.8Hz, 3H). ¹³ C NMR (101MHz, DMSO-d ₆ )δ161.85, 147.01, 145.19 ,139.32,138.14,135.12, 131.70,131.62,131.55,130.22,130.13,128.96,121.36,118.83,117.14,115.38, 111.88,109.47,66.17,48.95,25.60

Example 12

biological assay

TYK2 enzyme activity assay

A 384-well microplate TR-FRET (time-resolved-fluorescence energy transfer) endpoint assay was used for TYK2 kinase activity assay. The same assay was used for _IC50 determination of small molecule inhibitors. Typically, kinase reactions are performed in a 25 μL volume of reaction solution containing: 5 μL compound (in 10% DMSO), 20 μL in assay buffer (25 mM HEPES, pH 7.5, 0.001% Brij-35, 0.01% A mixture of TYK2, polypeptide and ATP in Triton, 0.5 mM EGTA). All reactions were performed in 384-well white flat-bottom optical density plates (Perkin Elmer) for 60 min at room temperature, followed by the addition of 25 μL of stop reaction (100 mM HEPES, pH 7.5, 0.015% Brij-35, 0.2% Coating Reagent #3, 50 mM EDTA) . Read on an Evision Multilabel reader (Perkin Elmer, Envision 2102-0010).

Table 1

Example number Inhibition against TYK2 at 1μM(%) 1 76%(IC₅₀=0.2μM) 2 55% 3 3% 4 10% 5 8% 6 6% 7 3%

Cell viability assay

The cells in the logarithmic growth phase were seeded in a 96-well plate at a density of 3000 cells/100 μL per well. After the cells adhered, 100 μL of different concentrations of the compounds to be tested were added to obtain 6-8 concentration gradients. Five parallel wells were set in each group, and a control group was set. After the compound and tumor cells were incubated for 72 hours, 10 μL of CCK-8 solution was added to each well. After 1-2 hours of incubation in the cell incubator, measure the absorbance (OD value) of each well with an enzyme-linked immunosorbent assay, and calculate the inhibition rate: inhibition rate (IR%) = (1-TOD/COD) x 100%, TOD : mean OD value of drug group; COD: mean OD value of solvent control group.

Table 2

The above embodiments are only intended to illustrate the technical concept and characteristics of the present invention, and the purpose is to allow those who are familiar with the technology to understand the content of the present invention and implement it, and cannot limit the scope of protection of the present invention. Equivalent changes or modifications made should all be included within the protection scope of the present invention.

Claims (6)

1. A compound or a pharmaceutically acceptable salt thereof, characterized by the following:

。

2. a pharmaceutical composition comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof together with one or more pharmaceutically acceptable carriers and/or diluents, in a clinically acceptable pharmaceutical formulation.

3. The pharmaceutical composition of claim 2, wherein the pharmaceutical formulation is selected from the group consisting of a tablet, a capsule, and a liquid formulation.

4. Use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 2, in the preparation of an inhibitor of TYK 2.

5. The use of the compound or pharmaceutically acceptable salt thereof according to claim 1, or the pharmaceutical composition according to claim 2 for the preparation of a medicament for the prevention and treatment of neoplastic or inflammatory diseases.

6. The use of claim 5, wherein said compound or pharmaceutically acceptable salt thereof, or said pharmaceutical composition is used to inhibit proliferation, growth, infiltration and migration of tumor cells, or to promote apoptosis of tumor cells.