WO2023245611A1

WO2023245611A1 - Small molecule compound with ttk inhibitory activity, preparation method therefor, and use thereof

Info

Publication number: WO2023245611A1
Application number: PCT/CN2022/101072
Authority: WO
Inventors: 母波
Original assignee: 四川大学华西医院
Priority date: 2022-06-24
Filing date: 2022-06-24
Publication date: 2023-12-28

Abstract

The present invention belongs to the field of organic synthesis medicines, and particularly relates to a small molecule compound with TTK inhibitory activity. The general formula of the small molecule compound is shown in formula (I). The small molecule compound with TTK inhibitory activity can be used as a kinase inhibitor targeting TTK, proving that the compound can inhibit the activity of TTK. The compound can be used as an inhibitor for TTK activity and thus be used as a medicament for treating cancers caused by abnormal activation or high expression of TTK, has a good medicinal potential, and provides a new potential choice for clinical medication.

Description

A small molecule compound with TTK inhibitory activity and its preparation method and application

Technical field

The invention relates to the technical field of innovative chemical drugs, and specifically refers to a small molecule compound with TTK inhibitory activity and its preparation method and application.

Background technique

The formation of tumors is the result of abnormal cell proliferation caused by multiple factors such as severe disorder of cell growth regulation and unstable chromosome mitosis. Threonine and tyrosine kinase (TTK), also known as unipolar spindle 1 (Mps1), can ensure the stability of chromosomes. Its abnormal expression can increase chromosome instability, thus leading to the occurrence of tumors. Given the key function of TTK in cell division, it may become a new therapeutic target and biomarker. Except for testis and placenta, it is difficult to detect corresponding expression of TTK in normal human organs, but in triple-negative breast cancer (TNBC), colorectal cancer, ovarian cancer, liver cancer, glioblastoma, thyroid cancer, and pancreatic cancer High expression of TTK is widely found in various human cancers such as , bladder cancer and so on. In fact, TTK is one of 25 genes described as overexpressed in human cancers, and kinase inhibitors targeting TTK are potential anticancer therapeutics. Currently, there are no small molecule kinase inhibitors targeting TTK for clinical application. Based on this, we need to develop a highly efficient targeted TTK kinase inhibitor with a novel backbone structure.

Contents of the invention

The purpose of the present invention is to provide a small molecule compound with novel structure and strong activity that has TTK inhibitory activity.

Another object of the present invention is to provide a method for preparing the above-mentioned small molecule compound.

Another object of the present invention is to provide specific applications of the above small molecule compounds.

The present invention provides a small molecule compound with TTK inhibitory activity, the general formula of which is as follows:

in,

R ₁ is independent methoxy, ethoxy, isopropoxy,

R ₂ is independent

The specific compounds it contains are as follows:

The synthesis route of the above-mentioned small molecule compound with TTK inhibitory activity is as follows:

The raw material is 2-chloro-4-amino-5-methoxypyrimidine.

Its specific preparation method includes the following steps:

(1) 2-chloro-4-amino-5-methoxypyrimidine is demethylated under the action of boron tribromide and methylene chloride as the solvent to prepare intermediate I; the 2-chloro-4 -The molar ratio of amino-5-methoxypyrimidine to boron tribromide is 1:2, the reaction temperature is 0°C, and the reaction time is 18 hours;

(2) Intermediate I is reacted with ethyl 2-bromo-2-methylpropionate under the action of potassium carbonate, using N,N-dimethylformamide as the solvent, to prepare Intermediate II; said Intermediate I : 2-bromo-2-methylpropionic acid ethyl ester: the molar ratio of potassium carbonate is 1:2:2, the reaction temperature is room temperature to 80°C; the reaction time is 18 hours;

(3) Intermediate II reacts with bromocyclopentane under the action of cesium carbonate using N,N-dimethylformamide as a solvent to prepare intermediate III; said intermediate III: bromocyclopentane: The molar ratio of cesium carbonate is 1:2:2; the reaction temperature is room temperature; the reaction time is 18 hours;

(4) Under the catalytic action of Pd(OAc) ₂ , intermediate III undergoes Buchwald-Hartwig coupling reaction with aniline compounds using BINAP as the ligand, cesium carbonate as the base, and 1,4-dioxane as the solvent. , obtain the target compound; the molar ratio of the intermediate III: aniline derivatives: Pd(OAc) ₂ : BINAP: cesium carbonate is 1:1:0.1:0.1:2; the reaction temperature is 100°C; the The reaction time is 16 hours.

The present invention also provides a kinase inhibitor targeting TTK, which is a biopharmaceutically acceptable salt, polymorph, or solvate with the above-mentioned small molecule compound having TTK inhibitory activity as the main active ingredient.

The present invention also provides a drug for treating cancer, which uses the above-mentioned TTK-targeting kinase inhibitor as a main component, and adds a pharmaceutically acceptable, non-toxic, non-inert pharmaceutical carrier and/or an endogenous agent to humans and animals. Prodrugs or pharmaceutical compositions prepared from excipients and auxiliary ingredients.

The pharmaceutical carrier or excipient is one or more solid, semi-solid and liquid diluents, fillers and pharmaceutical product auxiliaries.

The pharmaceutical composition is prepared into various dosage forms using recognized methods in the pharmaceutical and food fields: sprays, aerosols, liquid preparations or solid preparations; the liquid preparations include injections, suspensions, emulsions, solutions or syrups Dosage; the solid preparation includes tablets, capsules, granules or granules.

Cancers that can be treated by the drug include triple-negative breast cancer (TNBC), colorectal cancer, ovarian cancer, liver cancer, glioblastoma, thyroid cancer, pancreatic cancer, and bladder cancer.

The administration route of the drug is oral administration, sublingual administration or mucosal dialysis; the injection includes intravenous injection, intravenous drip, intramuscular injection, intraperitoneal injection or subcutaneous injection.

Compared with the existing technology, the present invention has the following advantages and beneficial effects:

The present invention synthesizes a new type of compound that can be used as a kinase inhibitor targeting TTK, and it is confirmed that the compound can inhibit the activity of TTK. This type of compound can be used as an inhibitor of TTK activity, and can be used as a treatment for abnormal activation of TTK or The use of drugs for cancer caused by high expression has good medicinal potential and provides a new potential choice for clinical medication.

Detailed ways

The present invention will be further described in detail below with reference to the examples, but the implementation of the present invention is not limited thereto. Without departing from the above technical ideas of the present invention, various substitutions and changes can be made based on common technical knowledge and common means in the field. , should be included in the scope of the present invention.

In order to make the purpose, process conditions and advantages of the present invention more clear, the present invention will be further described in detail with reference to the following implementation examples. The specific implementation examples described here are only used to explain the present invention and are not intended to limit the present invention.

Example 1:

Compounds disclosed in this example: 2-[2-methoxy-4-(4-acetylpiperazin-1-yl)anilino]-6,6-dimethyl-8-cyclopentyl-6H-pyrimidine [5,4-b]oxazine-7(8H)-one(I-1)

The specific synthesis route is as follows:

Among them, the raw material is 2-chloro-4-amino-5-methoxypyrimidine

The specific preparation method is as follows:

(1) Weigh the raw material 2-chloro-4-amino-5-methoxypyrimidine (15.95g, 100mmol) into a 500mL round-bottomed flask, then add 200mL of dichloromethane, and then cool to 0 in an ice bath ℃, then boron tribromide (50.10g, 200mmol) was slowly added dropwise. After the dropwise addition, the reaction solution was heated to room temperature and the reaction was continued overnight. After TLC detects that the reaction is complete, the reaction solution is cooled to 0°C, and an appropriate amount of methanol is slowly added to quench the reaction system. Then spin dry to obtain a white powder, which is Intermediate I, which can be directly used in the next step of reaction without purification.

(2) Add Intermediate I (7.25g, 50mmol) and potassium carbonate (13.80g, 100mmol) to a 250mL round-bottomed flask, then add 100mL N,N-dimethylformamide as the solvent and stir at room temperature. Then 2-bromo-2-methylpropionic acid ethyl ester (14.63g, 75mmol) was slowly added dropwise. After the dropwise addition, the temperature of the reaction solution was raised to 80°C, and the reaction was continued overnight. After TLC detects that the reaction is complete, the reaction solution is cooled to room temperature, then filtered and spun dry, and separated and purified by column chromatography to obtain a white powder, which is Intermediate II (7.45g, yield 70%).

The ¹ H NMR data of intermediate II are as follows:

¹ H NMR (400MHz, CDCl ₃ ) δ8.09 (s, 1H), 1.55 (s, 6H); ¹³ C NMR (101MHz, CDCl ₃ ) δ 169.53, 152.21, 149.48, 144.98, 136.28, 80.11, 24.09.MS- ESI(m/z):214.2[M+H] ⁺ .

(3) Add intermediate II (6.39g, 30mmol) and potassium carbonate (8.28g, 60mmol) to a 250mL round-bottomed flask, then add 35mL of N,N-dimethylformamide as the solvent and stir at room temperature. Then bromocyclopentane (6.70 g, 45 mmol) was slowly added dropwise. After the dropwise addition, the temperature of the reaction solution was raised to 80°C, and the reaction was continued overnight. After TLC detects that the reaction is complete, the reaction solution is cooled to room temperature, then filtered and spun dry, and separated and purified by column chromatography to obtain a white powder, which is Intermediate III (5.66 g, yield 67%).

The ¹ H NMR data of intermediate III are as follows:

¹ H NMR (400MHz, CDCl ₃ ) δ8.02(s,1H),5.35–5.26(m,1H),2.07–1.94(m,4H),1.91–1.78(m,2H),1.70–1.53(m ,2H),1.48(s,60H); ¹³ C NMR (101MHz, CDCl ₃ )δ168.91,151.84,150.21,145.09,136.60,79.32,53.51,28.56,25.79,24.09.MS-ESI(m/z):282.4 [M+H] ⁺ .

(4) In a 50mL round-bottomed flask, add intermediate III (0.140g, 0.5mmol) and 2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline (0.125g, 0.5 mmol), palladium acetate (0.011g, 0.05mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (BINAP) (0.062g, 0.1mmol) and cesium carbonate (0.325g, 1.0mmol) , then 10 mL of anhydrous 1,4-dioxane was added as the reaction solvent, and then the reaction was carried out at 100°C for 16 hours under nitrogen protection. After the reaction, the diatomaceous earth was filtered and spin-dried, and then separated and purified by column chromatography to obtain a light yellow powder, which was the target compound I-1 (0.173g, 70%).

The MS (ESI) and ¹ H NMR data of the target compound I-1 are as follows:

MS(ESI):495.8[M+H]+; 1H NMR(400MHz, DMSO-d6)δ7.99(s,1H),7.85(s,1H),7.58(d,J＝8.6Hz,1H), 6.66(s,1H),6.50(d,J＝8.0Hz,1H),5.30–5.08(m,1H),3.79(s,3H),3.63–3.53(m,4H),3.12–3.06(m, 4H),2.05(s,3H),2.05–1.99(m,2H),1.78–1.66(m,4H),1.53–1.45(m,2H),1.40(s,6H).

Example 2:

Compounds disclosed in this example: 2-[2-ethoxy-4-(4-acetylpiperazin-1-yl)anilino]-6,6-dimethyl-8-cyclopentyl-6H-pyrimidine [5,4-b]oxazine-7(8H)-one(I-2)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-ethoxy-4-( N-acetyl-piperazin-1-yl)aniline" to prepare the target compound I-2.

The MS (ESI) and ¹ H NMR data of the target compound I-2 are as follows:

MS (ESI): 509.6[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.01 (s, 1H), 7.80 (s, 1H), 7.62 (d, J = 8.5Hz, 1H ),6.65(s,1H),6.50(d,J＝8.1Hz,1H),5.31–5.07(m,1H),4.06(q,J＝6.9Hz,2H),3.64–3.56(m,4H) ,3.11–3.04(m,4H),2.05(s,3H),2.05–1.99(m,2H),1.83–1.66(m,4H),1.56–1.46(m,2H),1.41(s,6H) ,1.29(t,J＝6.6Hz,3H).

Example 3:

Compounds disclosed in this example: 2-[2-isopropoxy-4-(4-acetylpiperazin-1-yl)anilino]-6,6-dimethyl-8-cyclopentyl-6H- Pyrimidine[5,4-b]oxazin-7(8H)-one(I-3)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-isopropoxy-4- (N-acetyl-piperazin-1-yl)aniline" to prepare the target compound I-3.

The MS (ESI) and ¹ H NMR data of the target compound I-3 are as follows:

MS (ESI): 523.6[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.02 (s, 1H), 7.71 (s, 1H), 7.70 (d, J = 11.0Hz, 1H ),6.67(s,1H),6.52(d,J＝8.4Hz,1H),5.30–5.08(m,1H),4.72–4.48(m,1H),3.64–3.52(m,4H),3.10– 3.03(m,4H),2.04(s,3H),2.04–1.99(m,2H),1.83–1.68(m,4H),1.57–1.46(m,2H),1.41(s,6H),1.24( d,J＝5.7Hz,6H).

Example 4:

Compounds disclosed in this example: 2-[2-(2,2,2-trifluoroethoxy)-4-(4-acetylpiperazin-1-yl)anilino]-6,6-dimethyl -8-Cyclopentyl-6H-pyrimidine[5,4-b]oxazin-7(8H)-one(I-4)

The synthetic route is as in Example 1, replace "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) with "2-(2,2,2- Trifluoroethoxy)-4-(N-acetyl-piperazin-1-yl)aniline" to prepare the target compound I-4.

The MS (ESI) and ¹ H NMR data of the target compound I-4 are as follows:

MS (ESI): 562.6[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.12 (s, 1H), 7.00 (d, J = 8.9Hz, 1H), 6.75 (s, 1H) ),6.61(d,J＝7.7Hz,1H),5.10–5.05(m,1H),4.53–4.51(m,1H),3.62–3.53 (m,4H),3.19–3.13(m,4H), 2.06(s,3H),2.03–1.99(m,2H),1.95–1.83(m,4H),1.62–1.52(s,2H),1.45(s,6H).

Example 5:

Compounds disclosed in this example: 2-[2-methoxy-4-(1-methyl-1H-pyrazole-4yl)anilino]-6,6-dimethyl-8-cyclopentyl-6H -pyrimidine[5,4-b]oxazine-7(8H)-one(I-5)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-methoxy-4-( 1-Methyl-1H-pyrazol-4yl)aniline" to prepare the target compound I-5.

The MS (ESI) and ¹ H NMR data of the target compound I-5 are as follows:

MS(ESI):449.8[M+H] ⁺ ; ¹ H NMR(400MHz, DMSO-d ₆ )δ8.13(s,1H),8.07(s,1H),7.96–7.90(m,2H),7.85 (s,1H),7.21(s,1H),7.12(d,J＝6.7Hz,1H),5.27–5.24(m,1H),3.89(s,3H),3.86(s,3H),2.12– 1.95(m,2H),1.89–1.69(m,4H),1.60–1.49(m,2H),1.42(s,6H).

Example 6:

Compounds disclosed in this example: 2-[2-ethoxy-4-(1-methyl-1H-pyrazole-4yl)anilino]-6,6-dimethyl-8-cyclopentyl-6H -pyrimidine[5,4-b]oxazine-7(8H)-one(I-6)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-ethoxy-4-( 1-Methyl-1H-pyrazol-4yl)aniline" to prepare the target compound I-6.

The MS (ESI) and ¹ H NMR data of the target compound I-6 are as follows:

MS (ESI): 463.4[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.11 (s, 1H), 8.07 (s, 1H), 7.94 (d, J = 8.0Hz, 1H ),7.88(s,1H),7.83(s,1H),7.19(s,1H),7.11(d,J＝8.0Hz,1H),5.32–5.14(m,1H),4.15(q,J＝ 6.7Hz,2H),3.85(s,3H),2.12–1.99(m,2H),1.88–1.73(m,4H),1.60–1.47(m,2H),1.42(s,6H),1.35(t ,J＝6.5Hz,3H).

Example 7:

Compounds disclosed in this example: 2-[2-(2,2,2-trifluoroethoxy)-4-(1-methyl-1H-pyrazole-4yl)anilino]-6,6-di Methyl-8-cyclopentyl-6H-pyrimidine[5,4-b]oxazin-7(8H)-one (I-7)

The synthetic route is as in Example 1, replace "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) with "2-(2,2,2- Trifluoroethoxy)-4-(1-methyl-1H-pyrazol-4yl)aniline" to prepare the target compound I-7.

The MS (ESI) and ¹ H NMR data of the target compound I-7 are as follows:

MS(ESI):517.4[M+H] ⁺ ; ¹ H NMR(400MHz, DMSO-d ₆ )8.19(s,1H),7.91(s,1H),7.37(s,1H),7.23(d,J ＝8.2Hz,1H),7.12(d,J＝8.1Hz,1H),5.15–4.97(m,1H),4.59(dd,J＝17.2,8.4Hz,2H),3.88(s,3H),1.93 –1.83(m,2H),1.59–1.51(m,4H),1.46(s,6H),1.35–1.15(m,2H).

Example 8:

This example discloses a compound: 2-[2-isopropoxy-4-(1-methyl-1H-pyrazole-4yl)anilino]-6,6-dimethyl-8-cyclopentyl- 6H-pyrimidine[5,4-b]oxazin-7(8H)-one(I-8)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-isopropoxy-4- (1-methyl-1H-pyrazol-4yl)aniline" to prepare the target compound I-8.

The MS (ESI) and ¹ H NMR data of the target compound I-8 are as follows:

MS (ESI): 477.4[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.12 (s, 1H), 8.09 (s, 1H), 8.01 (d, J = 8.1Hz, 1H ),7.83(s,1H),7.80(s,1H),7.23(s,1H),7.11(d,J＝7.4Hz,2H),5.41–5.14(m,1H),4.80–4.66(m, 1H),3.85(s,3H),2.08–2.00(m,2H),1.90–1.75(m,4H),1.63–1.51(m,2H),1.43(s,6H),1.31(d,J＝ 5.7Hz,6H).

Example 9:

This example discloses compounds: 2-[2-methoxy-4-(4-hydroxypiperidin-1-yl)anilino]-6,6-dimethyl-8-cyclopentyl-6H-pyrimidine[ 5,4-b]oxazine-7(8H)-one (I-9)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-methoxy-4-( 4-hydroxy-piperidin-1-yl)aniline" to prepare the target compound I-9.

The MS (ESI) and ¹ H NMR data of the target compound I-9 are as follows:

MS (ESI): 468.8[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ7.98 (s, 1H), 7.81 (s, 1H), 7.50 (d, J = 8.6Hz, 1H ),6.60(s,1H),6.46(d,J＝8.1Hz,1H),5.29–5.06(m,1H),4.68(s,1H),3.77(s,3H),3.66–3.55(m, 1H),3.50–3.47(m,2H),2.80(t,J＝10.1Hz,2H),2.07–1.99(m,2H),1.84–1.81(m,2H),1.77–1.63(m,4H) ,1.56–1.46(m,4H),1.40(s,6H).

Example 10:

Compounds disclosed in this example: 2-[2-methoxy-4-(4-hydroxymethylpiperidin-1-yl)anilino]-6,6-dimethyl-8-cyclopentyl-6H- Pyrimidine[5,4-b]oxazin-7(8H)-one(I-10)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-methoxy-4-( 4-hydroxymethyl-piperidin-1-yl)aniline" to prepare the target compound I-10.

The MS (ESI) and ¹ H NMR data of the target compound I-10 are as follows:

MS (ESI): 496.6 [M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ7.99 (s, 1H), 7.76 (s, 1H), 7.54 (d, J = 8.0Hz, 1H ),6.58(s,1H),6.46(d,J＝7.8Hz,1H),5.21–5.17(m,1H),4.51(s,1H),4.03(q,J＝6.8Hz,2H),3.63 (d,J＝11.5Hz,2H),3.30–3.28(m,2H)2.59(t,J＝11.6Hz,2H),2.06–2.00(m,2H),1.79–1.66(m,4H),1.51 –1.47(m,2H),1.40(s,6H),1.31–1.14(m,3H).

Example 11:

Compounds disclosed in this example: 2-[2-isopropoxy-4-(4-hydroxypiperidin-1-yl)anilino]-6,6-dimethyl-8-cyclopentyl-6H-pyrimidine [5,4-b]oxazine-7(8H)-one (I-11)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-isopropoxy-4- (4-hydroxy-piperidin-1-yl)aniline" to prepare the target compound I-11.

The MS (ESI) and ¹ H NMR data of the target compound I-11 are as follows:

MS (ESI): 496.6[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.01 (s, 1H), 7.68 (s, 1H), 7.62 (d, J = 8.3Hz, 1H ),6.60(s,1H),6.48(d,J＝7.3Hz,1H),5.27–5.12(m,1H),4.71(s,1H),4.63–4.53(m,1H),3.60(s, 1H),3.46–3.44(m,2H),2.77(t,J＝9.6Hz,2H),2.11–2.03(m,2H),1.83–1.63(m,6H),1.55–1.45(m,4H) ,1.41(s,6H),1.23(d,J＝5.0Hz,6H).

Example 12:

Compounds disclosed in this example: 2-[2-(2,2,2-trifluoroethoxy)-4-(4-hydroxypiperidin-1-yl)anilino]-6,6-dimethyl- 8-Cyclopentyl-6H-pyrimidine[5,4-b]oxazin-7(8H)-one (I-12)

The synthetic route is as in Example 1, replace "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) with "2-(2,2,2- Trifluoroethoxy)-4-(4-hydroxy-piperidin-1-yl)aniline" to prepare the target compound I-12.

The MS (ESI) and ¹ H NMR data of the target compound I-12 are as follows:

MS (ESI): 536.6[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ7.97 (s, 1H), 7.95 (s, 1H), 7.38 (d, J = 8.6Hz, 2H ),6.72(s,1H),5.24–5.06(m,1H),4.74(s,1H),4.68(dd,J＝17.6,8.8Hz,3H),3.62(s,1H),3.53–3.50( m,2H),2.82(t,J＝10.1Hz,2H),2.04–1.92(m,2H),1.83–1.81(m,2H),1.73–1.57(m,4H),1.51–1.47(m, 2H),1.39(s,6H).

Example 13:

Compounds disclosed in this example: 2-[2-methoxy-4-methanesulfonylanilino]-6,6-dimethyl-8-cyclopentyl-6H-pyrimidine[5,4-b]oxazo Azin-7(8H)-one(I-13)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-methoxy-4-( Replace "N-acetyl-piperazin-1-yl)aniline" with "2-methoxy-4-methanesulfonylaniline" to prepare the target compound I-13.

The MS (ESI) and ¹ H NMR data of the target compound I-13 are as follows:

MS (ESI): 447.6 [M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.42 (d, J = 8.5 Hz, 1H), 8.24 (s, 1H), 8.18 (s, 1H) ),7.51(d,J＝8.5Hz,1H),7.48(s,1H),5.36–5.29(m,1H),3.99(s,3H),3.20(s,3H),2.09–2.00(m, 2H),1.92–1.80(m 4H),1.62–1.58(m,2H),1.44(s,6H).

Example 14:

Compounds disclosed in this example: 2-[2-trifluoromethoxy-4-(N-1-methylpiperidin-4-yl)amidoanilinyl]-6,6-dimethyl-8-cyclo Pentyl-6H-pyrimidine[5,4-b]oxazin-7(8H)-one(I-14)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-trifluoromethoxy-4 -(N-1-methylpiperidin-4-yl)amidoaniline" to prepare the target compound I-14.

The MS (ESI) and ¹ H NMR data of the target compound I-14 are as follows:

MS (ESI): 563.6[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.04 (s, 1H), 8.36 (d, J = 7.7Hz, 1H), 8.12 (s, 1H) ),8.03(d,J＝8.6Hz,1H),7.90(d,J＝8.6Hz,1H),7.86(s,1H),5.27–5.21(m,1H),3.74–3.71(m,1H) ,2.77(d,J＝11.3Hz,2H),2.16(s,3H),2.05–1.98(m,2H),1.93(t,J＝10.9Hz,3H),1.77–1.74(m,6H), 1.64–1.56(m,2H),1.50–1.48(m,2H),1.43(s,6H).

Example 15:

Compounds disclosed in this example: 2-[2-methoxy-4-((1-methylpiperidin-4-yl)oxy)anilinyl]-6,6-dimethyl-8-cyclopentyl -6H-pyrimidine[5,4-b]oxazin-7(8H)-one(I-15)

The synthetic route is as in Example 1, except that "2-methoxy-4-(N-acetyl-piperazin-1-yl)aniline" in step (4) is replaced by "2-methoxy-4-( (1-methylpiperidin-4-yl)oxy)aniline" to prepare the target compound I-15.

The MS (ESI) and ¹ H NMR data of the target compound I-15 are as follows:

MS (ESI): 482.4[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ7.99 (s, 1H), 7.91 (s, 1H), 7.54 (d, J = 8.7Hz, 1H ),6.61(d,J＝1.9Hz,1H),6.52(d,J＝8.6Hz,1H),5.20–5.14(m,1H),4.45–4.18(m,1H),3.76(s,3H) ,2.17(s,3H),2.16–2.14(m,2H),2.04–2.97(m,2H),1.93–1.90(m,2H),1.79–1.54(m,6H),1.45–1.40(m, 2H),1.40(s,6H).

Example 16:

This example is based on the 15 compounds provided in the above examples, and conducts a TTK in vitro kinase activity experiment on them, as follows:

In vitro kinase inhibition analysis was completed using the Kinase Profiler service provided by Eurofins. The experimental method is briefly described as follows: Add the small molecule compound to be tested (0.001-10 μM) or blank solvent, the TTK protein kinase to be tested, and the corresponding polypeptide substrate to the reaction buffer and incubate it, in which the reaction buffer consists of 8mM propylene glycol. Sulfonic acid (MOPS, pH = 7.0), 0.2mM ethylenediaminetetraacetic acid (EDTA), 10mM magnesium acetate and Km concentration of γ- ³³ P-ATP solution. After the entire reaction was carried out at room temperature for 40 min, 3% phosphate solution was added to the reaction buffer to terminate the reaction. Then quantitatively pipette 10 μL of the reaction mixture and drop it on the P30 filter paper, wash it three times with 75mM phosphate, and then wash it once with methanol. After drying the P30 filter paper, add scintillation fluid for scintillation counting. The inhibitory activity of the compound is expressed by the half inhibitory concentration IC _50. The IC ₅₀ value is obtained by fitting the inhibition rate corresponding to each concentration gradient. The results are shown in Table 1. CC-671, a TTK-positive drug, was used as a control group (IC ₅₀ was 0.033 μM).

Table 1 Inhibitory effect of all compounds on TTK activity (IC50)

化合物编号Compound number	EC ₅₀(nM) EC ₅₀ (nM)	化合物编号Compound number	EC ₅₀(nM) EC ₅₀ (nM)
CC-671CC-671	3333	I-8I-8	11951195
I-1I-1	23twenty three	I-9I-9	6262
I-2I-2	3333	I-10I-10	406406
I-3I-3	9797	I-11I-11	225225
I-4I-4	40754075	I-12I-12	4747
I-5I-5	7575	I-13I-13	512512
I-6I-6	358358	I-14I-14	15321532
I-7I-7	842842	I-15I-15	124124

According to the contents of Table 1, it can be seen that each compound has a certain biological activity inhibitory effect. Among them, compound I-1 shows the best biological activity inhibitory effect on TTK, and its IC50 is 0.023 μM.

Claims

A small molecule compound with TTK inhibitory activity, its general formula is as follows:

in,

R 1 is independent methoxy, ethoxy, isopropoxy,

R 2 is independent
A small molecule compound with TTK inhibitory activity according to claim 1, characterized in that when R 1 is a methoxy group, the general structural formula is:

R 2 is one of the following structures:
A small molecule compound with TTK inhibitory activity according to claim 1, characterized in that when R 1 is an ethoxy group, the general structural formula is:

hour,

R 2 is one of the following structures:
A small molecule compound with TTK inhibitory activity according to claim 1, characterized in that when

When R 1 is isopropoxy, the general structural formula is:

hour,

R 2 is one of the following structures:
A small molecule compound with TTK inhibitory activity according to claim 1, characterized in that when R 1 is
When , the general structural formula is:

hour,

R 2 is one of the following structures:
A small molecule compound with TTK inhibitory activity according to claim 1, characterized in that when R 1 is
When , the general structural formula is:

hour,

R 2 is
The preparation method of a small molecule compound with TTK inhibitory activity according to any one of claims 1 to 6, characterized in that it includes the following steps:

(1) 2-chloro-4-amino-5-methoxypyrimidine is demethylated under the action of boron tribromide and methylene chloride as the solvent to prepare intermediate I; the 2-chloro-4 -The molar ratio of amino-5-methoxypyrimidine to boron tribromide is 1:2, the reaction temperature is 0°C, and the reaction time is 18 hours;

(2) Intermediate I is reacted with ethyl 2-bromo-2-methylpropionate under the action of potassium carbonate, using N,N-dimethylformamide as the solvent, to prepare Intermediate II; said Intermediate I : 2-bromo-2-methylpropionic acid ethyl ester: the molar ratio of potassium carbonate is 1:2:2, the reaction temperature is room temperature to 80°C; the reaction time is 18 hours;

(3) Intermediate II reacts with bromocyclopentane under the action of cesium carbonate using N,N-dimethylformamide as a solvent to prepare intermediate III; said intermediate III: bromocyclopentane: The molar ratio of cesium carbonate is 1:2:2; the reaction temperature is room temperature; the reaction time is 18 hours;

(4) Under the catalytic action of Pd(OAc) 2 , intermediate III undergoes Buchwald-Hartwig coupling reaction with aniline compounds using BINAP as the ligand, cesium carbonate as the base, and 1,4-dioxane as the solvent. , obtain the target compound; the molar ratio of the intermediate III: aniline derivatives: Pd(OAc) 2 : BINAP: cesium carbonate is 1:1:0.1:0.1:2; the reaction temperature is 100°C; the The reaction time is 16 hours.
A kinase inhibitor targeting TTK, characterized by a biopharmaceutically acceptable salt using the small molecule compound with TTK inhibitory activity described in any one of claims 1 to 6 as a main active ingredient.
A drug for treating cancer, which is characterized in that it uses the TTK-targeting kinase inhibitor of claim 8 as a main component, and adds a pharmaceutically acceptable, non-toxic, non-inert pharmaceutical carrier to humans and animals, and /or pharmaceutical compositions prepared with excipients and auxiliary ingredients.
A drug for treating cancer according to claim 9, characterized in that the cancers that the drug can treat include triple negative breast cancer (TNBC), colorectal cancer, ovarian cancer, liver cancer, glioblastoma, thyroid cancer Cancer, pancreatic cancer, bladder cancer.